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Hybridization: Importance, Techniques and

Consequences; Recent Trends in Agriculture

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 Recent Trends in
Agriculture

Editors
Dr. Srishti Thakur, Rhitisha Sood, Rukoo Chawla, Kiran C and Niharika
Sheoran

Integrated Publications TM
New Delhi
Published By: Integrated Publications TM

Integrated Publications
H. No. - 3 Pocket - H34, Sector - 3,
Rohini, Delhi-110085, India
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Editors: Dr. Srishti Thakur, Rhitisha Sood, Rukoo Chawla, Kiran C and
Niharika Sheoran

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© Integrated Publications
Edition: 1st
Publication Year: 2023
Pages: 321
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 Content

S. No. Chapters Page No.

1. Nano Urea - A Bliss for Agriculture or Not 01

(Kannoj, Manish Tomar, Ruchika Choudhary and Ritesh Patidar)

2. Disease Resistance in Insects 19

(Jasra Bano, Qadrul Nissa and Jamal Ahmad)

Nuclear Polyhedrosis Virus (NPV): Role and

3. Importance for Crop Pest Management 31
(Selvam K, Shiva N and Manikandan P)

4. Post-harvest Technology for Fruits 53

(Aakash, Mrinali Sood, Garima and Deepak Kumari)

5 Nutrient Use Efficiency 71

(Abhishek Raj Ranjan and Anurag Singh Suman)

6. Natural Farming: An Eco-agriculture 91

(Komal Sharma, Minam Gamoh and Nisha Devi)

Rhizosphere: Role and Importance for Soil Health

7. Management 105
(Varun Parmar, Naveen Datt and Rushali Katoch)

CRISPR - Genome Editing Tools for Genetic

8. Improvement in Agriculture 123
(Prachi Mahla, Sheenam Bhateja and Himansuman)

Various Methods of Testing Hypotheses and

9. Measure of Dispersion in Statistics 143
(Shiv Kumar Rana)

10. Emerging Challenges of INM in Crop Management 155

(Saloni Yadav, Amit Kumar, Asha and Mohit Sharma)

Grafting: Types, Advantages and Disadvantages for

11. the Crop Improvement 167
(BC Akhilraj and J Suresh)

Hybridization: Importance, Techniques and

12. Consequences 185
(Manika Goswami, Sarita, Mohammad Ashiq Kuchay and Davinder)

Role and Importance of ICT in Agricultural

13. Extension 199
(Shruti Samadder and Deepak Rao)
 Nuclear Polyhedral Virus (NPV): Role and
14. Importance for Crop Pest Management 215
(Mary Lisha J, Saai Vignesh B and Balaji K)

15. Canopy Management 227

(Megha Patidar and Parshant Bakshi)

16. Breeding for Improved Quality in Vegetable Crops 241

(Sheetal Rana)

Scheduling of Irrigation for Various Major Crops

17. with Importance and Limitation in Agriculture 255
(Kalyani MSR, Archana B, Srinivas Y and Vijaykumar P)

18. Precision and Regenerative Agriculture 275

(Asha, Diksha, Shabnam and Priyanka Sanwal)

19. Future of Nanotechnology in Agriculture 291

(Sonal Kumari and Ved Prakash)
 Chapter - 1
Nano Urea - A Bliss for Agriculture or Not

Authors
Kannoj
Department of Agronomy, CCS Haryana Agricultural
University, Hisar, Haryana, India
Manish Tomar
Department of Agronomy, CCS Haryana Agricultural
University, Hisar, Haryana, India
Ruchika Choudhary
Department of Agronomy, Rajasthan College of Agriculture,
MPUAT, Udaipur, Rajasthan, India
Ritesh Patidar
Department of Agronomy, Rajasthan College of Agriculture,
MPUAT, Udaipur, Rajasthan, India

Page | 1
Page | 2
 Chapter - 1
Nano Urea - A Bliss for Agriculture or Not
Kannoj, Manish Tomar, Ruchika Choudhary and Ritesh Patidar

Introduction
World agricultural cropping systems use a considerable quantity of
fertilizers, insecticides and herbicides to increase productivity per unit area,
but at higher than optimal levels of these chemicals and fertilizers. Mittal et
al. (2020) estimated that global fertilizer usage will rise by 263 mt in 2050,
owing to the increased need of fertilizers for higher yield. Further, according
to Fatima et al. (2021), fertilizer application grew 13 times between 1950 and
2020, from 15 to 194 million tonnes. This excessive use causes a variety of
issues, including environmental pollution (soil, water, and air pollution), low
input use efficiency, decreased food quality, resistance development in various
weeds, diseases, and insects, lower income from production, soil degradation,
micronutrient deficiency in the soil, adverse effect on other beneficial living
organisms presents above and below the soil surface, and so on. It has been
claimed that leaching, mineralization, and bioconversion waste 50-70 per cent
of chemical inputs (Bollag et al., 1992). Chemical fertilizer residues harmed
human health and destabilised ecosystem sublevels (such as soil microbial
flora, parasites, and the marine environment) through runoff and
eutrophication (Conley et al., 2009). As a result, there is a need to transform
traditional agricultural operations into creative approaches for sustainable
agriculture using innovative techniques such as nanotechnology.
Nitrogen plays an essential role in the growth and development of plants
required for chlorophyll, synthesis of enzymes and proteins, metabolites and
nucleic acids. India is the second-largest consumer and the third-largest
producer of urea in the world. In India, the total consumption and production
are 191.01 and 136.85 lakh tonnes, respectively, which was not fulfilling the
farmer’s demand, so the government of India imported 51.91 lakh tonnes of
nitrogen fertilizer (Pocket book of agricultural statistics, 2020). Urea
contributes about 82 per cent of the total fertilizer consumption in India and
about 55 per cent of the total fertilizer nitrogen consumed worldwide. It is the
most extensively used nitrogen source due to several characteristics, including

Page | 3
its high solubility in water, non-polarity, rapid leaf absorption and low
phytotoxicity (Bondada et al., 2001). Around 30-40 per cent of urea nitrogen
is consumed by plants, with the remainder being squandered owing to rapid
chemical change as a result of leaching, volatilization, denitrification, and
runoff, resulting in low utilisation efficiency. Nano urea, on the other hand,
has a high nitrogen utilisation efficiency and is also environmentally
beneficial. This fertilizer is generally known as "smart fertilizer" since it
minimises nitrous oxide emissions, which are primarily responsible for
polluting soil, air, and water bodies. It also contributes to global warming
reduction. These properties make it a promising alternative to conventional
urea.
Nanotechnology has promising potential for sustainable agriculture and
is prophesied to transform traditional agriculture techniques into precision
farming. Norio Taniguchi coined the term "nanotechnology" in 1974. The use
of nanoformulation, nanoencapsulation, and functionalized nanomaterials in
next-generation fertilizers and pesticides allows for site-specific and regulated
distribution of active chemicals (fertilizers and protectants) to plants, as well
as a reduction in excess run-off (Nair et al., 2010; Ghormade et al., 2011; Khot
et al., 2012). It is hypothesized that nanotools perform competent delivery and
their controlled release capacity of agriculture inputs could be helpful in
maintaining ecological balance and providing sustainable solutions to climate
change and environmental pollution (Solanki et al., 2015)
Nano fertilizers can penetrate the plant tissue more easily than other
traditional forms of fertilizers because they have a large surface area relative
to their size. Nano fertilizers, a new generation of fertilizers, will raise nutrient
use efficiency from 30 per cent to as high as 80 per cent. The effectiveness of
the use of nutrients (NUE) is a crucial metric for assessing the development
of better nutrient management. Many emerging nations rely heavily on
agriculture as their economic engine. The primary rationale for using fertilizer
in agriculture is to provide the soil with the complete macro and
micronutrients that it generally lacks. Fertilizer contributes between 35 and 40
per cent of agricultural production.
Additionally, the site-specific and regulated release of agricultural inputs'
active ingredient lowers the amount and cost of fertilizer borne by farmers
(Mukhopadhyay, 2014). Further, eliminating chemical fertilizers would be
beneficial in increasing soil health and preserving the geobiological soil cycle,
which would boost crop production's, food and nutrition quality. So, there is a
need to upgrade conventional agricultural operations into creative practises by
using advanced techniques like nanotechnology for sustainable agriculture.

Page | 4
 Applying cutting-edge nanotechnology to agriculture (including the
creation of fertilizers) is one of the potential strategies in sustainable
agriculture that might help to feed the world's fast-expanding population. This
has led to looking for environmentally friendly or sensible fertilizers, chiefly
those with high nutrient-use potency. Applied science is rising with a
promising alternative i.e., Nano fertilizers.
Nano-fertilizer
The term "nano" is derived from the Greek word for "dwarf," or one
billionth of a metre and particles which are smaller than 100 nm are termed
nanoparticles (Thakkar et al., 2010). Nano fertilizers are chemically
synthesized or amended forms of traditional fertilizers, fertilizer bulk
materials, or extracts from various vegetative or reproductive parts of the plant
using different chemical, physical, mechanical, or biological methods and are
used to improve soil fertility, productivity, and agricultural produce quality
(Meena et al., 2017).
These are nanomaterial or nano-scale fertilizer formulations that are
accountable for providing one or more types of nutrients to growing plants,
hence stimulating their growth and improving production (Liu and Lal, 2015).
Nano-fertilizers are nanoparticles that employ nanotechnology to boost
nutrients and increase their use efficiency (Khalid et al., 2022). Nanoparticles
are more efficient than bulk components because of their tiny size and high
surface-to-volume ratio. Nano fertilizers are projected to significantly boost
crop growth and yields, improve fertilizer efficiency, reduce nutrient losses,
and limit negative environmental consequences compared to traditional
fertilizers.
Nano fertilizer versus conventional fertilizers
Presently, Nanotechnology in agriculture focuses on target farming which
employs nanosized particles with unique properties to boost crop production
while minimising adverse environmental impact (Batsmanova et al., 2013;
Scott and Chen, 2013). Nano fertilizers are more advantageous than traditional
fertilizers since they improve soil fertility, crop productivity and crop quality
metrics. They are non-toxic and less hazardous to the environment, humans
and reduce costs while increasing profits. Nano-fertilizers have shown
superior transport capabilities by plant cells and tissues with regulated
mobility as compared to traditional water-soluble fertilizers (Al-Mamun et al.,
2021). Furthermore, traditional fertilizers are not useful in other respects, such
as organic matter gathered from diverse plants and animals being dissimilar.
It cannot supply the same amount of nutrients and organic matter as fertilizer.
(Toksha et al., 2021).

Page | 5
Nano fertilizers are formulated to meet the nutritional requirements of
intended crops. The nano form of traditional Agri-inputs allows for the site-
specific and regulated release of active substances, reducing excess run-off
and preventing eutrophication and residual contamination. Nanoparticles
improve nutrient usage efficiency and reduce environmental protection costs
by reducing salt build-up in the soil, as they are needed in less quantity. All of
these will lead to an increase in crop nutritional content and seasoning quality.
Accelerate growth and development of plants by combating illnesses and
increasing plant stability through anti-bending and profound rooting of crops.
As a result, these nutrient formulations may serve as alternatives to traditional
fertilizers, assuring extended and regulated nutrient release. However, there
may be some important restrictions to the extensive use of nano fertilizers in
agriculture that must be considered. The toxicity/biocompatibility of nano
fertilizers must be investigated.

Fig: Effect of nano fertilizers on plant growth

Categories of nano-fertilizers
1) Nano-scale fertilizer (nanoparticles that contain nutrients).
2) Nano-scale additives (traditional fertilizers with nano-scale
additives).
3) Nano-scale coating (traditional fertilizers coated or loaded with
nanoparticles).
Nanomaterial coverings (such as a nanomembrane) might limit nutrient
release, or a porous nano fertilizer could have a network of channels that slow
nutrient solubility. Another exciting use of applied science is the improvement
of plant root health by encapsulating helpful bacteria. These might include a

Page | 6
variety of microbes or fungi that increase the availability of chemical
elements, phosphorus, and potassium in the root zone.
IFFCO nano urea
On May 31, 2021, the Indian Farmers Fertilizer Cooperative Limited
(IFFCO) launched the world's first nano-liquid urea (Nano urea). It has been
developed indigenously at Nano Biotechnology Research Centre, Kalol,
Gujrat, in line with Atmanirbhar Bharat and Atmanirbhar Krishi. It will be
used to supplement plant nutrition in place of conventional urea. On World
Environment Day, IFFCO delivered its first shipment of Nano Urea Liquid to
Uttar Pradesh for usage by farmers. Nano urea initiates a revolutionary change
in agriculture by increasing production, lowering pollution and saving money
on government subsidies. Nano urea is the liquid version of solid urea, and a
500 ml bottle contains 40,000 ppm nitrogen, providing nitrogen nourishment
comparable to a bag of regular urea. While conventional urea efficiently
provided 30-40 per cent nitrogen to plants, the remaining is lost owing to
vaporization and chemical changes between water and soil movement,
erosion, and other factors. In contrast, the effectiveness of the nano urea liquid
is over 80 per cent (IFFCO, 2022).
According to Ramesh Raliya, the creator of nano urea and current
consultant for Indian Farmers Fertilizer Cooperative Limited (IFFCO), their
technique employs "organic polymers" to maintain the "nano" particles of
nitrogen stable and in a state that may be sprayed onto plants. The nitrogen in
nano urea can efficiently fulfil the needs of crops, increasing agricultural
yields by an extra 8 per cent. Nano urea is also incredibly environmentally
friendly. Traditional urea pollutes the atmosphere and water supplies, harming
biodiversity, soil, and human health. While nano urea helps to safeguard the
environment by promoting efficient usage, it also improves crop health. Nano
urea will help to lower India's import costs. Total consumption and production
are 191.01 and 136.85 lakh tonnes, respectively which was not fulfilling the
farmer’s demand (Pocket book of agricultural statistics, 2020). Thus, the
balance was met by imports. If farmers use nano urea, it will aid in lowering
reliance on imports. In this way, it may also accomplish the slogans of self-
reliant agriculture and "Self-reliant India".
Nanotechnology is being employed in many areas of development, so the
agricultural industry is not immune from it. IFFCO agricultural experts used
the same technology to develop a product that currently contains a bag of urea
in liquid form in a container of 500 ml. Crop production is increased owing to
the use of nanomaterials, which also effectively control nutrient supply to

Page | 7
plants and specific locations, ensuring that agrochemical consumption is kept
to a minimum.
The surface area to volume ratio of a nano urea liquid particle, which
ranges in size from 20 to 50 nanometers, is 10,000 times higher than that of
regular urea (IFFCO, 2022). Because of its ultra-small size and surface
characteristics, nano urea liquid is more efficiently absorbed by plants when
sprayed on their leaves. One urea particle weighs the same as 55000 nano urea
particles. The nano urea size is ‘1 nano meter’, which is ‘one billionth’ of ‘one
meter’. And 1 inch has 25.4 million nanometers.
Urea made from nanotechnology to enhance the potency of the nutrients
for the crops is termed as nano organic compound. The nano organic
compound liquid can replace standard organic compounds and will curtail
their demand by a minimum of 50 per cent. The Nano organic compound
liquid is effective and economical for plant nutrition, which will increase
production with improved organic process quality. According to IFFCO, the
nano organic compound liquid will also positively impact the underground
water standard, vital in reducing warming with an impression on temperature
change. It'll also facilitate and make the crops stronger, healthier and defend
them from the lodging impact.
Certification of nano urea
IFFCO Nano Urea adheres to OECD (Organisation for Economic Co-
operation and Development) testing guidelines (TGs) as well as the
Department of Biotechnology, Government of India's recommendations for
testing of nano Agri-inputs (NAIPs) and food products. Nano urea has been
independently evaluated and approved by National Accreditation Board for
Testing and Calibration Laboratories (NABL) and Good Laboratory Practice
(GLP) recognised laboratories for bio-efficacy, biosafety-toxicity, and
environmental compatibility. IFFCO nano fertilizers meet all the national and
international guidelines related to nanotechnology or nano-scale Agri-inputs.
With the inclusion of nano-fertilizers such as Nano Urea in Schedule VII of
the FCO1985, IFFCO has initiated manufacturing so that farmers might
eventually profit from the boon of nanotechnology. It will be a step toward
self-reliance in terms of ‘ATMANIRBHAR BHARAT’ and
‘ATMANIRBHAR KRISHI’ because of nano fertilizers (IFFCO, 2022).
Features of nano urea
IFFCO Nano Urea (Liquid) is a one-of-a-kind fertilizer created by IFFCO
for the first time in the world and certified by the Government of India.
Nitrogen is successfully delivered to the crop by spraying nano urea on the

Page | 8
leaves during necessary periods of development, increasing yield while
protecting the environment.
• Because of its ultra-fine structure and superficial properties, IFFCO
Nano Urea is easily absorbed by plants when sprayed on leaves.
These particles reach the areas of the plant that require nitrogen and
offer a balanced amount of nutrients.
• Nano urea is more affordable than solid urea. The price of a bottle of
nano urea (500 ml) has been set at Rs 240, which is 10 per cent
cheaper than a bag of conventional urea.
• This liquid urea has been found to be effective and efficient for plant
nutrition, increasing production while improving nutritional quality.
The efficacy of one bottle of Nano Urea (500 ml) is equivalent to one
bag of urea.
• It is a viable choice for farmers interested in climate change
mitigation and smart agriculture.
• It lowers gaseous emissions that contribute to environmental
pollution and climate change and the loss of nutrients from
agricultural fields through leaching, lessening the environmental
impact.
• More output is generated with less amount of input. Reduction in the
urea or other nitrogenous fertilizers without affecting the yield. It
reduces the requirement for urea by at least 50 per cent.
• Its transportation and storage are cheap and easy.
• Crop quality is enhanced by it.
• Nano urea contributes to the improvement of groundwater quality.
• The usage of nano urea can result in an extra 8 per cent boost in crop
output.
• It improves the efficiency of fertilizer use.
• Simple to use; mix with water and spray on plant leaves.
• Boost the fruit and vegetable quality.
• It is safe for the land, water, and environment.
• Safe for plants, wildlife, and humans.
• Because of their tiny size, plants can absorb and utilise more
efficiently.

Page | 9
 • Controlled utilisation and even distribution in plant cells.
• Increase the revenue of farmers. Less expensive than conventional
urea Farmers' income rises when input costs are decreased.
• In the urea sector, this will enable India to become independent.
• It plays an essential role in reducing the effects of global warming.
By enhancing the quality of the land, air, and water, environmentally
friendly products can help combat global warming challenges and
achieve the UN SDGs.
Chemical composition
Packed urea has 46 per cent nitrogen chemically; therefore, a 45 kg bag
contains around 20 kg of nitrogen. Nano Urea, in contrast, has a nitrogen
content of 4 per cent in 500 ml bottles (or approximately 20 gm).
How to use
• Nano urea solution of 2-4 ml per litre of water should be sprayed on
the standing crop.
• Nano urea can be used at the rate of 2 ml per litre of water in crops
with a low requirement of nitrogen and up to 4 ml in crops with a
high requirement of nitrogen.
• Nano urea can be used twice in cereals, oil, vegetable, cotton, etc.,
and once in pulse crops.
• The first spraying can be done after 30-35 days of germination or 20-
25 days after transplanting and the second can be done 20-25 days
after 1st spray or one week before flowering or budding.
• For one acre of field, the amount of water per sprinkle is about 125
litres at a time.
Usage guidelines and precautions
• Shake the bottle well before use.
• Use a flat fan or cut nozzle for spraying nano urea.
• Spray in the morning or evening when there is no strong sunlight,
wind, or dew.
• If it rains within 12 hours of spraying Nano Urea, it is
advisable to spray again.
• If required, Nano Urea can be easily used with Bio-catalysts

Page | 10
 (Sagarika), 100 per cent water soluble fertilizers and most
agricultural chemicals, but always do a jar test before
mixing and spraying.
• Nano Urea should be used within 2 years from its manufacture date
for better results.
• Although Nano Urea is toxin-free, it is advised to use a face mask
and gloves while spraying the crop for safety reasons.
• Please keep it in a cool, dry location away from children and pets.
Mechanism of uptake
Nano fertilizers have a greater surface area and particle size that is smaller
than the pore size of roots and leaves. This can promote penetration into the
plant from the applied surface and improve the efficiency of the nano
fertilizer's absorption and nutrient usage. Reduced particle size results in a
higher specific surface area and a number of particles per unit area of a
fertilizer, which provides more ability to interact with nano-fertilizers,
resulting in greater nutrient penetration and absorption (Liscano et al., 2000).
These nano-scale properties aid uptake in plants (Dhoke et al., 2013). The cell
wall's pore diameter ranges from 5 to 50 nm (Fleischer et al., 1999). Hence,
only nanoparticles or nanoparticle aggregates with a diameter smaller than the
cell wall pore diameter might readily pass through and reach the plasma
membrane (Moore, 2006). The size of a nano urea particle is about 20 to 50
nanometers and has approximately 10,000 times more surface area to volume
size than granular urea. They are moved to different plant sections after
entering the plant via phloem cells and plasmodesmata (40 nm diameter).
They can bind to carrier proteins through aquaporin, ion channels and
endocytosis (IFFCO, 2022). Furthermore, “due to ultra-small size and higher
surface properties of nano urea, it gets easily absorbed by the plants when
sprayed on their leaves”.
The nanoparticles having a size below 5 nm follow the cuticular pathway
and nanoparticles having greater size follow the stomatal track (Eichert and
Goldbach, 2008) and are finally transported to the conducting system, which
helps in quick and easy absorption of nutrients by leaves as they 100 per cent
soluble (Fernández and Eichert, 2009). Nanoparticles enter into the
intercellular spaces of the cell through an apoplastic pathway. In contrast, Rico
et al. (2011) hypothesized that the symplastic (through cytoplasm) route is a
more organized and regulated pathway for the movement of engineered
nanoparticles into plants.

Page | 11
 The phloem system transported these nanoparticles once they entered the
stomata. The phloem comprises of live vascular tissues that transport
photosynthetic products such as sucrose, protein, and certain mineral ions for
plant development. After that, they reach the location of nitrogen demand in
the plant and are released in a regulated manner. The site-specific and
regulated release of the active component in nano form decreases excess run-
off and avoids eutrophication and residual pollution. Fertilizers embedded in
nanoparticles will improve agricultural plant nutrient availability and
absorption (Tarafdar et al., 2012c).
Achievements of using nano urea
A field experiment is conducted by Kannoj et al., 2022a to study the effect
of nano urea vs conventional urea on nutrient content, uptake and economics
of black wheat (Triticum aestivum L.) along with biofertilizers at Udaipur
(Rajasthan). They have found that the maximum N, P, K, Fe and Zn content
and uptake in grain and straw were recorded under treatment N3 (50 per cent
conventional urea + 50 per cent of nano urea fertilizer) as compared to the
remaining treatments. The utmost net return (Rs. 109637 ha-1) and benefit-cost
ratio (2.37) were also obtained under this treatment. So nano fertilizer
application proved more practical and efficient in improving nutrient content,
uptake and economics of black wheat compared to conventional fertilizer.
Similarly, Patidar et al., 2022 found that the utmost nitrogen content in grain
(1.907%) and straw (0.490%) and the highest economic returns were fetched
by under the application of RDN + two sprays of nano urea at tillering and
jointing. An experiment performed by Kannoj et al., 2022b also revealed that
appreciable enhancement in the protein and chlorophyll content was recorded
under treatment N3 (50 per cent conventional urea + 50 per cent of nano urea
fertilizer).
Velmurugan et al., 2021 found that nano urea spray and the basal
application of RDF resulted in 15-21 per cent higher rice yield. Along with
this, the increased efficiency of nano urea resulted in saving nitrogen
fertilizers to 25-34 per cent. Samui et al., 2022 revealed that the adoption of
100 per cent RDN + foliar spray of nano-urea @ 4ml/L, twice at the knee and
tasselling stages of the maize crop, was superior in influencing morphology,
yield attributes and yield of rabi maize. It was found that the application of
20kg N ha-1 + 2 sprays of nano urea @ 4 ml L-1 could be an economical and
sustainable strategy to optimize the use of nitrogen in finger millet in southern
Odisha and appreciably improves the growth, yield parameters and yield of
finger millet (Samanta et al., 2022).

Page | 12
Some concerns about nano urea
• The lowest concentration is already present in urea, which is also
water-soluble when absorbed.
• The precise mechanism by which even smaller nano urea particles
can increase nitrogen absorption efficiency is unknown.
• Scientists are also uncertain about the product's potential to lessen
farmers' dependency on urea.
• Because a basal dose of nitrogen from urea is necessary throughout
the early phases of crop growth, traditional urea cannot be avoided.
Limitation of nanofertilizer
1) Nanomaterial phytotoxicity is an issue since different plants respond
differently to various nanomaterials in a dose-dependent manner.
2) Nanomaterials are very reactive because of their minute size with the
enhanced surface area, so this raises safety concerns for farmworkers
because they may be xenobiotic to the body.
3) Some studies have reported the phytotoxic effect of nanoparticles in
plants, depending on species, dose and application method and their
type.
Conclusion
Science and technology have taken many leaps forward with the
introduction of nanotechnology. Numerous issues facing farmers and the
environment will be resolved by its application at the field level. As well as
improving agriculture's sustainability, it will aid in preserving the environment
and natural resources. IFFCO’s Nano Urea is a massive step towards
sustainable agriculture and food systems, including precision and innovative
farming. It has the potential to revolutionise the whole agricultural sector
throughout the planet. The nano fertilizer would be more environmentally
friendly than traditional urea while lowering input, logistical, and storage
expenses. It is intended to phase out urea granules, which are now one of the
most extensively utilised fertilizers in farmlands worldwide. However, in
several other circumstances, it has not been shown to compete with traditional
fertilizers to boost crop growth and yield metrics; thus, more extensive
investigations and research are needed to analyze the influence of nano urea.
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Page | 13
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The effect of nano urea vs conventional urea on the quality and
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R. 2022a. Effect of nano urea vs conventional urea on the nutrient content,
uptake and economics of Black Wheat (Triticum aestivum L.) along with
biofertilizers. Biological Forum-An International Journal, 14(2a):499-
504.
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Amami, R. 2022. Comparative effects of conventional and nano-enabled
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Page | 18
 35.

Chapter - 2
Disease Resistance in Insects

Authors
Jasra Bano
Ph.D. Scholar, Division of Entomology, Sher-e-Kashmir
University of Agricultural Sciences and Technology, Kashmir,
India
Qadrul Nissa
Ph.D. Scholar, Division of Entomology, Sher-e-Kashmir
University of Agricultural Sciences and Technology, Kashmir,
India
Jamal Ahmad
Professor, Division of Entomology, Sher-e-Kashmir University
of Agricultural Sciences and Technology, Kashmir, India

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Page | 20
 Chapter - 2
Disease Resistance in Insects
Jasra Bano, Qadrul Nissa and Jamal Ahmad

Introduction
Insects are the earliest and most diverse taxon of animals acounting for
more species than all other animals put together because of their high
reproductive potential and varied niche. Though cellular reactions like
phagocytosis, nodulation and encapsulation and humoral reaction through
secretions of antibacterial proteins are known for bacterial and fungal
invaders, little is known about insect defense against insect virus infection.
Insects are r-selective because of their short lifespan and high rate of
reproduction. Further insects are the most successful group of animals that
exist in a myriad of environment where the potential for infection by
microorganisms and parasites is great. Insects have also demonstrated a
considerable ability to develop resistance to conventional insecticides and
more than 645 species of insects and mites have already developed resistance
to one or more chemicals. As a part of survival stratergy, insects have evolved
numerous effective resistance and defence mechanisms to most of the
conventional chemical insecticides with possession of genes for high levels of
oxidases, esterases, glutathione-s-transferases, ‘insensitive’
acetylcholinesterases (AChE), and nerve insensitivity to pyrethroids.
Similarly, an insect species susceptible to a pathogen become resistant to these
microorganisms.
Insects that are susceptible to a pathogen can show resistance to various
entomopathogens and try to resist infection through morphological,
behavioural, developmental (like maturation immunity), physiological,
nutritional, biochemical and molecular genetic mechanisms, etc. In order to
appreciate the insect defence and its role in microbial control of insect pests
by conventional agricultural entomologists and applied insect pathologists,
some preliminary information on different aspects of immunity, viz, passive
and active defence mechanisms against foreign invaders has been presented.

Page | 21
Insect immunity
Insects donot possess the ability to produce antibodies
(immunoglobulins) and donot use immunoglobulin as recognition molecules
in the classical sense, against foreign antigens and hence antigenic memory
appears to be lacking, i.e. non-memory type. Further they donot produce
alpha/beta interferons in response to viral infections. In general, insect
community consists of cellular and humoral reactions. The defence
mechanism in insects is classified into two broad groups. The first one is non-
specific immunity, which consists of structural and passive barriers like
cuticle, gut physiochemical properties and peritrophic memberane. The
second one is a specific immune system involving cellular and humoral
immunity. Cellular reactions include phagocytosis, nodulation (haemocyte
aggregation) and encapsulation, especially with reference to bacteria, fungi
and protozoa including nematode invaders. On the other hand, humoral
reactions involve activation of prophenol oxidase cascade and induction of
immune proteins such as lysozymes, lactins and anti-bacterial and antifungal
proteins.
Non-specific immunity in insects
In insects, melanin pigments and their precursors are important as
structural and protective components of the cuticle. Melanogenesis involves
the formation of melanin pigments and toxic by-products from the action of
phenoloxidase (PO) on quinone precursors, and occurs primarily in the
cuticular structures, midgut epithelium and haemolymph where it performs
dual roles in hardening and darkening the cuticle and in immune defence. Such
defences include nonself-recognition and humoral encapsulation of invading
organisms. The presence of melanin and melanogenesis intermediates in the
insect cuticle not only limits the growth of certain fungi, possibly by acting as
a physical barrier, but also suppresses synthesis of fungal cuticle-degrading
enzymes, thus further impeding cuticle penetration.
Stages of disease development in insects
The cuticle as an infection barrier Fungal germlings are continually
moving through different environments during cuticular penetration. They
respond to these changes by invoking adaptive biochemical processes and
cellular differentiation to form a series of specific morphological structures.
For example, germ tubes of Metarhizium anisopliae develop appressoria
(located at the cuticle surface), infection pegs (in the epicuticle), penetrant
hyphae and penetrant plates (in the procuticle), and yeast-like hyphal bodies
(blastospores) for dispersal through the hemocoel. These morphological

Page | 22
transitions suggest that germlings. are constantly sensing their environment
and adjusting in order to colonize insect tissue and counteract potential host
responses. Infection structures probably evolved as a mechanism by which the
pathogen overcomes host barriers.
Cuticular barriers have yet to be completely characterized for a single
insect species, but available data suggest that most, if not all, barriers can be
part of a typical resistance response acting in sequence or simultaneously
Pathogenesis is initiated by adhesion of a conidium (spore) to the insect
cuticle. Secretion of an adhesive mucus as the conidium swells during pre-
germination development supplements the initial hydrophobic interactions
between the conidium and the cuticle surface. An infection can be aborted on
the epicuticle if a factor essential for a phase of adhesion, microbial
development, or pathogenesis is absent. Specifically, infection may be
prevented by low humidity (fungi require water for germination and extension
growth), an inability to utilize available nutrients on the cuticle surface, or
absence of factors necessary for recognition of a susceptible host or penetrable
infection site. The effects of surface topography on appressorium formation
were studied using Manduca sexta cuticles and plastic replicas of the cuticular
surface. Appressoria were only produced after extensive growth over the
microfolds of the cuticle surface of early (1 day) fifth-instar larvae. These
microfolds interfere with adhesion so that the fungus does not receive
appropriate induction signals from the surface. In contrast, germination on the
comparatively flat surface of 5-day fifth-instar larvae allows appressorium
formation close to the conidium. For some systems, the failure of fungi to
invade insect cuticle has been attributed to the presence of inhibitory
compounds (phenols, quinones, and lipids) on the cuticle surface. However,
only circumstantial evidence indicates that any of these compounds are
involved in disease resistance. Penetration of the epicuticle is either by
infection pegs produced from the underside of appressoria or by direct entry
of germ tubes.
Once the epicuticle is breached, progress by the penetration peg through
the cuticle may be more or less direct via penetrant hyphae, or penetrant
structures may extend laterally, producing penetrant plates. These lateral
expansions can cause fractures that favor penetration and may facilitate
dispersal of the pathogen's cuticle-degrading enzymes. The procuticle acts as
a physical barrier by its impermeability to pathogen secretions, its resistance
to degradation by many pathogen enzymes, and its mechanical resistance to
penetration. The degree of resistance depends on the combined effects of the
cuticle's thickness the tensile strength imparted to the cuticle by the system of

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chitin lamellae, and the degree of cuticle hardening by sc1erotization. Insects
with heavily sclerotized body segments are usually invaded via arthrodial
membranes or spiracles.
Tritrophic level interactions
Development of fungal pathogens within hosts can be influenced not only
by immune reactions of hosts, but also indirectly by host food. For example,
leafcutter bees, Megachile rotundata, reared on natural provisions were
generally less susceptible to Ascosphaera aggregata as compared with bees
reared on artificial provisions. The majority of studies of tritrophic-Ievel
interactions have compared the impact of host-plant species on development
of fungal pathogens within herbivorous insects. The few in vitro studies have
demonstrated that the addition of glycoalkaloids or a diversity of plant extracts
to media can inhibit growth of B. bassiana and N. rileyi. Adult chinch bugs,
Blissus leucopterus leucopterus, inoculated with B. bassiana, demonstrated
higher mortality when fed wheat, barley, or artificial diet compared with com
or, sorghum. In addition, few cadavers of chinch bugs that had eaten com or
sorghum produced conidia, demonstrating an additional inhibitory effect of
these foods, presumably resulting from fungistatic secondary plant chemicals.
When the glycoalkaloid a-tomatine was added to artificial diet, development
of N. rileyi in Helicoverpa zea was partially prevented at LC50 and was
inhibited at LC90. Little is known of how an insect recognizes a fungus as
nonself so that its immune responses are initiated. The suggested role of
phenoloxidase activation in recognition is ambiguous because recognition
may occur in the absence of melanization. Recruitment factors have been
described that stimulate phagocytosis, deplete plasmatocytes, mobilize servile
hemocytes, and promote nodule formation. An opsonin (a galactosebinding
lectin) induced in the hemolymph mediates cellular recognition of Nomuraea
riley; blastospores. Cell walls of N. riley; hyphal bodies, the stage successfully
living within the host hemocoel, contain few /3- 1,3 glucans, while mycelial
fragments with high levels of /3-1,3 glucans are recognized and encapsulated.
In contrast, the insect phagocytes can nonspecifically recognize N. rileyi
conidia. The charge and wettability of a fungal surface apparently regulates
hemocyte adhesion in several species. The main reaction in the cellular
antifungal defense mechanism is encapsulation of the fungus, which is rapidly
melanized. During encapsulation, granulocytes are attracted to the fungus and
may engulf it (phagocytosis), then plasmatocytes are recruited and form a
pseudo tissue in concentric layers, thus differentiating a granuloma (nodule)
in which the fungus may be lysed. Encapsulation only provides protection for
hosts against weakly virulent pathogens. In the case of hypervirulent strains

Page | 24
of a fungal species, hosts are either unable to form typical granulomas or fungi
overcome the encapsulation and continue to grow. For example, the successful
development of B. bassiana within hosts is based on simply overcoming the
host hemocyte response; numbers of granulocytes are dramatically reduced 3
days after fungal challenge. Hung et al propose that the cellular-defense
response is the initial target of metabolites produced by B. bassiana. These
metabolites block the recruitment of hemocytes required for nodule formation,
although the initial recognition and phagocytosis responses remain functional.
Genetic diversity in host-pathogen systems
Antigens raised against fungal proteins can differentiate fungal species
but frequently not strains within a species. Allozymes provided the first
unambiguous markers available in sufficient numbers to enable reliable
genetic studies of entomopathogenic fungal species.
Several enzyme systems are polymorphic and sufficiently well resolved
for reliable genetic studies of M. anisopliae, B. bassiana, and the E. aulicae
species complex. Random amplification of polymorphic DNA (RAPD) and
restriction fragment length polymorphisms (RFLPs) of genomic DNA have
provided additional markers for population analysis. Other molecular
approaches applied recently include the analysis of rRNA sequences to
distinguish between Tolypocladium and the genus Beauveria and contour-
clamped homogeneous electric field (CHEF) fractionation of M. anisopliae
chromosomes to distinguish different isolates. RFLP probes and isozymes
have been used in concert to evaluate strains of the E. aulicae species complex
causing epizootics across the northeastern United States in gypsy moth (L.
dispar) populations. This fungus had never been reported from North
American gypsy moth before, although there was an attempt to introduce it
for biological control in 1910-1911. Results from these biochemical
techniques agreed that this northeastern pathogen was identical to E.
maimaiga, a member of the E. aulicae species complex from Japan, and
differed from indigenous strains of this complex. Intraspecific variability of
fungi has often been characterized by differences in pathogenicity. However,
host insect populations are not genetically static in their responses to fungal
pathogenicity. Among aphid populations, clones resistant to certain isolates of
P. neoaphidis are distributed in specific areas of Australia, coexisting among
susceptible clones at low levels. The evolution of this variability is somewhat
puzzling because it is believed that only a small population of these aphids
was initially introduced to Australia and these aphids reproduce only
asexually. Presumably, genetic variation in host susceptibility could have
arisen through one or more mutations or extranuclear inheritance mechanisms.

Page | 25
Coexistence of clones is partially maintained by differential temperature
optima and dispersal capabilities. In North America, variation in susceptibility
to one strain of P. neoaphidis has been found among 60 clones of pea aphid,
Acyrthosiphon pisum, originating from within the same field (K Hural,
personal communication). Likewise, one of two clones of A. pisum was
resistant to a strain of Conidiobolus obscurus. Sibling groups of leafcutter bees
from the same original population differ in susceptibility to chalkbrood, A.
aggregata. Matings between resistant lines resulted in loss of resistance,
suggesting a polygenic basis for the resistance, which is easily disrupted by
out-crossing. These first documented cases of variability in susceptibility
among insect hosts to fungal pathogens demonstrate the potential occurrence
of coevolution between hosts and fungal pathogens.
Control potential
Control strategies that have been successful include permanent
introduction and establishment, augmentative releases, and environmental
manipulation or conservation. For all strategies, experience has highlighted
the unique nature of interactions among fungi and hosts in specific systems.
The majority of control programs utilizing fungi rely on augmentative
releases, frequently using inundative application technology similar to that
used for standard chemical pesticides. Whether inundative or inoculative,
these releases are generally considered augmentative because of the
ubiquitous nature of pathogens. Most fungal releases differ fundamentally
from applications of chemical pesticides. After fungi are released, pathogen
densities should increase through disease transmission as the pathogen
repeatedly cycles through the host population. Different fungal species are
released using varyip. g doses, differing fungal life stages, and differing
application strategies because of system-specific expectations based on
secondary transmission levels necessary for pest control. In one example,
Beauveria brongniartii was applied to soil-inhabiting grubs of Melolontha
melolontha using two different techniques, and the speeds at which epizootics
developed were compared. Barley kernels colonized by B. brongniartii and
drilled into soil resulted in control of M. melolontha in orchards that was
superior to control from chemical pesticides. The alternative strategy of
spraying swarming adults so that egg-laying females transported B.
brongniartii to breeding sites resulted in slower, possibly more host-density
dependent, development of epizootics. To control mosquitoes in rice fields,
two types of spores are applied simultaneously; application of both sexual and
asexual spores of L. giganteum resulted in both immediate and long-term
mosquito control under varying environmental conditions. The majority of

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control efforts have emphasized inundative augmentation, the use of so-called
mycoinsecticides. Inundative releases have resulted in some control successes,
notably among greenhouse pests, rice pests, spittlebugs, and pine caterpillars,
and currently, several entomopathogenic fungi are commercially available for
inundative release. Selection of an optimal strain is crucial to successful
control, e.g. a strain of M. anisopliae tolerant to low Tasmanian temperatures
has been used successfully to control Adoryphorus couloni, a pasture scarab
In addition, cost-efficient fungal mass production and effective formulations
are critical elements in development of mycoinsecticides. An underexplored
research area is the detection, isolation, characterization, and commercial
development of the toxins of entomopathogenic fungi for insect control. These
advances may be aided by recombinant DNA techniques; however, the fact
that toxins from Metarhizium spp. (destruxins) are the products of complex,
multigenic pathways will increase the difficulty of isolating and manipulating
specific toxin genes.
Conclusion
In past years, attempts to use fungal entomopathogens for inundative
releases, similar to use of synthetic chemical insecticides, have frequently
been unsuccessful, and we now realize that to harness epizootics, we must
understand which interactions are critical determinants of pathogenicity and
epizootic development interactions are critical determinants of pathogenicity
and epizootic development. The species of entomopathogenic fungi that have
significant impacts on host populations are many and diverse. Unfortunately,
few fungal-insect systems have been studied in great detail. Studies conducted
to date, however, provide frameworks for further investigation and generate
hypotheses to be tested in other host-pathogen systems. Obviously, with the
great diversity of fungi and the many routes by which they have achieved their
success as pathogens, not everything learned about a few model systems will
apply throughout other systems. Still, we are developing a knowledge base
from which to compare systems and discover commonalities in pathogenicity
and epizootiology.
References
1. Andersen SO. 2010 Insect cuticular sclerotization: a review. Insect
Biochem. Mol. Biol. 40, 166-178.
2. Boucias DG, McCoy CM, Joclyn OJ. 1982. Isozyme differentiation
among 17 geographical isolates of Hirsutella thompsonii. 1. Invertebr.
Pathol. 39: 329-3.

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3. Boucias DG, Pendland JC. 1991. Attachment of mycopathogens to
cuticle: the initial event of mycosis in arthropod hosts. 29a, pp. 101-28
4. Butt TM, Wraight SP, Galaini-Wraight S, Humber RA, Roberts DW,
Soper RS. 1988 Humoral
5. encapsulation of the fungus Erynia radicans (Entomophthorales) by the
potato leafhopper, Empoasca fabae (Homoptera: Cicadellidae). J.
Invertebr. Pathol. 52, 49-56.
6. Cerenius L, So¨derha¨ll K. 2004 The prophenoloxidase activating system
in invertebrates. Immunol. Rev. 198, 116-126.
7. Cotter SC, Hails RS, Cory JS, Wilson K. 2004 Density dependent
prophylaxis and condition-dependent immune function in lepidopteran
larvae: a multivariate approach. J. Anim. Ecol. 73, 283-293.
8. Hajek AB, Humber RA, Elkinton JS, May B, Walsh SRA, Silver JC.
1990. Allozyme and RFLP analyses confirm Entomophaga maimaiga
responsible for 1989 epizootics in North American
9. gypsy moth populations. Proc. Natl. Acad. Sci. USA 87:6979-82
10. Hajek AE, Humber RA, Walsh SRA, Silver IC. 1991. Sympatric
occurrence of two Entomophaga aulicae (Zygomycetes:
Entomophthorales) complex species attacking forest Lepidoptera.1.
Invertebr. Pathol. 58:373-80
11. Lockwood JA. 1992. Neoclassical biological control: a double-edged
sword. Soc. lnvertebr. Pathol. Newsl. 24:6-8
12. McCoy CWO 1990. Entomogenous fungi as microbial pesticides. In New
Directions in Biological Control: Alternatives for Suppressing
Agricultural Pests and Diseases. ed. RR Baker, PE Dunn, pp. 115-38.
New York: Liss
13. Riba G, Bouviar-Fourcade I, Caudal A. 1986. Isoenzymes polymorphism
in Merarhiziam anisopliae (Deuteromycotina: Hyphomycetes)
entomogenous fungi. Mycopathologia 96:161-69
14. Soper RS, May B, Martinell B. 1983. Entomophaga grylli enzyme
polymorphism as a technique for pathotype identification. Environ.
Entomol. 12: 720-23
15. St. Leger RJ, Allee LL, May B, Staples RC, Roberts DW. 1992. World-
wide distribution of genetic variation among isolates of Beaaveria spp.
Mycol. Res. 96: 1007-15

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16. St. Leger RJ, May B, Allee LL, Frank DC, Roberts DW. 1992. Genetic
differences
17. in allozymes and in formation of infection structures among isolates of
the entomopathogenic fungus, Metarhizium anisopliae. J. Invertebr.
Pathol. 60:89-101
18. So¨derha¨ll K, Ajaxon R. 1982 Effect of quinones and melanin on
mycelial growth of Aphanomyces spp. and extracellular protease of
Aphanomyces astaci, a parasite on crayfish. J. Invertebr. Pathol. 39, 105-
109.
19. St. Leger RJ, Cooper RM, Charnely AK. 1988 The effect of melanization
of Manduca sexta cuticle on growth and infection by Metarhizium
anisopliae. J. Invertebr. Pathol. 52, 459-470.
20. Stephen WP, Fichter BL. 1 990. Chalkbrood (Ascosphaera aggregata)
resistance in the leafcutting bee (Megachile rotundata). I. Challenge of
selected lines. Apidologie 2 1:209-19
21. Stephen WP, Fichter BL. 1990. Chalkbrood (Ascosphaera aggregata)
resistance in the leafcutting bee (Megachile rotundata). II. Random
matings of resistant lines to wild type. Apidologie 21:221-31
22. St. Leger RJ. 1 99 1. Integument as a bamer to microbial infections. In the
Physiology of Insect Epidermis, ed. A Retnakaran, K Binnington, pp.
286-308. Australia: CSIRO
23. Walsh SRA, Tyrrell D, Humber RA, Silver IC. 1990. DNA restriction
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24. length polymorphisms in the rDNA repeat unit of Entomophaga. Exp.
Mycol. 14:38 1-92

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Page | 30
 Chapter - 3
Nuclear Polyhedrosis Virus (NPV): Role and
Importance for Crop Pest Management

Authors
Selvam K
Assistant Professor, SRS Institute of Agriculture and
Technology, TNAU-Vedasanthur, Dindigul, Tamil Nadu, India
Shiva N
Assistant Professor, Vanavarayar Institute of Agriculture,
Makkadavu, TNAU-Pollachi, Tamil Nadu, India
Manikandan P
Senior Research, Fellow, Division of Eco-technology, M.S.
Swaminathan Research Foundation, Kannivadi, Dindigul,
Tamil Nadu, India

Page | 31
Page | 32
 Chapter - 3
Nuclear Polyhedrosis Virus (NPV): Role and Importance
for Crop Pest Management
Selvam K, Shiva N and Manikandan P

Abstract
The treatment on pests relies heavily on the Stomach Toxicosis as the
mode of action is Nuclear Polyhedrosis Virus (NPV). For the administration
of lepidopterans at the field level, it was recommended. Virion bearing a
polyhedral structure that causes tree top disease, commonly known as
wipfelkrankheit, in larva. In comparison with chemical pesticides, residue,
resistance, and resurgence were not as commonly reported in this biocontrol
agent.
Keywords: Virus, nuclear polyhedrosis virus, mode of action of NPV,
field application of NPV
Introduction
Nuclear polyhedrosis virus (NPVs) together with granulosis viruses and
non-occludedbaculoviruses, belong to the Baculoviridae, one of the 10 virus
families that context in insects. Unlikemembers of the other virus families,
baculovirus are restricted to insect species and crustaceans. Amongthe species
infected by NPVs, many important lepidopteran pests cultivated crops are
found. NPVs are characterized by rod-shaped nucleopcapsids (250-400 x 40-
70nm) containing double- strandedcircular DNA. Nucleocapsids are enclosed
within an envelope and then called virions. Is some NPVs, onlyone
nucleocapsid is present per envelope and they are therefore designated as 5
(singly enveloped) NPVs. In others, referred to as M (multiply enveloped)
NPV, 28 nucleocapsids are enclosed in anenvelope. The virions are occluded
in large (1-10 micros) cuboidal or polyhedral proteinaceous bodies, referred
to as polyhdera, polyhedral inclusion bodies (PIBs) or polyhedral occlusion
bodies (POBS). Thereplication and assembly of these bodies occurs almost
exclusively in the nucleous of infected cell.

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 Entomopathogenicor microbial organisms derived by the process of mass
production by using the living host. Resistance, residue and resurgence were
comparatively very low in the field application. In this chapter we are going
to discuss about the NPV (Nuclear Polyhedrosis Virus) which was the well
known chemical free pest control agent for lepidopterans.
Virus
The etymology of the term virus is from Latin meaning slimy liquid,
poison or stench. The early definition of the virus was based on its
submicroscopic size and obligate pathogenicity.
a) Possession of its own genetic material, which inside the host cell
behaved as part of the cell.
b) Presence of a submicroscopic infective stage, the virion that serves
as the vehicle for transmitting viral genome into a cell. “A virus is a
set of one or more nucleic acid template molecules, normally encased
in a protective coat or coats of protein or lipoprotein, that is able to
organize its own replication only within suitable host cells”.
General features of insect viruses
A feature of many insect viruses, which does not occur in viruses infecting
plants or vertebrates, is that they are occluded, the virions are embedded
within a proteinaceous body. Occlusion bodies (OBs) vary in size from about
0.5 to over 20 Am across but are all-visible under the light microscope.
Listings of viruses infecting insects are given in Virology, Directory &
Dictionary of Animal Bacterial and Plant Viruses. A large number of other
vertebrate and plant viruses also occur in insects and may replicate in certain
insect tissues. In general, the insect is considered to be simply a vector for
transmission. Viruses in the three families, Baculoviridae, Entomopoxviridae,
and Reoviridae are unique because of the presence of occlusion bodies in
which virions, at a certain stage in their development, are occluded at random.

Page | 34
(Image from [Link]
Fig: Structure of Ebola virus-Virion
The occlusion bodies contribute to the stability and persistence of the
viruses in the environment, the viral occlusions - nuclear polyhedrosis virus
(NPV) and granulosis virus (GV) - and the spheroid are shown at the scales
indicated. The spheroid shown is of comparable dimensions to the entomopox
virus reported from Choristoneuraconflicta. Viral occlusions are called
polyhedra (singular, polyhedron) for the nuclear and cytoplasmic
polyhedrosis viruses, capsule or granule for the granulosis virus, and spheroid
for the entomopoxvirus. The occurrence of viral occlusions in three distinct
families suggests that these bodies evolved independently and indicates their
powerful selective advantage in the environment. The matrix protein in the
polyhedron is known as polyhedrin, that in the capsule as granulinand in the
spheroid as spheroidin. The polyhedrin of the NPV and the granulin of the
granulosis virus (GV) belong to one group of related proteins. Virions, bar =
100 nm (shown by the Ascoviridae). Viral occlusions (polyhedron, capsule,
spheroid) may vary greatly in size.

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Virus and environment
Polyhedra are subject to environmental factors after release from
deceased larvae. They are resistant to drought, humidity, pressure and acids,
but are rapidly inactivated by sunlight. In particular ultra-violet radiation
between 280 and 320 nm is detrimental for the virus. Generally, most virus
present in foliage is inactivated within a few days after application. Small
numbers of polyhedral situated at protected sites may persist longer. In soil
polyhedral were found to retain their biological activity for periods of more
than 10 years via splashing rain polyhedra can again contaminate foliage, but

Page | 36
it has also been shown that larvae crawling through the soil pick up virus
particles which later may contaminate foliage.
Viruses have been used to control insect pests in the classical way of
biological control by inoculative release of a virus into the population or by
inundative application as a biological insecticide. In the latter case virus is
applied whenever larval populations exceed damage thresholds. The examples
of classical method are the control of Rhinoceros beetle, Oryctes rhinoceros,
a pest of palm trees in South Pacific islands by releasing virus infected adults.
Insect viral classification
Insect viruses was proposed by Holmes (1948), who listed two genera of
insect viruses, Borrelina and Morator, in the family Borrelinaceae, suborder
Zoophagineae, order Virales. The genus Borrelina contained viruses that cause
polyhedrosis, wilt, and other diseases in [Link] genus Morator had
only one virus, the cause of sac brood in the honeybee. Steinhaus (1949)
revised Holmes classification and based on the presence, absence, or types of
occlusion bodies produced by viruses. His classification had four genera:
Borrelina with polyhedral bodies; Bergoldia with granular inclusions;
Paillotella with refringent, polymorphic bodies of very irregular shapes and
sizes; and Morator with no visible pathological inclusions.
Naming and classification
The International Committee on Nomenclature of Viruses. Presently
named the International Committee on Taxonomy of Viruses) was organised
at the Eighteenth International Congress of Microbiology held in Moscow in
1966. In the initial proposal of the classification, each virus was also defined
by a cryptogram, which was subsequently eliminated because of its
inflexibility to accommodate increased information on viruses and their
properties.
Occluded viruses
Occluded viruses belong to three virus families. Within each family the
OB proteins from different viruses are related but there appears to be no
homology between proteins from different families. The OB allows these
viruses to persist for long periods in the environment and it is presumably no
coincidence that nearly all of these viruses replicate only in the larval stages
of insects that undergo complete metamorphosis. Since in many species larvae
occur for only a limited period each year, the virus must be able to complete
its replication cycle within the larval life span and then survive for many
months in the absence of susceptible host insects. In some instances, occluded

Page | 37
virus is known to have survived for decades in the absence of any host insects
and still be infectious.
Within such cells, virus replication is dependent on the host’s protein-
synthesizingmachinery, continually giving rise to variants through various
kinds of change in the viral nucleic acid”. During the early history of their
identification and use in pest management, their pathology was based only on
symptoms, giving rise to various descriptive names of disease etiology. For
example, the grasserie of silkworm was a good French descriptor of Nuclear
Polyhedrosis Virus (NPV) (Baculoviridae) infection which resulted in
liquefaction and disintegration of the affected insects. The NPV of nun moth
(Lymantria monacha) causes changes in infected larvae that gives rise to
aberrant behaviour involving larvae climbing upwards to die in the topmost
branches of trees. This was described in German as wipfelkrankheit or tree top
disease. The wilt disease of gypsy moth, Lymantria dispar, was also described
during early studies on the ecology of this insect imported into the USA.
Recognition of occluded viruses -Steinhaus (1956), that the study of the
ecology of viruses and their potential use for pest management began. The
scope of this chapter is to give a detailed classification of insect viruses with
special emphasis on the family Baculoviridae.
Baculoviruses (BVs, family Baculoviridae)
The baculoviruses have been studied intensively, initially reflecting their
potential as pest control agents and, more recently, their prominent roles as
expression vectors for a wide range of biologically active genes. The basic
characteristic of the family is the presence of a double-stranded DNA
(dsDNA) genome that is covalently closed. Bacilliform virions are composed
of nucleo-capsids that may be singly or multiply enveloped.
Occlusion body
OBs range in size from 0.3 Am to 0.5 Am in diameter in the NPVs and
0.3 Am to 0.5 Am in the GVs. Virion dimensions are in the size range (40-140
nm) x (250- 400 nm) for NPVs and (30-60 nm) x (260 x 360 nm) for [Link]
most typical symptoms are noted in the larval stages, where either whitening
or yellowing of the gut and/or the remainder of the body organs is associated
with infection and replication.

Page | 38
Image source: [Link]
Californica-multiple-and-Fang/8aee3e79e598a044a5443db20c81f36a44c4b171
NPVS
Properties of NPVs: Nuclear polyhedrosis virus (NPVs) together with
granulosis viruses and non-occluded baculoviruses, belong to the
Baculoviridae, one of the 10 virus families that coin insects. Unlike members
of the other virus families, baculovirus are restricted to insect species and
crustaceans. Among the species infected by NPVs, many important
lepidopteran pests cultivated crops are found.
NPVs are characterized by rod-shaped nucleopcapsids (250-400 x 40-70
nm) containing double-stranded circular DNA. Nucleocapsids are enclosed
within an envelope and then called virions. Is some NPVs, only one
nucleocapsid is present per envelope and they are therefore designated as 5
(singly enveloped). NPVs. In others, referred to as M (multiply enveloped)
NPV, 28 nucleocapsids are enclosed in an envelope. The virions are occluded
in large (1-10 micros) cuboidal or polyhedral proteinaceous bodies, referred
to aspolyhdera, polyhedral inclusion bodies (PIBs) or polyhedral occlusion
bodies (POBS). The replication and assembly of these bodies occurs almost
exclusively in the nucleous of infected cell.
The natural route of infection is by ingestion of polyhdera or in some
cases by virions by the insect larvae, usually the larvae are susceptible. After

Page | 39
polyhdera are ingested with the food, the proteinaceous occlusion bodies
dissolve under the alkaline conditions (pH 9) in the larval gut. The virions are
liberated and infect the midgut epithelial cells. In the nucleus of infected cells,
new virions are produced which may leave the cell and infect cells of the
haemocoel and other tissues, such as the fat body. In these tissues, occlusion
of virions in polyhedral takes place and this process continues cell lysis. One
or two days after infection, it can be detected in nuclei of infected cells.
The larvae usually die several days later after most tissues have been
infected and at death up 30% of the dry weight of a virus, deceased larva may
consist of polyhdera. The larva completely disintegrate and the polyhedral are
spilled on the foliage and soil, available for infection of other larvae. Many
predators and parasites feeding on dead and diseased larvae have been shown
to transmit and distribute the virus. In birds and hemipterans for instance,
polyhedra pass through the digestive tract without losing their activity.
Mode of entry of baculovirus into host
The infection of baculoviruses in insects generally occurs by ingestion of
occluded and possibly free virions. Transovarially, or through the spiracle, or
through cannibalism, or the ovipositor of a parasitoid. The ingested virion
infects the exposed midgut cells and not the cells of the fore- and hindguts,
which are lined with cuticle. The columnar epithelial cells of the midgut are
susceptible and the goblet cells are not infected.

Image source: [Link]

Page | 40
Cytopathology and pathophysiology
The virions invade and fuse to microvilli of the columnar epithelial cells
of the midgut Invasion begins by the attachment and fusion of the viral
envelope to the membrane of a columnar cell microvillus. The phospholipids
and ionic charges of the viral envelope are involved in the attachment. Naked
nucleocapsids enter the cell through opening formed at point of fusion of viral
envelope to cell membrane. Nucleocapsid moves towards the nucleus of the
cell- healthy columnar epithelial cell of midgut.

Page | 41
Symptoms and gross pathology
Most lepidopteran larvae infected by NPVs show no external symptoms
for 2 to 5 days after viral ingestion. Gradual change occurs in colour and luster
of the integument with an increase in opaqueness, milkiness and
[Link] haemolymph turns cloudy and milky. The larvae become less
active and lose their appetite but may continue to feed uptoa few days before
death. The larvae generally die in 5 to 12 days. The larval period is Viral
symptoms of infected Spodoptera littoralis showing slow motion larvae (A),
red color cuticle (B), hanging larvae (C) and liquefied larval body (D)
prolonged by the egt gene product of virus, which binds with ecdysone of larva
and inhibits moulting. Larvae infected immediately after hatching die in a few
days. Death in older larvae may be delayed by even 30 days. Shortly before
death, the larvae may move away from the food, disperse, or climb an elevated
location to hang from a branch or tree top by their abdominal and caudal
prolegs, as in the case of “Wipfelkrankheit” of the nun moth, Lymantria
monachal.

Image source: [Link]

Page | 42
Mass production of NPV
In case of inavailability of natural food source for mass rearing semi
synthetic diet usage is common for both Helicoverpaarmigeraand Spodoptera
lituraat laboratory level.

Image source: [Link]

[Link]
Mass rearing of Spodoptera litura can be done on castor leaves in the
laboratory.
The steps involved in the mass production of Sl-NPV are:

Page | 43
Page | 44
Mass production of Helicoverpaarmigera NPV (HaNPV)
Mass culturing of H. armigera
Nucleus culture can be also obtained from the established laboratories.
1) The culture of H. armigerais started by collecting adults in lights
traps or at endemic areas.
2) The adults are kept @ 100 pairs in oviposition cage.
3) Cage consists of cylindrical iron frame with black cloth enclosing the
frame.
4) Eggs are laid all over the inner surface of the cloth cover by the
female moths.
5) The cloth cover with eggs are removed daily.
6) These are subjected to 10% formalin for 10 minutes, and also surface
sterilized by 0.2% sodium hypochloride solution for 5-7 minutes and
treated with 10% sodium thiosulphate to neutralize the effects of
sodium hypochlorite.
7) The larva hatched from the eggs are reared in artificial diet.
8) The larva are reared in single cell multi-cavity trays to avoid
cannibalism.
9) The larva pupates in the tray itself.
10) The pupa are collected and again surface sterilized as done for eggs.
11) Then the pupa are sexed and placed over moist sponge in adult
emerging cages similar to oviposition cages.
The dose of virus is expressed as larval equivalent (LE) and one LE is 2
× 109 POB, One LE can be obtained from 3 fully grown up and virus infected
larvae.
Components used along with NPV at the time of spray in field and their
role
1) Nuclear polyhedrosis virus: They are species specific viz., HoNPV
(Helicoverpaarmigera NPV), SINPV (Spodoptera litura NPV) etc.
2) Adjuvants: Adjuvants are used to change the pH or gut
compositionchanger. The adjuvants used are charcoal (0.05%),
tannic acid (0.01%), boric acid (0.01%), Sodium carbonate (0.01%)
3) Phagostimulants: It can be jaggery or sugarcane juice, but
commonly and widely used phagostimulant is Jaggery @ 0.5%

Page | 45
 4) Surfactant: It is being used for better absorption of the compound.
The commonly used surfactant is Teepol @ 0.05%
5) UV retardant: These retardants are used to protect NPV from UV
rays damage. The commonly used UV retardant is Tinopal @ 0.01%
6) Commercially exploited viruses for insect management
Type of virus Insect species targeted
Helicoverpaarmigera NPV
NPV Spodoptera litura NPV
Autographa californica (ACNPV)
Chiloinfuscatellus GV
GV Achaea janata GV
Phthorimaeaoperculella GV
CPV Helicoverpaarmigera

Points to be remembered
ELCAR (H. zea NPV/SNPV), the first commercial viral pesticide which
was marketed by Sandoz Companyin 1970s, was made as wettable powder
spray. 4-6 days (Incubation period) lapse between the time ofinsertion of virus
and death of the host. Dose of virus is usually expressed as Larval equivalent
(LE). One LE is 6 x 109; POB (Polyhedral Occlusion Body) and this one larval
equivalent is obtained from threematured virus infected larvae. In India, till
date only Nuclear Polyhedrosis viruses is being used for insect control.
Majority of theentomopathogenic virus commercially registered products in
India are NPV based and specifically NPVof Spodoptera litura and NPV of
Helicoverpaarmigera being widely used for management of
Helicoverpaarmigera, Spodoptera litura in pulses, vegetables, cotton,
oilseeds etc.
The Indian standard specifications for baculovirus registration are
Viral unit: NPVs (Helicoverpa; Spodoptera): 1 x 109; POB/ml or g,
(min). The NPV dose commonly recommendedin the field for management of
pests is 250-500 LE/Ha
Reasons for not commercially developing virus products in large
numbers
1) Relatively high production and registration costs.
2) Small potential market.
3) Other problems are short persistence on crop because of UV-
inactivation and their relatively slowspeed of kill. (Larvae are not
likely to die or stop feeding within 3-4 days after virus application).

Page | 46
Point to be remembered while using NPV in field
1) NPV preferably has to be used in evening hours to protect from UV
light
2) NPV has to be used againstearly instars larvae (1-3) to be more
effective as compared to late instars larvae (4-5th).
3) Few commercially available NPV formulations are Helicide
/Heliocel/Biovirusas HelicoverpaNPV and SpodocideLitucide
/Biovirus S as Spodoptera NPV
4) Baculoviruses are the most common and most widely studied group
of viruses pathogenic for insects. Baculoviruses are species or genus
specific, except agrapha californica nuclear polyhedrosis virus.
5) Viruses are obligate parasites hence need host for multiplication.
These are composed primarily ofdouble-stranded DNA. An infective
baculovirus particle (virion) is protected by protein coat called
apolyhedron.
6) The genus NPV is characterized by the presence of polyhedral
shaped viral occlusions (polyhedra)containing randomly occluded
viral particles. Polyhedroses are characterized by the formation
ofpolyhedron shaped inclusion bodies in the infected tissues of the
host.

Image source: [Link]

[Link]

Page | 47
Uses
NPVs of Amsactaalbistriga and A. moorei are used on groundnut for the
control of red hairy caterpillars
7) H-NPV is used for the control of Helicoverpaarmigera on chickpea
and groundnut
8) SI-NPV is used for the control of Spodoptera litura on tobacco,
groundnut black gram and cotton
Company Product name Formulation Dosage
Biotech International Ltd., New 0.6 to 1
Biovirus H Liquid formulation
Delhi mL/L
SomPhytopharma (India) Ltd.,
SOMSTAR-Ha Liquid formulation 1 mL/L
Hyderabad
Pest Control (India) Pvt. Ltd., 100
Heli-cide Liquid formulation
Mumbai mL/acre
Pest Control (India) Pvt. Ltd., 100
Spodo-cide Liquid formulation
Mumbai mL/acre
Biotech International Ltd., New 250 to 500
BIOVIRUS-H Liquid formulation
Delhi mL/ha
Biotech International Ltd., Noida BIOVIRUS-S Liquid formulation 1 mL/L
Spodopterin,
Ajay Biotech, Pune Liquid formulation 1 mL/L
Heliokill

Recommendation
Spodoptera NPV: 1 x 109 POBs/mL; 200 litres of water; 1 kg of jaggery;
0.1% teepol (to besprayed during evening hours).
Helicoverpa NPV: 1 x 109 POBs/mL; 200 litres of water; 1 kg cotton
seed powder; 0.1% teepol (to be sprayed during evening hours)
Field application
1) Spray NPV 2-3 times, as soon as the young instars of pest are seen in
field or approximately during flowering period.
2) Spray virus using high volume sprayer (200 litres of H2O) during
non-rainy evening 2 hours.
3) Add 1 kg of crude sugar or molasses/ac as phagostimulant.
4) Add 200 mL of teepol or soap solution as sticking agent.
5) Add 250 g of cotton seed powder for Helicoverpa.
6) NPV also compatible with chlorpyriphos can be applied when too
many pests are seen in same field or overlapping of generation of the
target pests.

Page | 48
References
1. L woff. A and Tournier. P (1971). Remarks on the classification of
viruses. In Comparative Virology. Academic Press, 1-42.
2. Matthews R.E.F. (1991). Plant virology, third edition. Academic Press,
London, G.B.
3. Evans, H.E., Entwistle, P.E (1987). Viral diseases. In Epizootiology of
insect diseases (eds J. R. Fuxa & Y. Tanada) 257-322. Wiley, New York.
4. Hull H.F. Blumhagen J.D. Benjamin D. and Corey. L (1984). Herpes
simplex viral pneumonitis in childhood. The Journal of
pediatrics, 104(2):211-215.
5. Rohrmann, G.F. (1986). Polyhedrin structure. Journal of General
Virology, 67(8), 1499-1513.
6. Summer M.D. and Egawa K. (1973). Physical and chemical properties of
Trichoplusiani granulosis virus granulin. Journal of Virology, 12(5):
1092-1103.
7. Smith G.E. and Summers M.D. (1981). Application of a novel
radioimmunoassay to identify baculovirus structural proteins that share
interspecies antigenic determinants. Journal of Virology, 39(1): 125-137.
8. Bilimoria, S.L. and Arif, B.M. (1979). Subunit protein and alkaline
protease of entomopoxvirus spheroids. Virology, 96(2): 596-603.
9. Rohrmann, G.F., Pearson, M.N., Bailey, T.J., Becker, R.R. and
Beaudreau, G.S. (1981). N-terminal polyhedrin sequences and occluded
Baculovirus evolution. Journal of Molecular Evolution, 17(6): 329-333.
10. Holmes, F.O. (1948). Order Virales, the filterable viruses. The filterable
viruses. In Bergey's manual of determinative bacteriology. Williams and
Wilkins, Baltimore, 6: 1225-1228.)
11. Steinhaus, E.A. (1949). Nomenclature and classification of insect
viruses. Bacteriological Reviews, 13(4): 203-223.
12. Wildy, P. (1971). Classification and nomenclature of viruses. Karger
Publishers.
13. Benz, G.A. (1986). Introduction: Historical perspectives, in: The Biology
of Baculoviruses, Vol. I, Biological Properties and Molecular
Biology (R.R. Granados and B. A. Federici, eds.) CRC Press, Boca Raton,
FL, 1-35.

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14. Hofmann (1891). The somnolence (Flacherie) of the nun (Liparis
monacha): In addition to e. Attachment: Lecture via Insecticide Fungi
weaver.
15. Jones, H.N. (1910). Further studies on the nature of the wilt disease of the
gypsy moth larvae. Annual Report of the State Forester, Massachusetts
43: 101.
16. Steinhaus E.A. (1956). Microbial control. The emergence of an idea. A
brief history of insect pathology through the nineteenth century. Hilgardia
26: 107-160.
17. Summers, M.D. (1991). Baculovirus-directed foreign gene expression.
ACS Syrup. Sen 453, 237-251.
18. Vaughn J.L. and Faulkner P. (1963). Susceptibility of an insect tissue
culture to infection by virus preparations of the nuclear polyhedrosis of
the silkworm (Bombyx mori L.). Virology, 20(3): 484-489.
19. Volkman, L.E. (1977). Autographa californica Nuclear Polyhedrosis
Virus: comparative infectivity of the occluded, alkali-liberated, and
nonoccluded forms. Journal of Invertebrate Pathology.30:102-103.
20. Bergold, G. (1947). The isolation of the polyhedra virus and the nature of
the polyhedra. Journal of Natural Research B, 2 (3-4), 122-143.
21. Wahl, W. (1909). Beiträgezur Kenntnis des Tektiten von Kälna in Skåne.
Geological Society of Stockholm Proceedings, 31 (6):471-478.
22. Harrap, K.A. (1970). Cell infection by a nuclear polyhedrosis virus.
Virology, 42: 311-318.
23. Summers, M.D. (1969). Apparent in vivo pathway of granulosis virus
invasion and infection. Journal of Virology, 4(2): 188-190.
24. Tanada, Y. Hess R.T. and Omi E.M. (1975), Invasion of a nuclear
polyhedrosis virus in midgut of the armyworm, Pseudaletiaunipuncta,
and the enhancement of a synergistic enzyme, Journal of Invertebrate
Pathology, 26: 99-104.
25. Kawanishi, C.Y. Summers M.D. Stoltz D.B. and Arnott H.J. (1972).
Entry of an insect virus in vivo by fusion of viral envelope and microvillus
membrane, Journal of Invertebrate Pathology, 20: 104-108.
26. Yamamoto. T and Tanada Y. (1978). Biochemical properties of viral
envelopes of insect baculoviruses and their role in infectivity. Journal of
Invertebrate Pathology, 32: 202-211.

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27. Mooventhan, Murali Baskaran, Sharma, Sivalingam, Sridhar, Mohan,
Pankaj Kaushal (2018) Eco-friendly Pest Management Technologies.
ICAR-National Institute of Biotic Stress Management Baronda, Raipur.
[online]. Available from: [Link] [Accessed January 7, 2023].
28. AEN 202 Management of Harmful and Beneficial Insects (2 + 1) (2015).
Course work Manual Tamil Nadu Agriculture University: 70-74.
29. T.V. Prasad (2022). Insect pathogens. Applied Entomology, 4: 159-172.

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Page | 52
 Chapter - 4
Post-harvest Technology for Fruits

Authors
Aakash
Ph.D. Scholar, CCS HAU, Hisar, Haryana, India
Mrinali Sood
Ph.D. Scholar, PAU, Ludhiana, Punjab, India
Garima
Ph.D. Scholar, CCS HAU, Hisar, Haryana, India
Deepak Kumari
Ph.D. Scholar, CCHAU, Hisar, Haryana, India

Page | 53
Page | 54
 Chapter - 4
Post-harvest Technology for Fruits
Aakash, Mrinali Sood, Garima and Deepak Kumari

Introduction
Total fruit production in India has been estimated 107 million tons and 6-
15% of total production in the country is spoiled due to lack of postharvest
handling up to consumption. India is second largest producer of fruits after
China. But in the case of developing country like India, the post harvest losses
noticed close to 15% of the total fruits production which badly affects the
availability of fruits to the consumers (Sudheer et al., 2007). Fruits that decay
quickly and are perishable (fruits with higher water content) make it easier for
microorganisms to attack. Post harvest losses in fruits rise as a result of
improper handling, storage, preservation methods, and microbial
deterioration. Due to some powerful heat or cold-resistant microorganisms,
the microbial effect is crucial in fruit spoiling. Processed or canned goods can
also suffer damage (Sharma et al., 2013). The use of postharvest technologies
can lower the quantity and quality of fresh fruit losses while also preserving
product quality until final consumption. Focus should be placed on cultivating
agricultural products with longer shelf life in order to ensure their hygiene
(Wasala et al., 2014). Produce after harvest needs the right processing and
transportation facilities. According to the resources (educational initiatives,
training programmes, and research programmes used), several studies found
that postharvest losses remain an issue and have not significantly decreased
over the previous 20 years. A thorough analysis shows that the overall post-
harvest losses (during harvesting, handling, packaging, and transport) range
from 10 to 20% of the total production. Fruit quality and shelf life after harvest
are influenced by growing methods, cultivar types, and environmental factors.
Postharvest losses and postharvest storage time are also influenced by the soil
and climate factors as well as integrated management techniques (Bachmann
et al., 2000). According to Salami et al. (2010), between 30 and 40 percent of
harvested fruits and vegetables are wasted before their consumption.
Reduction in the quality, storage duration and shelf life can be minimized with
the help of adequate storage, transportation and environment conditions (Ilic
et al., 2009). Postharvest losses are caused by a number of environmental

Page | 55
conditions including temperature, humidity and gaseous atmosphere.
Therefore, importance of Post-harvest technology lies in the fact that it has the
capability to meet food requirement of growing population by eliminating
losses making more nutritive food items from raw commodities by proper
processing and fortification. Post-harvest technology has potential to create
rural industries.
Factor affecting post-harvest losses
In general, post-harvest loss of fruits is influenced by number of factors
which includes losses due to physical, physiological, mechanical and hygienic
conditions. Other factors such as insect and mite injury, diseases which occur
due to non-infectious pathogens and pathological rots also cause post-harvest
loss of fruits. However, among the causes pathological rots are the most
serious which is followed by mechanical injury. The perishables suffer severe
damage from pathological rots in addition to mechanical damage.
Environmental factors including temperature, relative humidity, and oxygen
balance, particularly during storage, is responsible for damage. Temperature
and humidity also make fruits more susceptible to pathological attacks.
However, the loss of fruits due to physiological and biochemical damage are
closely interrelated (Sani and Alao, 2006). Therefore, post-harvest loss of
fruits can be studied as primary and secondary causes.
Primary causes of post-harvest loss
Mechanical injury
Mechanical damage of fruits is caused by careless handling during
harvesting, packing, transportation, storage etc. Some insects and birds are
also responsible for the mechanical injury in fruits (Hayatu, 2000). In many
cases, mechanical damage to fruits and vegetables, such as bruising and
cracking, makes them more susceptible to organism attack and accelerates the
rate of water loss and gas exchange. The rupture of inner tissues and cells is
frequently caused by mechanical damage to fruits as a result of pressure thrust
during transportation, which is frequently unnoticeable. Such food degrades
more quickly as it ages naturally (Alao, 2000). The likelihood that fruits
meeting maximal mechanical harm during harvesting is extremely
conceivable if correct ways of picking and harvesting are not employed. In
fruits like apples, it has soft outer skin which is highly prone to mechanical
damage. Therefore, fruits damaged are also prone to pathological attacks like
rotting (Yahaya, 2005). Gradually the quality of fruits deteriorates and the
produce fetches lesser prices in the market. Puncturing of the containers and
defective seals also leads to mechanical injury (Alao, 2000).

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Microbial action
Fruits are also prone to damage by microbial attack. The microbial
spoilage is mainly caused by fungi, bacteria, yeast and moulds. However,
fungus and bacteria-caused diseases are responsible for a sizeable share of the
losses of fruits during the post-harvest period. Fruits and vegetables are easily
attacked by these organisms because of their succulent nature (Elais et al.,
2010). However, apart from attacking fresh fruits, these organisms also cause
damage to canned and processed products. Many times, serious post-harvest
diseases occur rapidly and may cause extensive break down of the commodity,
sometimes spoiling the entire package (Yahaya, 2005). In general, fungi-
caused soft fruit rot is extremely devastating. Fungi like Alternaria, Botrytis,
Diplodia, Monilinia, Phomopsis, Rhizopus, Penicillium, Fusarium, etc. are the
most frequent pathogens responsible for rots in fruits as well as in vegetables.
Erwinia, Psudomonas and other bacteria inflict serious harm. High relative
humidity and temperature encourage the growth of post-harvest decay
organisms. Fruits and vegetables with a pH of greater than 4.5 are mostly
attacked by bacteria, whereas more acidic tissue is typically attacked by fungi
(Opadukun, 1987).
Environmental factors
Post-harvest loss of fruits is significantly influenced by environmental
factors such as temperature, humidity, composition and proportion of gases
under controlled atmospheric storage. The high temperature and relative
humidity favour the growth of micro-organisms which result in serious
damage to the produce. High temperature also increases the rate of respiration
of fruits which subsequently result in the breakdown of the inner tissues.
Additionally, high temperatures and high relative humidity speed up the decay
of fruits whereas low temperatures, especially below 5 °C, slow down the rate
of microbial attack on different crops. Tropical and subtropical fruits are most
commonly observed to suffer from chilling injury, which is caused by low but
not freezing temperatures. However, while the fruits and vegetables are kept
at the chilling temperature, the signs of chilling injury may not be noticeable;
instead, they may not be apparent until the fruits and vegetables are brought
to room temperature (37 °C). In the post-harvest environment, the role play by
relative humidity is important as temperature. The effects of temperature and
relative humidity are mainly comparable and interrelated because the capacity
of air to hold moisture varies with the temperature. The aeration in the storage
containers or in stores has its bearing on RH and hence indirectly on disease
development. Relative humidity near saturation reduces decay losses only if
the temperature is close to zero degrees Celsius. The effects of temperature

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and high RH on decay are closely connected. However, fruit and vegetable
surfaces cannot support microbial growth when the relative humidity is below
90% (Danladi, 2000). Therefore, all fruits have their own specific heat
requirement during processing. Tissue breakdown results in physical damage
from excessive or inadequate heat supply during processing, unsuitable cold
storage temperature and undesirable gaseous composition of controlled
environment during storage (Cho, 2008).
Heat treatment
Heat treatment includes blanching, hot water dip, saturated water vapor,
hot dry air and hot water rinse with brushing (Lurie & Pedreschi, 2014). Heat
treatments, also known as heat conditioning methods, can last from a few
seconds to up to four days and range in temperature from 37 to 55° C or less
than a minute in hot water up to 63 oC (Lurie & Pedreschi, 2014; Mahajan et
al., 2014). Blanching is the pretreatment method before FV are subjected to
further processing and storage (Xin et al., 2015). Blanching is used to sterilize
food by eradicating germs, inactivate degrading enzymes, maintain freshness
and colour, stabilize texture and nutrients (Xin et al., 2015). Depending on
their size, shape, and quantity, fruits and vegetables are often blanched by
submerging them for one to three minutes in boiling or almost boiling water
(James & Matemu, 2016; Vaclavik & Christian, 2014).
Secondary causes of post-harvest loss
Inadequate methods in harvesting, transportation, storage, marketing
facilities and legislation result in favourable conditions for causing of
secondary losses. Inadequate harvesting equipments and rough handling
during harvesting results in bruising and increase possibilities of contact of the
produce with the soil which may leads to contamination with micro-organisms
(Yahaya, 2005). Also extended time taken for harvesting and grading in field
exposes the fruits with field heat for longer period of time which, may
subsequently causes faster senescence and reduce shelf life. In addition, the
use of improper machinery and equipment’s in mechanical harvesting cause
serious losses. For example, mechanical harvesting of tomato has been
reported to result in more cracking (33%) than hand picking (10%). While
harvesting of fruits during rain or immediately after rains creates conditions
favourable for decay organisms. But harvesting during hotter part of the day
results in faster senescence, shriveling and wilting of fruits as compared to
produce harvested in the early morning or later afternoon (Yahaya, 2005).
Furthermore, inadequate storage facilities at the producing or marketing
centers, exposes the produce to the natural causes of losses i.e. damage by

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micro-organisms, respiration, transpiration and other biochemical reactions.
While proper storage, however, creates conditions unfavourable to these
factors. Therefore, transportation and distribution of fruits are the important
stage of post-harvest loss. Longer shipment and distribution period eventually
caused heavy losses (Alao, 2000). The earlier the harvested produce is
consumed, the minimum is the loss, as the period of senescence and organism
invasion, multiplication and damage is shortened. Only a carefully planned
and implemented marketing approach can accomplish this. Therefore, the
eventual retention or rejection of a vegetable or fruit for human eating might
be influenced by the existence or absence of appropriate regulatory criteria.
However, legal standards differ from country to country and are impacted by
the population's economic situation as well as public pressure to increase fruit
and vegetable consumption. Therefore, rigorous adherence to the law will
result in higher consumer quality and greater prices for farmers.
Control of post-harvest loss
The magnitude of post-harvest loss in fruits and vegetables can be
reduced by proper cultural operations, harvesting, transportation, storage and
pre and post- harvest treatments. These are briefly discussed here.
Cultural operations
In order to ensure normal development and increase shelf life of fruits a
good cultural operation is desirable. In perishable, preparation of the soil to a
fine tilth of porous nature is necessary to avoid root forking especially for
vegetables like root crops such as carrot and radish. Maintenance of regular
irrigation during the stage of development of fruits and vegetables are of
significant importance (Cho, 2008). When nitrogenous fertilizers are heavily
applied, fruits and vegetables typically experience faster tissue degeneration;
nevertheless, a regular supply of potassium fertilizers increases the keeping
quality of fruits and vegetables (Danaldi, 2000). Proper training and pruning
of fruit trees and removal of dead or infected branches enable the, trees to
receive proper sunlight which is essential for the colour and size development
of the fruits. While destruction cause by birds in orchards can be reduce by
using some mechanical devices in addition to good and careful supervision.
Some strategies for preventing bird damage to fruit include putting bird scarers
over trees and creating noise with a drum (Cho, 2008).
Harvesting and field handling
Consumers generally favour fresh, adequately matured, insect and
disease-free fruits and vegetables with a pleasing look, according to studies by
Hayatu (2000). In case of fruits, ripening in trees is mostly preferred. Majority

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of the fruits are harvested at proper maturity stage but quite earlier than the
ripening stage. They, therefore, ripened artificially later on as per the market
demands (Opadokun, 1987). The best time for harvesting of fruits is cooler
parts of the day and after harvest the produce should immediately be shifted
to the packing shade. However, harvesting during the hottest part of the day
typically increases the produce's field heat and speeds up wilting and
shriveling. Many harvesting aids have been designed for fruit harvesting that
prohibit the usage of ladders in favour of mobile platforms, which puts the
picker in the proper harvesting posture. However, manual fruit harvesting is
quite difficult due to the big size of fruit trees, which causes more harm to the
harvested fruits. Yet manual harvesting is easier and more efficient in the case
of high-density fruits (Alao, 2000). Equipments like buckets, sacks, baskets
and boxes are used in collection of harvested produce from the field to a
collection point, but most of these containers are unhealthy and poorly
designed, inadequately maintained and unsuitable for the harvesting and
transportation of the fresh produce. Therefore, in order to ensure good quality
of produce, the container for collection of harvested produce should be
designed in such a way that they do not cause any injury to the field operations;
also a high standard of field hygiene should be maintained. Therefore, fruits
should not be allowed to remain on the ground for any extended period of time
as it becomes a source of infection. Complete disinfection or destruction of all
disease-infected materials is required.
Cleaning
All farmers, intermediaries and consumers are extremely concerned about
the cleanliness and hygienic conditions of the produce in order to prevent both
post-harvest infections and consumer food-borne illness (Arah I.K. 2016).
Fruits and vegetables should be cleaned by using suitable methods and
materials. Sodium hypochlorite solution, thiabendazole, chlorinated water are
frequently used for cleaning or disinfection. The pH of wash solution is found
to be more effective in between 6.5 to 7.5.
Washing and grading
Fruits must be handled carefully before being packed for transportation.
Fruits treated with toxic chemicals need to be thoroughly washed before being
packaged. Washing with clean water prevent wilting and also improves the
appearance of fruits (Alao, 2000). Therefore, for good and effective marketing
strategies grading is an important component which should not be neglected.
The produce is therefore, sorted into various grades and attractive forms which
will attract good price for the produce (Hayatu, 2000). Sometimes fruits are

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graded on the basis of shape, size and colour. While fruits and vegetables such
as okra, cucumber, ridge gourd may also be graded on the basis of their
maturity, ripeness in mango and tomato fruits and general appearance.
Therefore, during sorting of fruits and vegetables, all the characters that
influence quality and appearance should be considered (Opadokun, 1987).
Packaging
Significant reduction of wastage of fresh fruits is recorded as a result good
packaging. In addition, good packaging also ensures protection against
mechanical damage, undesirable physiological changes and pathological
deterioration during storage, transportation and marketing, likewise freshness,
succulence and flavour of fruits can be maintained for a longer period through
good and effective packaging (Alao, 2000). Some inexpensive packaging
techniques and materials, such as polythene films, paper board boxes lined
with polyethylene, and other materials, may be helpful for efficiently
extending the shelf life of fruits and vegetables. Also, plastic films provide
good protection for the fruits against dry air. To prevent the physiological
breakdown of the produce, some amount of ventilation must be provided in
every package. If there is impermeability to carbon dioxide and water vapour
inside the package, there may be the development of off-flavour. While
packing for transportation, care should also be taken to avoid bruising and
damage of fruits (Hayatu, 2000). For this purpose, dry grasses or paddy straw
can be successfully used between the layers of fruits. However, packaging is
a highly innovative technology and newer techniques and material are being
added every year, both for fresh fruits and vegetables and also for processed
products. Packages can be flexible or rigid, each group having distinct
advantages and limitations. An important application of flexible film is in
wrapping the produce for low temperature (Hayatu, 2000). A mixture of gases
can be inserted in film packs that delay post-harvest losses. Recently,
thermoformed pots for aseptic filling and Tetraking R, a D-shaped cylinder
made of two webs of polystryene and expanded polystyrene material, were
launched. Sterilization is carried out by either using liquid or vapour of
Hydrogen peroxide or using ultra violet or gamma radiation.
Pre-cooling
Pre-cooling prevents premature ripening and ageing of the fresh produce.
Therefore, it is important to remove field heat of the harvested fruits,
especially when the harvesting is done during hot weather. In fruits like tomato
pre-cooling during storage can reduce physiological loss in of weight from 6
to 2.9 per cent. Therefore, effective pre-cooling can be accomplished by

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placing the produce in refrigerated trucks with forced humidified air
circulation, by placing ice in packages (Alao, 2000), by placing ice in water
and then passing through a spray of cool water by passing through vacuum
cooling (Elias et al., 2010). While hydro-cooling can be performed by
flooding the field, spraying or immersing is a quicker and more effective
control approach since water is a good medium for transferring the heat from
the produce to the cooling medium. Also, a small amount of relevant fungicide
can also be added in hydro-cooling water to for the control of fungal growth
(Danladi, 2000). Salad and other leafy vegetables are often cooled by lowering
the atmospheric pressure in hermetically sealed chamber until the reduced
vaporizing point of water produced at low pressure in the cooling chamber
cool the produce. In leafy vegetables and some fruits like grapes are usually
very difficult to cool with water or air, but they can be field packed and cooled
rapidly and uniformly by vacuum cooling (Cho, 2008).
Transportation
The transportation of perishable commodities is in the most precarious
stage (Alao, 2000). In our local markets the produce are brought either by
trolley, bus, motorcycles, or tricycle. The long distance of transportation is
mainly by rails and trucks, which is very costly. However, the main reason for
preference to road transportation against rail or truck is because it takes short
transit period. For successful marketing and maintenance of good quality
quick transport of fruits is important. The majority of losses that occur during
transportation are caused by mechanical and physical damage as well as
uncontrolled circumstances, mainly temperature and humidity (Opadokun,
1987). Therefore, to ensure reduce losses of the produce, adequate long-
distance facilities especially by rails should be developed. Also, the produce
should be properly packed and stacked in healthy and well-ventilated
containers. Control of relative humidity and refrigeration in containers is of
important. Additionally, it is advised to maintain good roads for smooth
transportation with little to no vibration and shocks. Therefore, a reliable and
efficient transportation infrastructure can significantly help to stabilize the
price variations of various commodities available in the country's numerous
marketplaces as well as to reduce the post-harvest loss of horticultural
produce.
Reducing moisture loss
One of the main advantages of consumer packaging, especially when
moisture-retentive films are employed, is the avoidance of weight loss of fresh
produce. Produce that is packaged has a longer shelf life than produce that is

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not, especially when it is sold from refrigerator display cases. According to
certain reports, moisture-proof cellulose film cut tomato weight loss in half
compared to unwrapped tomatoes (Yahaya, 2005).
Marketing of fresh produce
To prevent the loss of fruits and vegetables, as well as to be able to sell
them at a time when the market is most in need of them and to earn a fair
return on the time and money invested, a proper and effective marketing
strategy is required. However, marketing of fresh perishables presents more
problems compared to other durable agricultural products (Yahaya et al.,
2015). The interests of producers and consumers are generally very poorly
served in developing nations, the famers get less return for their effort and
money spend while the consumer pays more than what is necessary. However,
during the peak of the season when the market is in glut with a particular fruit
or vegetable a lot of wastage or loss is experienced. Also, prices are
considerably very low, and farmers get discourage and dejected. In order to
avoid overindulgence and keep the loss to a minimum, efforts should be done
(Alao, 2000; Yahaya et al., 2015).
Storage at low temperature
The main objective of storage is to extend the shelf life of fruits and
vegetables and increase their period of availability. It should, therefore, be
noted that fresh produce are living entities and even after harvest they carry
out all the vital physiological activities. Therefore, controlling respiration,
transpiration, ripening, undesirable biochemical changes, and disease
infection are the main goals of storage. Many fruits and vegetables can be
prevented to a greater extent by maintaining the post-harvest climatic
parameters of temperature, relative humidity, and air concentration of specific
gases (Danladi, 2000).
Refrigeration
The principle of refrigeration is to bring cool and take out heat from the
produce. In mechanical refrigeration, the refrigerated gas, such as ammonia,
removes heat from the chamber or storage. Therefore, the expanded gas is then
compressed to remove the heat from the compressed air over the tubes
containing the hot gases. The gas is liquefied, and the cycle is then repeated.
Fruits and vegetables can thus be safely preserved through refrigeration for a
long time (Danladi, 2000).
Cold storage
The most suitable condition for fresh fruits and vegetables in storage is
the lowest temperature, which does not cause chilling injury to the fresh

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produce (Alao, 2000). Any variation from the desired condition is detrimental.
Relative humidity of the store rooms also has a considerable bearing on the
keeping quality of the fresh produce. Therefore, control of moisture in air is
very difficult (Opadokun, 1987). Temperature and the rate of respiration are
directly correlated; the higher the temperature, the greater the rate of
respiration and the growth of organisms that cause degradation. But it should
be noted that the temperature and relative humidity requirements differ for
different fruits and vegetables. Recommended storage temperature for some
fruits: pineapple (8-10 0C), Guava (8-10 0C), Mango (7.20-8.80 oC), jackfruit
(11.70-12.30 0C). According to Williams et al., (1991) one of the most
important contribution to the storage technology is the Controlled Atmosphere
(CA) storage, were low level of Oxygen (O2) and high level of Carbon Dioxide
(CO2) and are maintained. When refrigeration is used with controlled
environment storage, the respiratory processes are slowed, which has the
potential to delay processes like softening, quality alterations and yellowing.
However, the most important aspect of CA storage is the adherence or
sensitivity of the fruits to harm brought on by an increase in CO2 and a
decrease in O2 concentration.
Ionizing radiation
The ability of ionizing radiation to eliminate pathogenic organisms
without significantly raising the temperature of the product is one of its most
important advantages. Though, it involves high initial capital involvement
(Elias et al., 2010). The Pasteurization with ionizing radiations has being
widely accepted because of its easy application it leaves no residue and require
minimal energy expenditure. The ionizing radiation prolong the shelf life of
perishables as surface pasteurization, sprout inhibition and slowing the
process of senescence. It has been found that irradiating bananas with 25-35
krads effectively delays the start of natural ripening without degrading the
fruit's quality. Additionally, the ionizing radiation performs best at 10-15
krads to stop the sprouting of potatoes, onions, and other bulbs.
Waxing
The main importance of wax coating is to reduce evaporation and
respiration (Alao, 2000). Wax-coated protective skin coating is another
technique used to extend the shelf life of fresh fruit in situations where
refrigerated storage facilities are unavailable or insufficient (Elias et al.,
2010). Fruits and vegetables such as tomatoes, okra, and mangoes have longer
shelf lives after being passed through water emulsified with mustard oil.

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Chemical treatment
Other than fungicides, a number of chemicals extend the shelf life of fruits
by delaying the ripening and senescence processes. Post-harvest treatment
with gibberellic acid markedly delayed the ripening of banana (Hayatu, 2000).
Potassium per magnate is used as an ethylene absorbent to keep the ethylene
below the threshold level. It has been found that the maleic hydrazide is
particularly effective at delaying the ripening of several fruits. At 1000-2000
ppm maleic hydrazide delays the ripening of mangoes (Yahaya, 2015).
Effect of in-package desiccants
For maintenance of the succulent and tenderness of fruits and vegetables,
partial dehydration is carried out and thereafter in-package desiccants are used
to lower the level of moisture. The shelf life of dried fruits and vegetables can
be extended by using in-package desiccants like calcium oxide or fumed silica
to reduce the moisture content of the storage environment to 1% or lower.
(Elias et al., 2010)
Importance of fungicides
Fungicides, along with a number of cultivation technique modifications
and the creation of resistant or tolerant types, are necessary for controlling the
pathogenic microorganisms that cause losses of fresh produce. Pre-harvest
spraying of fungicides can result in significant reduction of the pathogens in
the harvested produce. The best method of application of fungicides and
antibiotics is to mix them in wax emulsions (Hayatu, 2000). To stop the spread
of fungus like Botrytis, storage goods are fumigated with sulphur dioxide.
Additionally, fumigants including ethylene oxide, propylene oxide, bi-phenyl
acetaldehyde, and nitrogen trichloride can be used. It also marked inhibit the
activity of a number of micro-organisms and extend the shelf life and added
nutritive value to foods (Elias et al., 2010). However, it is recommended that
the post-harvest treatment of fruits and vegetables with poisonous chemicals
should be avoided. When required, delaying the consumption of these fruits
and vegetables for at least one or two months is recommended as they present
a health risk, especially when consumed after certain respiratory and digestive
problems (Yahaya, 2015).
Perforated films
Perforated films used for packing the fruits which kept the product safe
on the basis of the amount of escaped gases via perforated layer as well as the
gases enters from polymeric film is balanced with the throughout gases
movement rate of perforated film (Fishman et al., 1996). In the case of

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perforated film packaging, cool air has been given to keep the packaged fruits
at an appropriate temperature (Sharp et al., 1993).
Future approach
World population growing up rapidly and expected to become 10.50
billion up to 2050 means demand of fruits and vegetables ramped up. Per
capita availability of the fruits has been increased with the reduction of
postharvest losses. According to Kader and Rolle (2004), just 5% of all
research funding is allocated to reducing postharvest losses of fruits and
vegetables, whereas 95% of all funding is dedicated towards increasing
productivity. Intense research needed for minimizing the postharvest losses
because nearly half of the produced fruits have spoiled. Over the past 20 years,
far less money has been spent on post-harvest processing research and
development for horticulture products. Instead, the emphasis has been on
improving production (Kitinoja et al., 2011). Strong communication and idea
sharing between farmers, postharvest engineers, food technologists, and
market experts of fruits have been necessary for the effective elimination of
postharvest losses. This is because postharvest losses can be reduced and the
shelf life can be extended through advanced research and the use of new
scientific approach (Kader, 2005). According to a study, quality and cost of
the product have been reduced to hasten processing and increase profits. As a
result, further research focused on the processing of consumable products was
necessary to produce high-quality, cost-effective products (Ram et al., 2008).
Conclusion
Post-harvest loses of fruits occur due to lack of proper technique of
harvesting, transportation, storage and distribution. The freshness of fruits
after harvest is controlled by water content, respiratory rate, ethylene
production, endogenous plant hormones and external factors such as microbial
growth, temperature, relative humidity and atmospheric compositions.
Therefore, post-harvest loss of fruits can be considerably reduced and their
shelf life increased by careful manipulation of these factors. Implementing
crucial cultural practices, careful handling and packaging prevents
unnecessary loss. The use of appropriate chemicals at pre and post-harvest
stage may prolong the availability of fresh produce for a long period of time
by protecting them from pathogens and other environmental factors. Fruits can
also benefit from controlled environment storage and redurization at low
temperature.

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Page | 70
 Chapter - 5
Nutrient Use Efficiency

Authors
Abhishek Raj Ranjan
Ph.D. Scholar, Department of Agronomy
Anurag Singh Suman
Ph.D. Scholar, Department of Soil Science and Agricultural
Chemistry

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Page | 72
 Chapter - 5
Nutrient Use Efficiency
Abhishek Raj Ranjan and Anurag Singh Suman

Increases in population and per capita use of energy and animal products will
worsen resource depletion, pollution, and land degradation if no action is
done. This will harm the quality of our water, air, and soils and have an impact
on the climate and biodiversity. The "Nutrient Nexus," wherein decreased
nutrient losses and higher nutrient usage efficiency across all sectors
concurrently offer the framework for a greener economy to generate more
food and energy while lowering environmental pollution, calls for a new
global endeavor.
It is of great importance to produce enough food for the growing global
population (Long et al., 2015). In such a situation, the agroecosystems usually
harbour a dominant ecological force of strong fluctuations in fertilization
inputs and production outputs (Wittwer et al., 2021), in which large amount
of mineral fertilizer input is essential (Vitousek et al., 2009). However, a great
amount of literature indicates that NUE is low in most agroecosystems of the
world, generally lower than 40% and particularly in those ecosystems with
excessive mineral fertilizer input (Vitousek et al., 2009; Omara et al., 2019).
Higher or unbalanced mineral inputs may cause soil acidification and lead to
accumulation of nutrient residue in the soil profile (Guo et al., 2010; Wang et
al., 2010). Meanwhile, these issues have been proved to decrease the nutrient
cycling microbial activity and reduce nutrient utilization (Dong et al., 2014;
Xun et al., 2016). In contrast, balanced nutrient provision helps to increase the
NUE and improve plant fitness under abiotic and biotic stresses (Salim and
Raza, 2020). An extensive body of experimental and observational studies
across different types of ecosystems confirm that a combination of organic
and mineral fertilizer is the best strategy for NUE improvement, in which all
of the studies emphasize the great importance of soil and rhizosphere
microbiomes in improving NUE (Hua et al., 2020; Wang et al., 2021; Xiao et
al., 2021). Therefore, it is important to gain insights into the microbial
mechanisms behind the association between fertilization management and
NUE, hence to improve NUE and enhance crop productivity.

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 A new holistic framework of bioavailable N cycling points out that
microbial physiological traits substantially impact the entire bioavailable N
cycle in fertilized agroecosystems, which should be considered to develop
agricultural management strategies to increase N use efficiency and minimize
N pollution (Daly et al., 2021). These biological properties are responsible for
ecosystem functions and may dominate the NUE of crops (Pii et al., 2015;
Zhu et al., 2016). However, the entire community's or a selected bacterial
taxon's particular functions' microbial activity offers a valuable but
constrained knowledge of how fertilisation management influences NUE.
Understanding the underlying characteristics of microbial assemblage
changes is crucial, as are any potential downstream functional effects that can
affect NUE and soil productivity.
Scenario of nutrients
For more than a century, mineral fertilisers have supported worldwide
agriculture, resulting in increases in population and prosperity. Their
contribution to boosting agricultural yields has prevented the conversion of
millions of hectares of natural habitats to farmland. The use of nutrients in
agricultural systems that is insufficient, unbalanced, improper, or excessive is
still an issue. Low crop yields are primarily the result of nutrient mining in
developing nations, particularly in Africa. In other cases, nutrients like
phosphorus (P) and nitrogen (N) frequently leave the agricultural field because
the management techniques employed do not successfully achieve good
congruence between nutrient supply and crop nutrient demand. If left
unchecked, such losses may be significant costs to society. Hence, increasing
nutrient use efficiency continues to be a major challenge for world agriculture.
Measurement of nutrient use efficiency agronomic indices for short-term
assessment of nutrient use efficiency
Table-1 summarizes a set of simple indices that are frequently used in
agronomic research to assess the efficiency of applied fertilizer (Novoa and
Loomis, 1981; Cassman et al., 2002), mainly for assessing the short-term
crop response to a nutrient. A practical example is illustrated in Figure 1. Other
indices are sometimes used (Gourley et al., 1993; Huggins and Pan, 1993),
but they have no additional advantages for understanding fertilizer best
management practices (FBMPs). More detailed studies on the fate of nutrients
in agro-ecosystems often involve isotopes, which are particularly useful for
understanding loss, immobilization, fixation and release mechanisms.

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 Table 1: Indices of nutrient use efficiency, their calculation using the difference
method, and their interpretation.

Index Calculation Interpretation Nitrogen in cereals

• RE depends on the
congruence between plant
demand and nutrient
release from fertilizer.
RE = Apparent crop 0.30-0.50 kg/kg;
• RE is affected by the
recovery efficiency 0.50-0.80 kg/kg
application method
of applied nutrient RE = in well-managed
(amount,
(kg increase in N (U - Uo)/F systems, at low
timing, placement, N
uptake per kg N levels of N use, or
form) and factors that
applied) at low soil N supply
determine the size of the
crop nutrient sink
(genotype, climate,
• Plant density,
abiotic/biotic stresses).
• Ability of a plant to
transform nutrients
acquired from fertilizer
PE = Physiological 40-60 kg/kg; >50
into economic yield
efficiency of kg/kg in
(grain).
applied well-managed
PE = (Y- • Depends on genotype,
N (kg yield increase systems, at low
Yo)/(U- Uo) environment and
per kg increase in levels of N use,
management.
N uptake from or at low soil N
fertilizer) • Low PE suggests sub- supply
optimal growth (nutrient
deficiencies, drought
stress, heat stress, mineral
toxicities, pests).
• Ability of a plant to
transform nutrients
acquired from all sources
(soil, fertilizer) into
economic yield (grain).
IE = Internal • Depends on genotype,
30-90 kg/kg;
utilization environment and
55-65 kg/kg is
efficiency of management.
the optimal range
a nutrient IE = Y/U • A very high IE suggests
for balanced
(kg yield per kg deficiency of that
nutrition at high
nutrient nutrient. yield levels
uptake) • Low IE suggests poor
internal nutrient
conversion due to other
stresses (nutrient
deficiencies, drought
stress, heat stress, mineral
toxicities, pests).

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 • Product of nutrient
recovery from mineral or
AE = Agronomic organic 10-30 kg/kg;
efficiency of • Fertilizer (RE) and the >25 kg/kg in
AE = (Y -
applied efficiency with which the well-managed
Yo)/F or
nutrient plant uses each additional systems, at low
AE = RE x
(kg yield increase unit of levels of N use,
PE
per kg nutrient nutrient (PE). or at low soil N
applied) • AE depends on supply
management practices
that affect
• RE and PE.
• Most important for
farmers because it
PFP = Partial factor 40-80 kg/kg;
integrates the use
productivity of PFP = Y/F >60 kg/kg in
• Efficiency of both
applied nutrient or well-managed
indigenous and applied
(kg harvested PFP = systems, at low
nutrients.
product (Yo/F) + levels of N use,
• High indigenous soil
per kg nutrient AE or at low soil N
nutrient supply (Yo) and
applied) supply
high AE are equally
important for
• PFP.

F - Amount of (fertilizer) nutrient applied (kg/ha)

Y - Crop yield with applied nutrients (kg/ha)
Yo - Crop yield (kg/ha) in a control treatment with no N
U - Total plant nutrient uptake in aboveground biomass at maturity
(kg/ha) in a plot that received fertilizer
U - Total nutrient uptake in aboveground biomass at maturity (kg/ha) in
a plot that received no fertilizer
In field experiments, nutrient utilization efficiencies are either evaluated
using isotope-labeled fertilizers to measure crop and soil recovery of applied
nutrients, or by comparing crop yield and/or nutrient absorption between
treated plots and an unfertilized control (the "difference technique"; Table 1).
With increased fertilizer input (F), crop output (Y) and plant nutrient
accumulation/uptake (U) normally rise and eventually reach a ceiling.
Only situations where changes in root systems between unfertilized and
treated crops are minimal and where soil nutrient concentration is at a
generally stable state can agronomic indices accurately estimate nutrient use
efficiency. For instance, it is difficult to account for nitrogen in roots and any
net buildup of nitrogen from fertilizer in soil organic matter and its impact on

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the native soil's availability of nitrogen for following crops. This might result
in an underestimating of the applied N inputs' overall system level efficiency.
Current status of nutrient use efficiency
Nitrogen
Nearly 60% of the N fertilizers used globally in recent years 83-85 million
metric tonnes (Mt) were applied to cereal crops (Dobermann and Cassman,
2005). Globally, during the last 40 years, there has been a nearly linear growth
in grain output (slope = 31 Mt/year), cereal yields (slope = 45 kg/year), and
fertilizer N usage (slope = 2 Mt/year). regional or global levels, Because of
uncertainty regarding the real N consumption by various crops as well as crop
production figures, the only measure of N use efficiency that can be estimated
more easily is the PFPN (Table 1). However, this index cannot be calculated
with absolute precision. PFP always falls from large values at low N
application rates to smaller values at high N application rates since it is a ratio.
The types of cereal crops planted, their potential yield, the soil's condition, the
quantity and type of N applied, and the general timeliness and quality of other
crop management activities all affect the average cereal PFPN in different
parts of the world.
In many developed countries, cereal yields have continued to increase in
the past 20 years without significant increases in N fertilizer use, or even with
substantial declines in N use in some areas. This has resulted in steady
increases of PFPN in Western Europe (rainfed cereals systems), North
America (rainfed and irrigated maize), Japan and South Korea (irrigated rice)
since the mid 1980s (Dobermann and Cassman, 2005).
In developing regions, N fertilizer use was small in the early 1960s and
increased exponentially during the course of the Green Revolution. The large
increase in N use since the 1960s resulted in a steep decrease in PFPN in all
developing regions. Regional N rates on cereals range from less than 10 kg
N/ha in Africa to more than 150 kg N/ha in East Asia (Table 4) and, with the
exception of Africa, PFPN continues to decline in all developing regions at
rates of -1 to -2%/year (Dobermann and Cassman, 2005).
The shortage of information on farm-level N use efficiency in key
cropping systems has hampered efforts on designing the right N management
strategies for reducing reactive N loads and increasing farm-level profitability
(Cassman et al., 2002). On a worldwide scale, it is safe to anticipate that at
least 50% of the applied nitrogen fertilizer (N) is lost from agricultural
systems, and the majority of these losses take place in the year of fertilizer
application. However, it has also been demonstrated through research, the best

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farmers and commercial implementation of new N management technologies
that 30 to 50% increases in N use efficiency can be achieved in many crops
(Dobermann and Cassman, 2004; Giller et al., 2004).
Table 5: Average N use efficiency terms for cereals in different world regions:
literature summary of field trials conducted at research stations and averages of
selected on-farm studies.

Region/crop N rate RE15N REN PEN AEN PFPN

(kg/ha) ............................................................ (kg/kg) ...........................................
Research station trials (Stationary treatment plots) 1
Africa 139 0.37 0.63 23 14 39
Europe 100 0.61 0.68 28 21 50
America 111 0.36 0.52 28 20 50
Asia 115 0.44 0.50 47 22 54
Average 0.44 0.55 41 21 52
Maize (rainfed & irrigated) 123 0.40 0.65 37 24 72
Rice (irrigated) 115 0.44 0.46 53 22 62
Wheat (rainfed & irrigated) 112 0.45 0.57 29 18 45
On-farm studies (Non-stationary treatment plots)
Maize, USA (rainfed & irrigated)2 158 - 0.36 33 12 61
Maize, USA (irrigated)3 142 - 0.57 41 23 94
Maize, Indonesia (rainfed &
117 - 0.31 39 12 49
irrigated)4
Rice in S, E and SE Asia (irrigated)5 117 - 0.31 39 12 49
Rice in West Africa (irrigated)6 106 - 0.36 47 17 46
Wheat in North India (irrigated)7
134 - 0.34 32 11 44
RE15N - average N recovery efficiency measured with the 15N isotope dilution method.
All other N use efficiency terms - difference method, as described in Table 1.
1Research station trials summarized by Ladha et al., 2005. Most of those are
multi-year or long-term trials with stationary treatment plots.
252 sites in IL, KS, MI, MN, MO, NE and WI, 1995-1998 (Cassman et al., 2002).
332 site-years in Nebraska, 2001-2004 (Dobermann et al., 2006).
425 farms in Indonesia, 2004-2005, at N rate of 200 kg N/ha (Witt et al., 2006).
5Farmers’ fertilizer practice, 179 farms in China, India, Vietnam, Indonesia and
the Philippines, 1997-1999 (Dobermann et al., 2002).
Farmers’ fertilizer practice, 151 farms in West Africa (Wopereis et al., 1999;
6

Haefele et al., 2001).

7Farmers’ fertilizer practice, 23 farms in Uttar Pradesh, 1998-1999.

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Phosphorus
The global patterns of P supply, consumption and waste production have
become decoupled from natural P cycles (Tiessen, 1995). Global mobilization
of P has roughly tripled compared to its natural flows, and global food
production is now highly dependent on the continuing use of phosphates
(Smil, 2000). Despite the fact that most crops utilize P well, a major
contributor to eutrophication is lost P that travels to aquatic environments
downstream from agricultural regions. In developed nations, phosphorus
surpluses are common owing to the usage of fertilizers, the livestock sector,
and imports of feed and food. Contrarily, P surpluses and deficits are present
in emerging nations, as well as a substantial area of P deficient soils (mostly
in the tropics), for which the only method to boost agricultural output and
revenue is through the injection of P.
About two thirds of the world's P fertilizer is used on cereals, mostly on
wheat, rice, and maize (FAO, 2002), however it is challenging to assess the
"global" effectiveness of fertilizer P due to the absence of on-farm research.
Cereal yield improvements (AEP) of 20 to more than 50 kg grain/kg P applied
are normal for responsive soils. Most agricultural crops recover 20 to 30
percent of applied P during their growth under ideal conditions. The majority
of the leftover material settles in the soil and is gradually replenished by
succeeding crops over time, but even minor losses through runoff (dissolved
and particulate P) or leaching might have downstream effects off-site. REP
values for several field investigations on rice, wheat, and maize in Asia are
summarized in Table 6, the majority of which were conducted on soils with
low P fixation and under favorable climatic and management conditions. The
average REP ranged from 0.22 to 0.27 kg/kg P applied for all three crops.
However, REP varied greatly across all of these experiments, from zero to
almost 100% recovery. The most prevalent REP values (which account for
50% of all data) fell within the range of 0.10 to 0.35 kg/kg, which is likely
representative of most agricultural land worldwide.
Potassium
Global potassium flows are widely unbalanced because recoverable
natural K resources are concentrated at few locations (Sheldrick, 1985) and
potash use varies. North America, Western and Eastern Europe, and the
Middle East generate around 96% of the world's potash. Only a minor quantity
is produced in South America and Asia, and there is essentially no production
in Africa or Oceania. As a result, substantial quantities of potash fertilisers are
exported across the world to meet the need for this crucial macronutrient from

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crop production. Fortunately, potassium is safe for the environment and plays
a key role in raising agricultural output.
In many developing countries, K input-output budgets in agriculture are
highly negative. Nutrient audits have been conducted for several developing
countries (Sheldrick et al., 2002) and they mostly show a negative K balance.
Even though China's agricultural land has used more potassium over the past
20 years, its yearly K budget is still quite low, at roughly minus 60 kg K/ha.
Similar estimates predict yearly K losses of 20 to 40 kg K/ha for Indonesia
and India, and these losses have been continuously rising over the past 40
years.
REK values for several field investigations on rice, wheat, and maize in
Asia are summarized in Table 6. The average REK varied between 0.4 and 0.5
kg/kg K. REK is frequently in the 0.5 to 0.6 kg/kg range on soil with poor K
fixing capacity, with good management (high yield), and at relatively
moderate K rates. On-farm estimates of K usage efficiency are generally hard
to get.
Table 6: Average recovery efficiencies (kg/kg) of N, P and K from mineral fertilizers
in field trials with rice, wheat and maize in Asia. Values shown refer to
recommended fertilizer rates (rice, wheat and maize) or those currently applied by
farmers (rice).

Data set REN REP REK

Rice in S, E and SE Asia, farmers’ practice 0.33 0.24 0.38
Rice in S, E and SE Asia, site-specific management 0.43 0.25 0.44
Wheat in India 0.58 0.27 0.51
Wheat in China 0.45 0.22 0.47
Maize in China 0.50 0.24 0.44

Rice: 179 farmers’ fields in five countries, 1997-1998, N=314, (Witt and
Dobermann, 2004).
Wheat in India: field trials at 22 sites, 1970-1998. 120-26-50 kg/ha NPK
(Pathak et al., 2003).
Wheat and maize in China: field trials across China, 1985-1995 (Liu et
al., 2006).
Management strategies for increasing nutrient use efficiency
Nitrogen
• For a high PFPN, it's equally vital to understand and manage the N
supply from the soil and other local sources and to maximize

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 fertilizer efficiency (AEN = REN × PEN). Maximizing the first crop
recovery of N from mineral fertilizer or organic amendments (REN)
is particularly important due to the close link between yield and N
absorption and the fact that fertilizer-N losses are highest in the year
of application. Management should strive to obtain an AEN of 20-35
kg grain/kg N applied in current cereal production systems.
Typically, this requires a REN of 0.5-0.7 kg/kg.
• The amount of REN that may be attained depends on the crop's need
for nitrogen, the availability of nitrogen from local sources, and the
rate, time, kind, and application method of fertiliser. The difference
in REN between the minimum and maximum lines for any given level
of the index indicates additional elements, such as those that may be
influenced by better scheduling of N applications or other
management considerations. The highest levels of REN and profit are
not achievable by changing only one component using a particular
technology. Holistic management concepts are required that jointly
optimize-
1) The crop N sink for a specific environment.
2) The availability of soil and fertilizer-N for plant uptake at critical
growth stages.
• Numerous methods have additive impacts on how crops respond to
N, thus they must be used together:
1) Improved genetics (yield potential and abiotic/biotic stress
tolerance, N harvest index), knowledge of and exploitation of
seasonal yield potential, and the removal of additional barriers
to crop growth and internal N utilization are all ways to
maximize the crop N sink and the internal plant N utilization
(crop establishment, balanced nutrition, optimal water use,
control weeds, insects and diseases).
2) Better (site-specific) prescription algorithms, better timing of N
applications according to phenological stages, more efficient N
application methods, more efficient fertilizers (new N forms,
modified fertilizers, and inhibitors that lead to slow/controlled
release), and residue management for sustaining/increasing the
native soil N supply are all ways to manage soil and fertilizer-N
for better congruence with crop N uptake.
• Enhanced-efficiency Due to the fact that the information is
"embedded" in the product to be applied, N fertilizers have a

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 theoretical advantage over other more knowledge-intensive types of
N management. Embedded knowledge can result in high adoption
rates by farmers, as seen by experience with seeds, given that the
benefit/cost ratio is strong. The global effort to increase N usage
efficiency can thus benefit from improved fertilizer products, but the
relative relevance of these products will vary by area and cropping
system.
• The concepts for regulating N in organic farming systems are the
same as those for managing N from sources of mineral fertilizer.
• A combination of enhanced technology and local regulations that
encourage the deployment of such technologies is required to
increase N usage efficiency at the agricultural level. New
technologies must be resilient and lucrative, consistently deliver
benefits in N usage efficiency that are substantial enough, and take
up minimal additional time.
Phosphorus and potassium
Applying the current information in a useful way is the biggest obstacle
to increasing P and K usage efficiency at the farm level. Important factors
include:
• For every tonne of grain yield generated by cereals, 2-3 kg of P are
used, of which 70-80% are taken from the field with the grain.
Management should strive to attain an AEP of 30-50 kg grain/kg P
applied in current cereal production systems without significant P
fixation. An REP of 0.15 to 0.30 kg/kg is needed for this. It might be
challenging to set realistic target values for K usage efficiency
because of the diverse physiological roles that K plays in crop
production and crop K absorption. On soils with low levels of
accessible K reserves, feasible aims for cereals include AEK of 10-
20 kg grain/kg K applied and REK of 0.40-0.60 kg/kg.
• To optimize profit, prevent excessive buildup, and reduce the danger
of P losses on soils with moderate P and K levels and minimal
fixation, management must concentrate on balancing inputs and
outputs at the field and farm scales. In order to calculate the amount
of fertilizer needed as a function of the effective soil supply, net crop
removal, fertilizer recovery, and overall input-output balance, it is
necessary to have suitable prescription algorithms.
• Optimize crop management by removing other causes of inefficient
P or K consumption. Table 7 offers an illustration of this from a

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 lengthy rice experiment conducted in China. Rice showed a high
internal P efficiency (IEP = 590 kg/kg) when no P was administered
(NK treatment), suggesting P insufficiency. IEP = 345 kg/kg), but
because the system was K-deficient, it led to a subpar production gain
and an uneconomical soil P buildup. Adding P but bypassing K (NP
treatment) addressed the P shortfall. Due mostly to an increase in
REP, which also led to an increase in AEP and PFPP, balanced
fertilization (NPK) boosted yield.
• Many P and K recommendations in developing nations are based on
field studies that highlight the short-term response of crops to
fertilizer treatments. Although cereals' initial yield response to P or
K treatments is sometimes modest, over time, significant cumulative
yield gains can accumulate. Initial yield increases brought on by P or
K treatment in the case depicted in Figure 3 were negligible (0.5 t/ha).
However, when the amount of P and K in the soil that was accessible
to the plants decreased, production gains continued and grew with
time. A loss of 16.5 or 11 t/ha in grain output resulted from neglecting
P or K treatment, respectively.
• Plant portions that are vegetative contain the majority of the K that
plants absorb. The most crucial K management issue globally is to
improve internal, on-farm, and field recycling.
Table 7: Average rice yield (at 14% moisture), plant nutrient uptake, P use
efficiencies and cumulative P mass balance of eight consecutive rice crops grown at
Jinhua, China from 1997 to 2000 (Modified from Zhang et al., 2006).

Control NK NP NPK
Grain yield (t/ha) 2.7d 4.2c 4.9b 5.7a
N uptake (kg/ha) 37d 75c 83b 89a
P uptake (kg/ha) 6d 8c 15b 17a
K uptake (kg/ha) 43d 78b 58c 93a
IE of P (kg grain/kg P) 497b 590a 345c 352c
RE of fertilizer-P (kg P/kg P applied) 0.28b 0.35a
PE of fertilizer-P (kg grain/kg P) 157a 171a
AE of fertilizer-P (kg grain/kg P) 44b 60a
PFP of fertilizer-P (kg grain/kg P) 196b 226a
P input-output budget (kg P/ha/year) -12c -16d 21a 17b
Within each row, means followed by the same letter are not significantly different at
P<0.05 level.

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Summary
The focus will be on increasing N usage effectiveness and guaranteeing
more balanced fertilization, especially of K and micronutrients, in many
regions of Asia and South America. We intend to start a "Green Revolution"
in Sub-Saharan Africa, including the use of mineral fertilizers. A suitable
infrastructure and education are required for this.
Experience from a number of affluent nations has shown that encouraging
more advanced technology can buck the trend of diminishing N usage
efficiency. The highest possible levels of nutrient usage efficiency may be
attained in both rich and developing nations, according to research trials and
the world's top farmers. The difference between realistic aims and existing
levels of fertilizer usage efficiency, particularly for nitrogen, is still
considerable. The factors that control nutrient usage efficiency are well
understood. The examples in Table 8 provide as an illustration of how public
and private sector research and development have produced a variety of
technologies, tools, and regulatory initiatives for improving fertilizer usage
efficiency in real-world farming situations. Improvement is only possible by
implementing FBMPs at the field and farm scales, through systematic, site-
specific measures rather than promotion of general messages or "blanket"
solutions, as the use efficiencies of all major nutrients are driven by a
multitude of site-specific biophysical and socioeconomic factors. The latter
are crucial for promoting awareness and offering fundamental education, but
they require appropriate field-level diagnostic tools and management
strategies in order to be effective. The expansion of FBMPs requires
collaboration between the public and commercial sectors, as well as improved
support for recently made "greener fertilizing technology."
Three new challenges are emerging for public and private sector research,
the fertilizer industry and governments: climate change, bioenergy and
micronutrient malnutrition. Global climate will have profound but still little
understood influence on land use, crop yields, plant nutrition and a wide range
of other abiotic and biotic factors affecting the response to fertilizers (Lynch
and St. Clair, 2004; Pendall et al., 2004; Garrett et al., 2006; Long et al.,
2006; Pielke et al., 2007).
One of the most urgent Millennium Development Goals is malnutrition,
especially in South Asia and Sub-Saharan Africa. The new framework
(Graham et al., 2007) calls for attention first to balancing crop nutrition to
increase crop productivity, allowing sufficient staple to be produced on less
land so that the remaining land can be devoted to more nutrient-dense and

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nutrient-balancing crops. Once this is achieved, the additional requirements of
humans and animals for vitamins, selenium and iodine can be addressed.
Therefore, it is possible to improve nutrition through a mix of diverse meals,
enriched processed foods and water supplies, enriched crops with pro-vitamin
A and micronutrients through biofortification (breeding), or improved soil and
fertilizer management. Future efforts to improve micronutrient nutrition in the
poor countries will heavily include the fertilizer sector. There are several
methods for micronutrient enrichment ("fertification") of fertilisers, but
further research is required (IFA, 2005). In order to encourage the use of
enriched fertilisers in certain target locations, public policies must be devised.
Best management strategies for cultivating biofortified crops are little
understood. Many of those can only fully enrich genetically when the proper
FBMPs are used, including a minimum micronutrient supply.
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 Chapter - 6
Natural Farming: An Eco-agriculture

Authors
Komal Sharma
XXX
Minam Gamoh
XXX
Nisha Devi
XXX

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 Chapter - 6
Natural Farming: An Eco-agriculture
Komal Sharma, Minam Gamoh and Nisha Devi

Abstract
In India, the agriculture sector is important. It is indispensible for the
sustenance and growth of the Indian economy. More than half of the
population relies directly or indirectly on agriculture. In the mid 1960’s, the
introduction of green revolution was major breakthrough in the agricultural
sector and it was a transition from traditional agriculture. Nowadays, green
revolution’s exuberance is disappearing and it is becoming clear that the entire
technology has left a negative impact on the environment. Extensive use of
chemicals has led to many health and environmental issues. The biological
function of the soil was seriously harmed. Thus, need was felt to introduce a
more sustainable farming system which can ensure enhanced farm income and
reduces dependence on external inputs. Introduction of Natural Farming is a
viable and sustainable option to overcome the ill effects of chemical based
farming. Natural farming was first popularized by the Japanese scientist
Masanobu Fukuoka. In India, noted agriculturist Subhash Palekar has
introduced Zero budget natural farming (ZBNF) practice. ZBNF promises to
drastically cut production costs and enhance the farm income for the
comprehensive and long-term welfare of the farmers.
1) Ph.D. Scholar, Department of Agicultural Economics, Extension
Educaion & Rural Sociology, COA, CSKHPKV, Palampur,
Himachal Pradesh, 176062.
2) Ph.D. Scholar, Department of Agicultural Economics, Extension
Educaion & Rural Sociology, COA, CSKHPKV, Palampur,
Himachal Pradesh, 176062.
3) Assistant Professor, Department of Social Sciences, Dr. YS Parmar
University of Horticulture and Forestry, Nauni, Solan, Himachal
Pradesh, 173230.
Introduction
It hardly needs to be explained how important agriculture is to the
development of the Indian economy and society as a whole. Because it

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contributes significantly to both creation of jobs and the gross domestic
product, the agriculture sector continues to be an important segment of the
Indian economy (GDP). The Indian agriculture sector's resilience can be
demonstrated in the fact that despite the COVID-19 epidemic its output
performance was significant and recorded a decent growth rate of 3.4% in
2020-21 (Anonymous, 2020).
India has attained self-sufficiency in food grains production and has
emerged as major producer of many other agricultural products in the world.
This development has been achieved with modern technologies involving
extensive use of chemicals in the form of pesticides and fertilizers which has
led to many health and environmental issues. A severe agrarian crisis brought
on by the neo-liberalization of the Indian economy has rendered small-scale
farming an unprofitable vocation. Farmers are unable to access and pay for
privatised seeds, inputs, and market places. Indian farmers increasingly find
themselves trapped in a debt cycle as a result of their high cost of production,
high interest rates on credit, fluctuating agricultural market prices, rising input
costs dependent on fossil fuels, and the usage of proprietary seeds. When the
weather is bad, the issue grows worse. Indebtedness is contributing to an
increase in farmer suicides, which is contributing to the unfavourable
socioeconomic position that farmers currently face across the nation.
However, in consideration of novel prospects and challenges, it is felt that
Indian agriculture needs to undergo a transformational change. Agriculture is
currently confronted with three of the biggest issues of the twenty-first century
maintaining food and nutrition security, adapting and mitigating climate
change, making sustainable use of limited resources including water, energy,
and land. The Food and Agriculture Organization (FAO) advised all nations
to adopt agro-ecology in April 2018 in order to achieve the twin objectives of
ensuring global food security and environmental preservation.
Therefore, there is urgent need to make agriculture less capital intensive
so that farmer requires minimal credit to carry out agricultural production
activities. Growing consumer awareness about health and environmental
issues in agriculture has also led to need for production of chemical free food
and has emerged as a remunerative source of rural income generation. Organic
agriculture was initiated with this objective in mind but it failed to deliver the
desired results due to cumbersome, lengthy and costly production procedures.
Thus, need was felt to introduce a more sustainable farming system which can
ensure enhanced farm income and reduces dependence on external inputs.
Introduction of Natural Farming is a viable and sustainable option to
overcome the ill effects of chemical based farming as well as organic farming.

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Natural farming concept suggests that all macro and micronutrients required
by the crop/plants are present in soil but are in unavailable form. Plant use
nutrients from soil through mineralization and billions of microorganisms are
available in soil for this job. But continuous use of chemical fertilizers and
pesticides has reduced population of these useful microbes and need to be
rejuvenated. The use of cow dung and urine from indigenous Indian cows is
showing encouraging results as a good source of these bacteria.
One such low-input and climate-resilient agricultural method is known as
Zero Budget Natural Farming (ZBNF), which encourages farmers to use
inexpensive, locally obtained inputs instead of synthetic fertilisers and
commercial pesticides. Natural farming was first popularized by the Japanese
scientist and philosopher: Masanobu Fukuoka, who practiced it on his family
farm in the island of Shikoku. In India, Sh. Subhash Palekar introduced the
idea of zero budget natural farming which include a desi cow (Bos indicus) as
the focal point for input. Cow’s urine, dung, butter milk, jaggery, pulse flour
and other several homemade products serve as input. The basic principles of
natural farming are intercropping, agro-forestry, microbial inoculation and
increasing the activity of native soil biota. The inputs for natural farming are
homemade, low cost and have no negative ecological impact.
Subhash palekar natural farming
It is a naturistic method of farming in which desi cow products are used
by marginal and small farmers especially, as most of the farmers fall under
small and marginal farm category in India, having less than 1 hectare of land.
Essentially, there are four wheels of natural farming: 1) Beejamrit (Nectar for
Seeds), 2) Jeevamrit (Nectar of Microbes), 3) Acchadana (Mulching) and 4)
Whapasa (Soil moisture). For 30 acres, just one desi cow is needed, and
buffalo can also be employed in its place. However, Palekar advises that native
cow breeds have more microbes than their exotic counterparts and desi cows
are better adapted to Indian circumstances, less susceptible to disease and
easier to handle by resource-poor farmers. It is extreme form of LEISA (low
external input sustainable agriculture) that use the symbiotic relationship
between cow outputs and farm input. Cow dung is also said to attract
earthworms that are currently missing from our farms. It is a grass root level
movement started by Padmashri Subash Palekar (2006) in mid-1990’s which
spread through Karnataka, Andhra Pradesh, Punjab, Himachal Pradesh and
Kerala is planning to adopt it. India has about 82% small and marginal farmers
but till date there are only few viable options for small scale farmers which
cause a huge gap in earning of large and small-scale farmer. Therefore, Natural
farming system can be considered as a solution to small farmers’ income.

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The four pillars of ZNBF
1) Jeevamrit: A alternative for fertiliser, jeevamrit is prepared using
water (200 litres), cow dung (10 kg), cow urine (5-10 litres), pulse
flour (2 kg), jaggary (2 kg) and a small amount of farm soil. The
mixture is allowed to ferment for 48 hours and grow aerobic and
anaerobic microorganisms. Jaggery is believed to provide as a source
of nutrients for the soil's natural bacteria. In Jeevamrit, there is a very
high concentration of microbial load, which increases soil bio mass
even when it is supplied at a relatively low rate and improves soil
health. It also acts as a catalytic agent to promote soil microbial
activity and stimulate earthworm activity. Plant illnesses caused by
bacteria and fungi can be prevented by using Jeevamrit. According
to Palekar (2005), all of the nutrients needed for crop growth and
development are already present in the soil. Therefore, no additional
input is necessary. Instead, we must unlock the existing nutrients and
make them available using Jeevamrit. Jeevamrit is only necessary for
the initially three to four years following the system is self-
sustaining.
2) Beejamrita: It is a fermented cow-based substance used to guard
seeds against disease and soil contamination during the early stages
of germination. It is a microbial load and growth hormones rather
than a source of nutrients. It is a mixture comprised of water (20
litres), lime, 5 kg of cow dung, 5 litres of cow urine and a small
amount of soil. Beejamrita successfully protects young roots from
fungus, as well as from diseases transmitted by soil and seeds that
normally affect plants following the monsoon season. The same
ingredients as Jeevamrith are included in it, such as lime, soil, local
cow dung, which is a great biological fungicide and cow urine, which
acts as a high antimicrobial liquid. Cows and their products have
always held a special place in the lives of Indians, according to
ancient texts. According to Shubha et al. (2014), microbial
population increases when seeds get treated with Panchgavya or
Beejamrit.
3) Achhadana (Mulching): Mulching is a pragmatic way to alter the
crop-growing environment to increase crop quality and yield by
controlling soil temperature, preserving soil moisture and reducing
soil evaporation (Chakraborty et al. 2008). In Palekar's opinion, there
are three different kinds of mulching.

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 i) Soil mulch: This prevents tilling from destroying the topsoil
during farming. It increases soil aeration and water absorption.
Palekar advises abstaining from deep ploughing.
ii) Straw mulch: The term "straw material" typically refers to the
dried biomass waste from past crops and it can also include any
living thing's decomposing remains (plants, animals, etc). It is a
method used for improving soil fertility, by adding dry organic
material, which will break down and produce humus as a result
of microbial cultures' activation of the soil biota.
iii) Live mulch (symbiotic intercrops and mixed crops): In this
method in order to feed the soil and crops with all necessary
nutrients, different cropping patterns of monocotyledons and
dicotyledons must be established. For instance, legumes are
nitrogen-fixing plants that belong to the dicot group. Other
elements such as potash, phosphate and sulphur are supplied by
monocots like rice and wheat.
Other than these three methods, plastic mulching can also be used. It
leaves no chemical residue and is seen as a superior choice for disease control.
According to a review by Jalota et al. (2007), the usage of straw mulching
increased crop productivity in Punjab. Additionally, this method conserved
nitrogen fertiliser and irrigation water. Mulching has significant advantages
during the summer or kharif seasons and on soils with poor water retention.
According to Chakraborty et al. (2010), one of the main causes of air pollution
in the states of northern India is the burning of wheat and rice straw on the
Indo-Gangetic plains. Farmers are also given incentives by the central and
state governments to stop burning straw and hay, but these efforts don't appear
to be having any effect. Mulching has been observed to reduce water use by
3-11% and increase crop water efficiency by 25%. According to Mulumba and
Lal (2008), increasing mulch rates increased soil moisture retention at low
suctions by 70%, total porosity by 46% and accessible water capacity by 35%.
1) Whapasa (moisture): Plant roots don’t require a lot of water,
therefore in order to reduce the excessive dependency on irrigation
Natural farming method has been introduced. Palekar also believes
that water vapour is what roots require. He recommends limiting
irrigation and just irrigating at noon in alternate furrows since
whapasa is the state where both air and water molecules are present
in the soil.

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Other important principles of ZBNF
1) Intercropping: The name ‘Zero Budget’ primarily derived from the
concept of intercropping. It doesn't indicate that there won't be any
costs in farming, rather, it suggests that any costs will be
compensated by the income from intercrops, therefore making
farming free.
2) Contours and bunds: By creating contours and bunds rainwater can
be stored and maximise its effectiveness for various crops.
3) Local species of earthworms: In ZBNF local species of earthworms
play a vital role in improving the soil health. Hence it is advised to
boost organic matter in order to revive local deep soil earthworm.
4) Cow dung: In comparison to European cow breeds like Holstein, the
dung from the Bos indicus (humped cow) is the most beneficial and
contains the highest concentrations of microorganisms. The Indian
cow, which historically has been a part of Indian rural life, acts as the
focal point of the entire ZBNF methodology. These all principles
have also been depicted in Fig. 1.

Fig 1: SPNF principles

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What are the advantages of shifting to natural farming?
1) Small and marginal farmers who spend a lot of money on chemical
inputs will be benefited the most by taking up this type of farming.
2) Locally made stimulants can replace chemical fertilisers and
pesticides while retaining equivalent yields. This will lower
cultivation costs by 60 to 70%. Additionally, natural farming softens
the soil and improves the quality of the produce. As a result, farmers
may see a rise in their net income.
3) In ZBNF cost of cultivation is low as most of the inputs are already
available on the farms of farmers. Therefore, there is a huge
possibility of getting higher net returns as compared to chemical
farming.
4) Natural farming can liberate many farmers from predatory and
intertwined input and credit markets, therefore it will be helpful to
reduce the dependency on credit.
5) Every year government pays heavy funds on subsidies of the
fertilisers provided to the farmers which can be changed by adopting
ZBNF system. These expenses to the government funds can be
decreased by promoting natural farming.
6) Natural farming is more adaptable than organic farming since natural
farming is a progressive process whereas organic farming is more
about certification. As a result, small farmers find it simpler to
transfer to natural farming.
7) At the moment, people are compelled to buy food that contains
chemical residues. Although certified organic food is more
expensive, natural farming's inherent cost benefits can guarantee safe
food at reasonable pricing.
8) Climate change is mitigated by natural farming, which not only
reduces costs for farmers but also ensures a higher level of carbon
fixation in the soil.
9) The world's landscape may be replenished with water and can be
made greener by using natural farming techniques for land use
planning. It can also maintain the food's nutritional quality and
soil fertility.
10) Since natural farming doesn't use chemical fertilisers and pesticides,
it reduces marine pollution from land-based activities as well as

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 ocean acidification. Additionally, it aids in reducing pollution and
degradation of rivers and seas.
Challenges to natural farming in India
1) Some agricultural specialists believe it is naive to advocate for the
widespread use of natural farming methods since doing so could
seriously harm the valuable information and advantages that have
been gained from agricultural research and development over the past
70 years.
2) The crop protection market in India is estimated to be around Rs.
18,000 crores. Promoting natural solutions will put the entire
economic system at danger.
3) Natural farming can enhance soil health and lower insect infestation
rates but this does not mean that farmers can avoid using chemicals
when outbreaks occur.
4) Only 0.8% of the agricultural budget is allocated to India's National
Mission on Sustainable Agriculture by the Central Government.
(Anonymous, 2022).
What should be done to promote natural farming?
1) It's important to promote natural farming in a balanced way. As Sri
Lanka’s government immediately prohibited the use and import of
artificial fertilisers which caused a drastic decline in productivity and
a food shortfall.
2) The experience in Andhra Pradesh demonstrates that if farmers are
convinced and gradually migrate to natural farming, a process that
takes three to five years, a shift can be effective. Therefore, the
government must allow enough time and promote awareness
campaigns with real-world examples. To enhance farmer-to-farmer
capacity building for sustainable agriculture, civil society
organisations can be engaged.
3) Scientific study is required to support the practice of natural farming.
It's a wonderful start that the Indian Council of Agricultural Research
is developing a natural farming curriculum for undergraduate and
graduate courses.
Conclusion
The need for an alternative to existing farming system has been
demonstrated by literature that already exists on farmer emotion and well-

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being in India. Many farmers seek relief from the high input systems that
characterise Indian agriculture because they can have detrimental effects on
the economy, the environment and society. The severe effects of traditional
farming include farmer suicides, emigration and soil erosion, causing a danger
to food security and rural India's well-being. ZBNF addresses these problems
by offering an alternative to traditional farming methods and giving farmers
more freedom from debt and autonomy. Although some early indicators of
ZBNF's impact on finances and yield are favourable but long-term and system-
wide data must be gathered to confirm these effects. Furthermore, there are
still gaps in the literature regarding how ZBNF alters foods capes, ecological
health and farmer well-being. The effective scaling up of ZBNF may also be
benefited from additional governmental support which will include financing
for research, formal certification systems for ZBNF crops, reform of tenant
contracts and more targeted resource allocation to ease the financial burden of
the transition period.
While the majority of the preliminary findings on ZBNF are encouraging,
there is a significant gap in the published literature that has to be filled in order
to make well-informed policy choices on state-wide adoption of ZBNF. Crop
production appears to vary depending on region, research methodology and
sample size making it challenging to generalise the results. To understand how
ZBNF affects yield, more thorough study is required, including multi-year
studies that start taking farm scale and growing conditions into account. There
are no peer-reviewed researches that look at how ZBNF alters soil
composition, especially over a lengthy period of time. Realizing this is
essential for determining ZBNF's long-term viability and sustainability in
India. To support effective decisions regarding the state-wide scaling up of
ZBNF, long-term, peer-reviewed studies that address knowledge gaps on
climate effects, gendered impacts, youth engagement, migration impacts, and
system-wide productivity and profitability are essential. Without this
information, it would be very challenging to decide whether ZBNF should be
implemented throughout the entire state or only aimed at particular farmers in
specific regions.
This is possibly the most significant factor going forward because
preliminary data indicate that ZBNF won't be a universal solution that
produces the same results for everyone. The next crucial step in implementing
these practices across the entire state is to identify the elements that make a
ZBNF system effective. These factors as well as a larger understanding of how
ZBNF may affect the profile of crops grown in different agro-climatic zones
should be addressed in order to make sound policy decisions that benefit
farmers and promote food security.

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Future prospects of study
So far, an assessment of "Zero Budget Natural Farming" has clearly
demonstrated its remarkable viability for various agro-climatic situations,
crops, and farmer categories. It has gained popularity among farmers since it
perfectly complements their lifestyle, which is reliant on the soil, greenery and
cattle. However, it must be observed over a period of time in order to see
whether it can offer consistent returns year after year. Without a doubt,
Paleker's zero budget natural farming has had a lasting impression on Indian
agriculture. This method has substantially benefitted lakhs of small-scale
farmers throughout the nation, and as time goes on, this number will rise
dramatically.
References
1. Anonymous. 2020. Economic survey 2020-21. Department of Economic
Affairs, New Delhi.
2. Anonymous. 2022. Natural Farming- processes, advantage and
challenges.
[Link]
challenges/
3. Palekar, S. (2006). The principles of spiritual farming II. 2nd ed. Amravati:
Zero Budget Natural Farming Research, Development & Extension
Movement, Amravati, Maharashtra, India.
4. Shubha S., Devakumar N., Rao G.G.E., & Gowda, S.B. (2014). Effect of
Seed treatment, Panchagavya application and Organic Farming Systems
on Soil microbial population, Growth and Yield of Maize. Proceedings of
the 4th ISOFAR Scientific Conference. ‘Building Organic Bridges’, at
the Organic World Congress 2014, 13-15 Oct. Istanbul, Turkey
5. Palekar, S. (2005). The philosophy of spiritual farming I. 2nd ed.
Amravati: Zero Budget NaturalFarming Research, Development &
Extension Movement, Amravati, Maharashtra, India.
6. Chakraborty, D., Nagarajan, S., Aggarwal, P., Gupta, V.K., Tomar, R.K.,
Garg, R.N., Sahoo, R.N., Sarkar, A., Chopra, U.K., Sarma, K.S.S., and
Kalra, N. (2008). Effect of mulching on soil and plant water status and
the growth and yield of wheat (Triticum aestivum L.) in a semi-arid
environment. Agriculture Water Management, 95:1323- 1334.
7. Jalota, S.K., Khera, R., Arora, V.K. & Beri, V. (2007). Benefits of straw
mulching in crop Production: A review. Journal of Research, Punjab
agricultural University, 44(2): 104-107.

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8. Chakraborty, D., Garg, R.N., Tomar, R.K., Singha, R., Sharma, S.K.,
Singh, R.K., Trivedi, S.M., Mittal, R.B., Sharma, P.K., & Kamblea, K.H.
(2010). Synthetic and organic mulching and nitrogen effect on winter
wheat (Triticum aestivum L.) in a semi-arid environment. Agricultural
Water Management, 97: 738-748.
9. Mulumba, L.N., & Lal, R. (2008). Mulching effects on selected soil
physical properties. Soil & Tillage Research, 98: 106-111.

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Page | 104
 Chapter - 7
Rhizosphere: Role and Importance for Soil
Health Management

Authors
Varun Parmar
Ph.D. Scholar, Department of Soil Science, CSK HPKV,
Palampur, Himachal Pradesh, India
Naveen Datt
Principal Scientist, Soil Microbiology, Department of Soil
Science, CSK HPKV, Palampur, Himachal Pradesh, India
Rushali Katoch
Ph.D. Scholar, Department of Soil Science, CSK HPKV,
Palampur, Himachal Pradesh, India

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Page | 106
 Chapter - 7
Rhizosphere: Role and Importance for Soil Health
Management
Varun Parmar, Naveen Datt and Rushali Katoch

Abstract
With the rising human population, predominant reliance on an
agricultural sector to supply food is becoming unprecedent. It is an undisputed
fact that in order to meet the food security, the farming community is largely
dependent on chemical inputs. Over the last few decades, the injudicious use
of chemical inputs viz. fertilizers, pesticides, herbicides have resulted in
serious environmental concerns. Additionally, accelerated industrialisation
and other anthropogenic activities such as uncontrolled use of agrochemicals
and the dumping of sewage sludge have significantly degraded soil health by
contaminating soils with numerous pollutants like heavy metals and organic
pollutants. Soil is a primary natural resource that maintains ecosystem
functioning, water balance, and supports plant growth. Soil is also a habitat
for diverse microflora that is very crucial to soil sustaining activities. The
rhizosphere is undoubtedly the most complex microhabitat, comprised of an
integrated network of plant roots, soil, and a diverse consortium of bacteria,
fungi, eukaryotes, and archaea. The rhizosphere conditions have a direct
impact on crop growth and yield. Nutrient-rich rhizosphere environments
stimulate plant growth and yield. Although the importance of the rhizosphere
microbiome for plant growth has been widely recognized, but no knowledge
exists for the vast majority of rhizosphere microorganisms. To enhance plant
growth and health, it is essential to know which microorganism is present in
the rhizosphere microbiome and what ecosystem function they perform.
PGPM’s (Plant growth promoting microorganisms) include bacteria,
actinobacteria, fungi and others which live in a mutualistic relationship along
with plants (symbiotic/associative/free living/ endophytic) and make the
nutrients available to plants, play a major role in organic matter
decomposition, produce phytohormones, phytoremediation, help the plant to
withstand abiotic stress and act as biocontrol agents for plant pests and
diseases. Although PGPM’s are already present in soil, still there is a dire need

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of elevating their strength and activity through direct application, following
different agronomic interventions and utilising modern biotechnological and
molecular techniques to identify, classify, characterize and ultimately
fabricate these rhizosphere workers in order to sustain soil health as well as
crop productivity.
Keywords: Rhizosphere, soil health quorum sensing, rhizosphere
engineering, siderophores, microbiome, phytohormones
1. Introduction
After independence, besides recovering from the wounds of partition,
India had to still ensure food security to millions of people. The “green
revolution” in 1960’s and 70’s came to the rescue that employed use of input
responsive high yielding variety seeds, improved irrigation facilities,
mechanized farm tools and intense application of chemical fertilizers and
pesticides. Undoubtedly, green revolution not only boosted India’s food grain
production but also transformed it from being a food importer nation to a
substantial exporter. Notwithstanding, with all the efforts, food insecurity is
still a chronic issue that is likely to worsen as the human population is expected
to be more than 9 billion by 2050 (Kumar and Dubey, 2020).
Moreover, amazingly increased human population exerts a continuous
pressure on farmers to enhance crop productivity and to achieve this target,
injudicious use of chemical inputs is being practiced in agriculture system.
However, to keep the pace with increasing population, crop production must
increase but not at the expense of environment and ecology. Frequent use of
chemical inputs resulted in considerable decline in microbial activity,
nutritional imbalance and a drop in the population of beneficial microbes in
the soil vis-à-vis deteriorating soil health. In order to sustain the dynamic
equilibrium of the soil ecosystem, a healthy soil has a "harmonious social
system" with good foundation, functional state and buffering performance.
Therefore, maintenance of soil health is essential for developing healthy
ecosystem and sustainable agricultural production.
Soil is not only estimated to contain one-third of all living organisms but
is also the site for myriad of interactions occurring in it that regulates the
activity of the organisms responsible for ecosystem functioning and evolution
thereby making it the most complex habitat on earth. Rhizosphere offers
enormous potential for agricultural benefit in terms of ensuring food security,
crop production sustainability and making agricultural systems climate change
resilient.

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2. Rhizosphere and organisms present
Rhizosphere is the region in the soil in the vicinity of roots which is
governed by plant roots. Apart from the beneficial microbes, plant pathogenic
microorganisms also colonize the rhizosphere striving to break through the
protective microbial shield and to overcome the innate plant defence
mechanisms in order to cause disease. Moreover, a third group of
microorganisms found in the rhizosphere are human pathogenic bacteria,
which can be carried on or in plant tissue and may cause disease when
introduced into the humans (Mendes et al. 2013). To unleash the full potential
of beneficial rhizosphere microbes in order to enhance plant growth and
maintain soil health, it is first essential to know which microbe is present in
the rhizosphere microbiome and what specific role they are playing there. In
general, the rhizosphere region is less diverse when compared with the bulk
soil. Among the host of other factors microbial diversity in the rhizosphere
depends upon type of plant species, its root structure, physical and chemical
properties of soil.
3. Role of rhizosphere microorganisms in soil health management
3.1 Nitrogen fixation
Despite being the most abundant gas in the atmosphere (78.7%), Nitrogen
(N) in its dinitrogen (N2) form is not accessible to plants directly, rather it is
taken in the form for ammonia and nitrates through their roots. The ammonium
form is directly assimilated into amino acids while the nitrate has to be
converted into ammonium before it can be used (Beeckman et al., 2018). N
constitutes about 2% of the total dry matter of a plant and is one of the most
important nutrients required for plant growth. Plants need N for the synthesis
of nucleic acid, proteins, and enzymes (Bano and Iqbal, 2016) while its
deficiency leads to yellowing of older leaves, stunted growth, smaller leaves,
shorter internodes and reduced protein content.
Biological N fixation is perhaps the second most important biological
process after photosynthesis which involves conversion of dinitrogen to
ammonia facilitated by symbiotic and free-living group of prokaryotes.
During the last 45 years, the demand for nitrogen fertilizers has enhanced from
12 to 107 Tg year-1 and is expected to increase to 111.5 Tg year-1 in the year
2022 (FAO, 2019). With such an enormous fertilizer demand, the likelihood
of soil degradation cannot be disregarded. Nonetheless, biological nitrogen
fixation can be an alternate source for meeting plant N requirement. The input
of nitrogen into soil through BNF ranges from 0 to 60 kg ha-1 year-
1(Reghuvaran et al., 2012) with an estimated contribution of 175 million

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metric tons annually covering 70% of all annual fixed nitrogen on the Earth
(Lodewyckx et al., 2002). Rhizobium and Bradyrhizobium are the main genera
of bacteria that fix atmospheric nitrogen symbiotically. The process of
nitrogen fixation begins with the infection of root hair cells by rhizobia which
is facilitated by the Release of certain signaling compounds i.e. flavonoids
such as flavonones and betains by legumes in their root exudate (Gage 2004).
Rhizobial NodD transcriptional activator proteins then recognize flavonoids
and resultingly induce the Transcription of nod genes. The nod genes encode
approximately 25 proteins that are involved in the synthesis and export of a
lipochitooligosaccharide signaling molecule called Nod factor (NF) (Gage
2004). Plasma membrane of legumes consists of NF receptors such as kinase
that contain chitooligosaccharide binding sites (Limpens et al. 2005; Radutoiu
et al. 2007) which recognizes nod factors and thereby resulting in an increase
in concentration of Ca in nucleus and ultimately inducing physiological
responses by the legume such as root hair curling. Resultingly, the rhizobia
are trapped within the curl and continue to divide forming colonies called
infection foci. The rhizobial division is followed by the plant cell wall
degradation which begins at the site of contact between bacteria in the
infection foci and the plant cell wall. The rigorously dividing rhizobia are then
taken up into the root via infection thread and elongates into Bacteroides
which catalyzes nitrogen fixation with the aid of nitrogenase enzyme. The
nitrogenase enzyme consist of MoFe protein complex which binds to diatomic
nitrogen gas and transfer electrons that allow the reduction of the triple bond
in nitrogen to ammonia. Reduced nitrogen (NH3) is assimilated by the plant
into either purine derivatives or amides and is translocated from the nodules
via the xylem (Cordoba et al. 2003; Barsch et al. 2006).
3.2 Phosphorus solubilization
Being a 2nd most important macronutrient for plants, phosphorus is
essential for all key physiological and biochemical processes including cell
division, photosynthesis, respiration, the development of the root system, and
the creation of macromolecules (Sharma et al., 2012). It also significantly
contributes to the stem strength, crop maturity and quality improvement in
many fruits, vegetables, and cereal crops (Sagervanshi et al., 2012). At cellular
level, all energy transfers in the cell are critically dependent on phosphorus.
Although phosphorus is widely distributed in the soil in both inorganic
and organic forms, but a small fraction (0.1%) is present in the soluble (plant-
available) form (HPO42- or H2PO4-). About 20-80 per cent of overall
phosphorus pool is organic phosphorus, which is found in soil humus and
serves as a significant immobilised phosphorus reservoir. Most of the organic

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phosphorus is found as inositol phosphate (10-50%), sugar phosphates,
phospholipids (1-5%), nucleotides (0.2-2.5%), phosphoproteins, and
phosphonates. The inorganic form of phosphate sequestered in soil either by
adsorption to the soil mineral surfaces or through precipitation. In order to
sustain crop production, large amounts of chemical phosphate fertilizers are
added to the soil since cultivable soils lack sufficient available phosphorus but
only a small fraction of applied fertilizer is absorbed by the plant while the
remaining is transformed into an insoluble form that is unavailable to the plant
(Cabeza et al., 2019).
While the insoluble organic phosphorus is transformed into soluble form
through mineralization, the inaccessible inorganic phosphorus is hydrolysed
through solubilization process. Solubilization process is mediated by several
microorganisms viz. bacteria, fungi and actinomycetes by producing different
type of organic acids and enzymes. Organic acids chelate with the cations
present on mineral surface of soil thus block the phosphate adsorption sites
and as a result increase available phosphorus in soil. Bacillus, Pseudomonas,
Flavobacterium, Enterobacter, Klebsiella, Serratia and other bacterial genera
releases phosphatases and phytases enzymes that have higher affinity for
organic phosphorus to convert them into soluble form.
3.3 Phytohormones production
Phytohormones are important regulators of plant behaviour and are
important for physiological and developmental processes. Plant hormones
have historically been categorised into five groups: auxin, gibberellins,
cytokinins, abscisic acid and ethylene (Ole´nska et al, 2020). Numerous
rhizosphere-dwelling bacteria are capable of producing phytohormones,
which aid in the growth and development of plants.
a) Auxin
Auxin is the most important phytohormone that regulates almost every
aspect of plant development. The most prevalent, well-characterized auxin
produced by bacteria and plants is indole-acetic acid (IAA), which plays an
important role in seed germination, apical dominance, root development, cell
division and cell differentiation. Additionally, it also helps in metabolites
biosynthesis, stress resistance and photosynthesis. IAA synthesis by
microorganisms generally uses three pathways: 1) Bacterial genera viz.
Erwinia, Agrobacterium, Pseudomonas, Azospirillum, Bradyrhizobium,
Enterobacter, Klebsiella and Rhizobium produces IAA via the intermediates
indole-3-pyruvic acid and indole-3-acetic aldehyde, 2) Azospirillum and
Pseudomonas has been reported to produce IAA via tryptamine and indole-3-

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acetic aldehyde and 3) In Agrobacterium, Erwinia and Pseudomonas strains,
indole-3-acetamide (IAM) production is used to synthesise IAA (Tahir and
Sarwar, 2013). Raheem et al., 2018 suggested to use auxin producing PGPRs
for rhizosphere engineering in drylands as they showed positive effect on
growth and yield parameters of wheat grown under drought conditions.
b) Gibberellin
Gibberellins (GAs) are plant hormones that are involved in almost all
phases of plant growth and development, such as embryogenesis, stem
elongation, blooming, leaf expansion and fruit ripening (Binenbaum et al.,
2018). Natural GAs are conjugates of b-D-glucose however, they can also
exist in free and bound states. Rhizobium meliloti was the first bacterial species
to have gibberellins GA1, GA4, GA9 and GA20 (Atzorn et al., 1988) and apart
from this, bacterial genera such as Acetobacter, Azospirillum, Herbaspirillum,
Bacillus and Pseudomonas have also been reported to produce GAs
(Maheshwari et al., 2015). According to Kang et al. (2014), Leifsonia soli, a
rhizobacterium demonstrated the ability to produce GAs and promoted the
growth of tomato, cucumber and radish plants.
c) Cytokinin
Cytokinin, an important group of plant hormone is involved in number of
physiological and developmental activities, such as apical dominance, seed
germination, nodule formation, flower and fruit development, root elongation,
vascular development and interactions between plants and pathogens.
Cytokinin is produced and released by bacteria in the rhizosphere,
subsequently increasing the concentration in the soil solution and ultimately
stimulate the growth of plants. Furthermore. The PGPRs releasing cytokinin
are also involved in regulating defence mechanism of the host plant against
pathogens. Cytokinin produced by Pseudomonas fluorescens successfully
managed the infection caused by Pseudomonas syringae while maintaining
the tissue integrity and yield in Arabidopsis (Grosskinsky et al., 2016).
d) Abscisic acid
Similar to other phytohormones, abscisic acid (ABA) is recognised to
play a significant function in plants by enhancing stress responses and
adaptation. ABA has also been found to affect root development and water
content under drought stress conditions (Cutler et al., 2010). However, sudden
rise in ABA concentrations during stress exposures can lead to growth
retardation and can also modulate tolerance responses against various stresses.
Mora-Herrera and Lopez-Delgado (2007) found that ABA application
improved stress tolerance in Solanum tuberosum by lowering the free radicals

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through considerable increases in the activity of the antioxidant enzyme
peroxidase.
e) Ethylene
Certain PGPRs produces a vital enzyme, 1-aminocyclopropane-1-
carboxylate (ACC) deaminase, which regulates ethylene production by
metabolizing ACC into α-ketobutyrate and ammonia. By lowering stress-
induced ethylene production, inoculation with these PGPRs sustain plant
growth and development under stressed conditions. Furthermore, these
rhizospheric bacteria have the capacity to enhance ACC outflow from roots.
A substantial amount of ACC is secreted by the plant to maintain homeostasis
both inside and outside the roots. Consequently, the growth of ACC deaminase
producing beneficial microbes in the rhizosphere is accelerated, which lowers
the concentration of ACC in the plant's roots, resulting in less ethylene
production and improved plant growth (Gamalero and Glick, 2015).
3.4 Biocontrol agents
To ensure sustainable plant development and soil fertility, the use of
PGPR is a practical and eco-friendly strategy. This strategy advocates the
management of extensive PGPRs to prevent the spread of pathogens. Various
mechanisms are involved to kill the phytopathogens such as production of
siderophores, antibiosis and lytic enzymes etc. Siderophores are low
molecular weight iron-chelating compounds having great potential to provide
iron to the cell or tissues of a plant (Hider and Kong, 2010). PGPR synthesized
siderophores exhibit a strong affinity for rhizospheric Fe3+ and retain almost
all of the free iron thus enhancing iron absorption by plants and preventing the
spread of harmful microorganisms in the vicinity of roots. Pseudomonas sp.
boosted plant development and slowed the spread of disease in green gram by
producing siderophores (Sahu and Sindhu, 2011).
Antibiotic production by rhizobacterial strains is also one of the most
effective mechanisms against pathogenic microorganisms. Phloroglucinols,
phenazines, pyoluteorin, cyclic lipopeptides and pyrrolnitrin are a few of the
six distinct antibiotic groups that have been identified till now. The majority
of these antibiotics are linked to the biological control of diverse root
infections. Bacillus and Pseudomonas species produced lipopeptide bio-
surfactants, which have been evaluated in the biological control of several
pathogenic microorganisms (Ali et al., 2020).
Secretion of lytic enzymes is essential characteristic for biological control
of pathogenic microbes in rhizosphere. PGPR produces different lytic
enzymes viz. dehydrogenase, chitinases, phosphatases, lipases, proteases etc.

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which plays a significant role in stimulating plant growth mainly by protecting
plants against several pathogenic fungi such as Sclerotium rolfsii, Fusarium
oxysporum, Pythium ultimum and Rhizoctonia solani (Yadav et al., 2016).
Disrupting structural stability and integrity of cell walls of target pathogens,
is the main mechanism for its action.
3.5 Iron chelation
Iron (Fe) is an essential element for the growth of almost all living
organisms because it acts as a catalyst in enzymatic processes, oxygen
metabolism, electron transfer, and DNA and RNA synthesis (Aguado-
Santacruz et al., 2012). Iron is not easily available to plants particularly in
calcareous soils therefore plants employ different strategies to take up the
nutrient; one such strategy is microbe mediated siderophore production which
is a high affinity iron chelator that enables the plant to acquire it even at low
concentrations. Siderophores, through ligand exchange binds with ferric ion
(Fe3+) thereby making a soluble complex in soil. The siderophore receptors
present at the cell membrane of plant roots recognize the Fe-siderophore
complex and are taken up by the cell (Singh 2020).
Several bacteria such as Agrobacterium tumefaciens, Azospirillum,
Azotobacter, Bacillus, E. coli, Enterobacter, Mycobacterium, Neisseria
gonorrhoeae, Paracoccus denitrificans, P. fluorescens, Rhizobium meliloti,
Serratia, and Streptomyces, etc. have been reported to synthesize siderophores
(Ghosh et al., 2020; Singh, 2020). Many researchers have reported to increase
the productivity of field crops with application of siderophore secreting
microbes by increasing iron availability in soil (Karnwal, 2017, Gull and
Hafeez, 2012, Sah et al., 2017 and Khalid et al., 2015). Ipek et al., 2017
reported an improvement in iron content of plant root and leaves and enhanced
the activity of ferric chelate-reductase (FC-R) enzyme in pear grown under
calcareous soil conditions by applying PGPR strains Agrobacterium sp.,
Alcaligenes sp., Bacillus sp. Pantoea sp., and Staphylococcus sp.
3.6 Abiotic stress tolerance
Numerous studies have reported rhizobacteria mediated tolerance in
plants against various abiotic stresses (Rajput et al., 2018). However, the
levels of tolerance depend on soil microbes' capacity to activate plant systems
for the expression of stress-responsive EPS, transcription factors, proline
synthesis and biomass stabilisation. Recent research has indicated that
phytohormones generated by microorganisms associated with roots may be
considered as essential metabolic engineering targets for promoting host
tolerance to abiotic stresses particularly under drought and salt stress.

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A) Drought stress
PGPR exhibit a variety of tolerance and adaptability strategies to deal
with decreased water potential, including osmolyte accumulation, thickening
of walls, spore formation and EPS synthesis (Kour et al., 2019). Proline,
trehalose and betaine accumulation aids bacteria in maintaining membrane
permeability, integrity and functional proteins during drought stress (Asghari
et al., 2020). For instance, compared to uninoculated plants, maize seedlings
inoculated with Azospirillum brasilense demonstrated improved relative water
content. Inoculating plants with bacteria that can withstand drought prevents
a significant loss in plant water potential and raises proline levels in the leaves
and roots, which further promotes plant growth. In presence of water, impact
of rhizobacteria is more considerable (75%) (Casanovas et al., 2002).
B) Salt stress
Halo-tolerant rhizobacterial strains can survive in high saline soils and
possess certain qualities that help plants to flourish in a saline environment
(Abbas et al., 2019). Numerous studies have documented the positive effect
of PGPR on a variety of plants against the negative effects of high salinity
using a variety of mechanisms, including biofilm formation, osmolyte
accumulation, maintaining turgor pressure of salt-stressed cells etc. (Nawaz et
al., 2020). The isolated strains of Bacillus subtilis and Bacillus pumilus from
saline soil displayed a variety of PGPR characteristics, including phosphate
solubilization and IAA synthesis, as well as tolerance to salt stress
(Damodaran et al., 2013). The use of Pseudomonas putida in the chickpea
plant demonstrated thermotolerance through the development of a thick
biofilm and the production of transcription factors linked to stress (Kumar et
al., 2016).
3.7 Heavy metal toxicity
Heavy metal (HM) pollution is a major environmental problem which
reduces crop production and food quality due to excessive application of
agricultural inputs like fertilizers, pesticides and have resulted in the heavy
metal contamination of soils (Su, 2014). Most of the pesticides are organic
compounds, and a few are inorganic compounds or pure minerals, and some
pesticides contain Hg, As, Cu, Zn, and other heavy metals (Arao et al. 2010).
Unlike organic contaminants, metals are not degradable and thus remain in the
environment for long periods of time; when present at high concentrations,
metals can negatively affect plant metabolism (Ferraz et al. 2012). Biosorption
and bioaccumulation are the two most common mechanisms utilised by
microbes to render soils free from heavy metal contamination. The term

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biosorption is collectively used to refer adsorption and absorption by living or
dead biomass whereas bioaccumulation involves taking up of a substance inti
the matrix by living cells only. The bacterial cell wall contains various
functional groups such as phenols, carboxylic acid, etc which contain negative
charge. Heavy metal cation being positively charged gets electrostatically
attracted to the oppositely charged functional group thereby getting adsorbed
on the cell wall matrix. Once the heavy metal cation associates itself with the
bacterial cell wall, it is then transported inside the bacterial cell with the help
of specialized protein-based carriers. Consequently, the cation is either utilised
by the microbe in its metabolism or is reduced into non-toxic form (Saha et
al. 2017).
4. Conclusion
Since the beginning of the civilisation, humans have evolved and learned
to harness the natural resources as per their needs. But over the centuries
human population has escalated tremendously resulting in an undue pressure
on existing natural resources including soil. Rapid industrialization coupled
with over dependence on pesticides and fertilizers and poor management of
natural resources has forged an auxiliary hurdle in the path of sustainability.
Sustainable crop production is the need of the hour. Numerous technologies
have been delineated by the researchers to cater and opt different techniques
that are not only environment friendly but also economically viable. The
rhizosphere is highly dynamic and nutrient dense region surrounding the
immediate surface of roots. Various species-specific secretions from plant
roots makes it highly dense but less diverse when compared with the bulk soil.
Rhizosphere not only influence soil health but also help the plants to withstand
biotic as well abiotic stress. With their distinctive potential to solubilize and
sequester insoluble form of different nutrients, it is imperative that the
rhizosphere microbes should be extensively studied and implemented in
modern agriculture.
5. Future thrust
The rhizosphere microbiome harbours heterogenous group of PGPR’S
which possess significant potential to alter the rhizosphere. Due to the
complex nature of interactions between soil, microbes and plants and the
limitation of studying only the culturable microbes, it becomes extremely
important to find the best technology to identify and characterize important
PGPR’s. The knowledge of these plant-microbe interactions can help in
developing new sustainable, eco-friendly, and economically viable systems
for agriculture (Kumar et al., 2015; Rakshit et al., 2015; Naqqash et al., 2016).
Rhizosphere is dynamic in nature and is very much influenced by its

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components. Root and soil are complex microbial habitats harbouring diverse
microbial consortia. Understanding, predicting and controlling the structure
and function of the rhizosphere will allow us to harness plant-microbe
interactions and its activities to increase or restore plant ecosystem
productivity, improve plant responses to a wide range of environmental
alterations in the function and mitigate effects of climate change by designing
ecosystems. In the future advanced technologies such as quorum sensing (QS),
rhizosphere engineering and metagenomics must be utilized to not only
identify the specific PGPR but also understand the complex mechanism they
utilise and subsequently produce effective and eco-friendly strains.
QS is the inter-cellular communication mechanism in which microbial
gene expression is coupled with the concentration of bacterial cells, is
regulated through the diffusion of particular signalling components such as
acyl homoserine lactone AHLs (Awan et al., 2011). Rhizosphere engineering
is the process in which manipulation of rhizosphere microorganisms is done
for obtaining desired trait(s). This is basically done by the alteration in root
exudation pattern of plant, which can be achieved through genetic
manipulation in the host plant and by natural induction through interventions
in soil. Metagenomics is the study of the structure and function of entire
nucleotide sequences isolated and analysed from all the organisms (typically
microbes) in a bulk sample. Metagenomics provides access to the functional
gene composition of microbial communities and thus gives a much broader
description than phylogenetic surveys, which are often based only on the
diversity of one gene, for instance the 16S rRNA gene. Coupling traditional
approaches with advanced next generation sequencing techniques to assess
organismal or community ecology and physiology will bring new insights to
understand microbial life in the rhizosphere.
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 Chapter - 8
CRISPR - Genome Editing Tools for Genetic
Improvement in Agriculture

Authors
Prachi Mahla
Ph.D. Research Scholar, Department of Genetics and Plant
Breeding, RCA, MPUAT, Udaipur, Rajasthan, India
Sheenam Bhateja
Ph.D. Research Scholar, Department of Entomology, RCA,
MPUAT, Udaipur, Rajasthan, India
Himansuman
Ph.D. Research Scholar, Department of Genetics and Plant
Breeding, RCA, MPUAT, Udaipur, Rajasthan, India

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Page | 124
 Chapter - 8
CRISPR - Genome Editing Tools for Genetic Improvement
in Agriculture
Prachi Mahla, Sheenam Bhateja and Himansuman

Abstract
The demand for food crops has significantly increased as a result of the
rapid growth of the world population. Breeders worked hard over a long period
of time to boost crop yield and enhance quality using conventional breeding
techniques. The conventional breeding techniques will be unable to fulfil the
future food demand. To combat this future threat breeders are using several
modern molecular technologies for rapid crop improvement. A new popular
gene editing tool CRISPR/Cas9 has been widely used for rapid crop
improvement. Due to its simplicity and efficiency it is widely used by the
researchers for gene editing in both plant and animal genomes. CRISPR/Cas9
technology uses a Cas9 protein and a guide RNA for cleaving a genome at e
specific loci. This book chapter summarizes the mechanism of CRISPR
technology, its applications in crop improvement and future prospects.
Keywords: CRISPR/Cas9, gene editing, crop improvement, crop yield
1. Introduction
The rapidly increasing world population has resulted in a significant
increase in the demand for food crops. Traditional crop breeding approaches,
on the other hand, are labour intensive and time consuming. As a result,
available crop types and current agronomic practices will be unable to meet
future global food demand. Furthermore, rising crises that result in lower crop
yields and quality are aggravated by gradual climate changes and the loss of
natural genetic resources. In this perspective, the development and
deployment of gene-editing tools that may rapidly enhance crop qualities
through precise genetic modifications is a promising option for accelerating
germplasm resource improvement and the introduction of novel varieties to
meet the difficulties of feeding the world [1]. Researchers now have powerful
tools for decoding gene functions and enhancing plant traits owing to the
recent emergence of gene-editing technologies involving zinc finger nucleases

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(ZFNs), transcription activator-like effector nucleases (TALEN), and
clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-
associated protein (CRISPR/Cas) systems. Using artificially constructed
nucleases, gene-editing technologies allow for the change of a specific
genomic location (e.g., gene knockout/single-base editing and guided editing)
[2]
. Because of its simplicity and convenience, the CRISPR/Cas system has
been widely utilised to modify plant genomes and create mutants. It is
increasingly being used to change the characteristics of various plants,
including vital crops, as well as to produce new germplasm resources. The
CRISPR/Cas9 system, which is widely used for genome editing, involves the
RNA-guided DNA endonuclease Cas9 cleaving DNA. The optimization of
this approach makes it easier to modify target genes in an efficient and
accurate manner, hence speeding plant breeding. As a result, CRISPR/Cas9
has evolved into the most frequently used and advanced gene-editing system
[3]
.
This chapter introduces the CRISPR/Cas9 genome editing system by
describing the progress in the associated research, the underlying mechanism,
the related technology, and it’s utility for crop breeding. Furthermore, the
limitations of the CRISPR/Cas9 system and how they may be overcome are
discussed. This chapter provides researchers with critical information relevant
for applying CRISPR/Cas9 gene-editing technology to enhance crops and
breed novel cultivars.
2. Development of CRISPR/Cas9 technology
In 1987, CRISPR was identified by Nakata research group at Osaka
University in E. coli and reported five 29-nucleotide repetitive palindromes in
the 30-flanking sequence of the iap gene in E. coli [4]. CRISPR was reported
as an immune defence mechanism to fight against viral and plasmid DNA.
The genes flanking CRISPR loci are found to be associated with bacterial
immune system. The CRISPR loci has three elements: at 5’ end a trans-
activating RNA (tracrRNA) gene, a Cas protein encoding gene in the middle
and a CRISPR locus at the 3’ end, which is mainly composed of a lead
sequence followed by a 23-50 bp repeat sequence and a spacer sequence. The
interval sequence consists of 17-84 bp, with an average length of
approximately 36 bp (Figure 1) [5].

Fig 1: Basic structure of CRISPR/Cas9 (Downloaded from Liu et al. 2022).

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 The CRISPR/Cas system is further divided into different categories based
on the number of Cas proteins and their structure and function. On the basis
of number of Cas proteins there are two categories i.e. Class I and Class II and
according to the structure and function of Cas protein it is subdivided into six
types (Type I to Type II). Class I includes Type I, Type III, and Type IV,
whereas Class II includes Type II and Type V. In Class II systems only one
Cas protein is required and that is why most commonly used CRISPR/Cas
gene editing systems belongs to Class II alongwith those involving Cas9, Cpf1
(Cas12a) without a tracrRNA and Cas13 with RNA cutting activity [6].
Additionally, Cas9 contains the HNH domain and the RucV-like domain. The
HNH domain cuts complementary strands of CRISPR RNA (crRNA),
whereas the RucV-like domain cuts non-complementary strands, resulting in
DNA double-strand breaks (DSBs). Unlike the Cas9 protein, Cpf1, which is
the DNA endonuclease in a novel CRISPR-based system, contains a RuvC
domain and lacks the HNH domain [7]. Moreover, the editing of the genome
by Cpf1, which uses a guide RNA (gRNA) that is significantly different from
that used by Cas9, may be regulated at the post-translational level. The fact
that it specifically recognizes and cuts DNA to produce DSBs makes the
CRISPR/Cas9 system appropriate for editing target genes.
The mechanism of action of CRISPR/Cas9 is pretty clear after more than
a decade of research. TracrRNA and crRNA interact to produce a complex
that binds and guides Cas9 to cut the DNA sequence at a specific genomic
region in the CRISPR/Cas9 system. The researchers preceded one step further
and turned the tracrRNA and crRNA complex into sgRNA, which has only
one strand of RNA. Cas protein identifies a conserved sequence while being
guided by sgRNA [8]. It identifies and binds to the protospacer adjacent motif
(PAM) and unwinds double-stranded DNA. The crRNA is a complementary
sequence that binds to the target sequence located upstream of the PAM. Both
crRNA and tracrRNA derived from the CRISPR locus and forms a sgRNA
through base complementation and pairing. The sgRNA further pairs with the
sequence upstream of the PAM, resulting in a double strand break. The breaks
can be repaired by non-homologous end joining (NHEJ) or homology-directed
repair (HDR). The NHEJ repair mechanism sometimes leads to gene
mutations as it is more prone to errors. HDR precisely inserts or replaces bases
to repair the break site in presence of donor DNA (Figure 2) [9].

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Fig 2: Genome editing at the CRISPR/Cas9 target locus (Downloaded from Liu et al.
2022).
3. Gene editing tools based on CRISPR/Cas9
CRISPR/Cas9 has emerged as one of the most adaptable technologies for
editing particular genes in various genomes in recent years. Many
CRISPR/Cas9-based tools have been developed, allowing researchers to alter
genes in a variety of ways (for example, gene knockout, gene knock-in, gene
regulation, base editing, and prime editing).
3.1 Gene knockout
Gene knockout approaches are critical for evaluating and confirming gene
functioning and the changes in biological traits that results. CRISPR/Cas9
gene knockout vectors, for example, are generally developed and introduced
into plants. Target-gene knockout mutants can be obtained after one or two
generations of screening [10]. A single target gene can be knocked out, or
multiple target genes can be knocked out at the same time. Using Golden Gate,
Gibson, or isotail polymer methods, several sgRNAs can be integrated into a
binary vector [11]. The efficiency of gene editing can be increased by targeting
several regions in a single gene of interest.

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3.2 Gene knock-in
After cleaving the DNA with Cas, gene knock-in techniques are
employed to insert a foreign DNA fragment into a particular genomic location.
Two major DNA repair processes (HDR and NHEJ) are used following the
insertion of a DSB into the genome, depending on the traits of the donor vector
and the cell cycle stage. The cellular DNA break repair pathways may increase
the usefulness and effectiveness of gene insertion techniques, as CRISPR-
based gene knock-in methods primarily involve either homology-independent
strategies (NHEJ-based targeted insertion events) or homology-dependent
strategies (HDR-based targeted insertion events) [12].
3.3 Base editing
Single-base editors and double-base editors are both types of base editors.
The first base editors were the single-base editors i.e., cytosine base editors
(CBEs) and adenine base editors (ABEs), which both mediate only one sort of
base transition (i.e. C-to-T and A-to-G, respectively). Consequently, their
applicability for site saturation mutagenesis is restricted [13]. CBEs and ABEs
were useful in making CRISPR/Cas-based genome-editing technologies more
broadly available [38]. These base editors are capable of changing genomic
bases without causing DSBs and requiring NHEJ/HDR. Editors based on
human APOBEC3a can efficiently change C to T in wheat, rice, and potato
[14]
.
3.4 Gene regulation
Gene regulation is divided into two categories: transcriptional regulation
and post-transcriptional regulation. Transcriptional regulation encompasses
genetic DNA regulation and epigenetic chromatin regulation [15]. Death Cas9
(dCas) was obtained by modifying the Cas protein which lacks nuclease
activity but can recognize specific DNA sequences. Similar to how
transcription factors bind to target gene promoters, dCas binds to double-
stranded target genes. Despite this, dCas cannot work by itself but can
sterically hinder other molecules. To modify gene transcription, researchers
have paired dCas with transcriptional activators/suppressors [16]. When dCas
attaches to the target gene's promoter or transcription start point, it can inhibit
transcription from initiating. When dCas attaches to the target gene open
reading frame, it blocks RNA polymerases and transcription factors from
binding and thus inhibits transcription. Furthermore, CRISPR/Cas9 regulates
gene expression by epigenetic control involving chromatin, gene
modifications following dCas binding to DNA methylase and acetylase, or by
altering chromatin structure and modulating the interaction between the
enhancer and promoter to regulate gene expression.

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4. Applications of CRISPR/Cas9 in crop improvement
The primary purpose of genetics and plant breeding research is to
determine the relationship between genotype and phenotype. Traditional plant
breeding relies heavily on phenotype and the breeder's experience to pick
improved types. This is a major challenge to crop improvement. The
advancement of high-throughput sequencing technology has increased the
availability of sequenced crop genomes, which has considerably aided in the
study of gene functions and the mining of genes that regulate important traits
such as yield, quality, stress tolerance, and disease resistance. Gene function-
related research has identified key regulatory genes and the networks that
control complex crop traits. In addition, some of these genes have been
precisely modified in order to improve germplasm resources and progressively
build accurate molecular breeding systems. In this context, CRISPR/Cas9 can
be used to increase a wide range of crop attributes, including yield, quality,
stress tolerance, disease resistance, and herbicide resistance, in order to
generate a large amount of new germplasm.
4.1 Improvement of crop yield
Yield improvement is one of the major traits considered during crop
improvement. On the basis of CRISPR/Cas9 technology, recent research has
discovered genetic modifications that have enhanced the output of rice, wheat,
and other food crops. Miao et al. (2018) generated a triple knockout rice
mutant pyl1/4/6 using CRISPR/Cas9 system. The mutant had higher yield,
longer panicles, more primary and secondary branches and fewer tillers per
plant as compared to wild-type control [17]. In another research, the rice grain
weight increased considerably after three grain weight-associated genes were
concurrently deleted (GW2, GW5, and TGW6) [18]. An amino acid transporter,
OsAAP3 associated with the allocation of nutrients in rice has been knocked
out, which significantly increases the number of tillers and grain yield without
disturbing the grain quality [19]. Knocking out another transporter OsSNB,
which is responsible for regulation of floral development, can increase grain
length and width along with 1000-grain weight. Zhang et al. (2016) observed
that the 1000-grain weight increased in the T0 mutant by targeting TaGASR7
by transiently expressing CRISPR/Cas9 DNA or RNA in the calli of hexaploid
wheat and tetraploid durum wheat [20].
Knocking out TaGW2 gene encoding a RING E3 ligase in wheat
increases the grain yield by increasing the grain length and width. The grain
yield in wheat increases after deleting TaCKX2-D1, which encodes cytokinin
oxidase/dehydrogenase in wheat [21]. The CRISPR/Cas9 mediated knockout of

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the terminal of OaLOGL5 coding region increase the number of grains, grain
weight per spike and total yield in rice [22].
4.2 Improvement in crop quality
The demand for high-quality crops is rising as living standards around the
globe generally rise. The quality of a crop, which is determined by both
exterior and interior features, has a significant impact on its market value.
Physical features like as size, colour, texture, and aroma are crucial
determinants of crop quality. Specific nutrients (such as proteins,
carbohydrates, and fats) and bioactive compounds (such as carotenoids,
lycopene, aminobutyric acid and flavonoids) have an impact on crop traits that
are connected to internal quality [23].
Crop quality has significantly increased as a result of recent research
using the gene-editing CRISPR/Cas9 system. Because of their nutritional and
culinary benefits, grains with low amylose concentrations are recommended.
They are also extensively employed in the adhesive and textile sectors. Rice
grains can have their amylose content reduced while also having their
nutritional value and flavour enhanced using the CRISPR/Cas9 technology.
Ma et al. obtained waxy rice mutants by knocking out the WAXY (Wx) gene
and successfully reduced the amylose content from 14.6% to 2.6% [11]. Maize
gene Wx1 encodes starch synthase and knocking out this gene via
CRISPR/Cas9 system can increase the amylopectin content of grains to almost
100%. Sun et al. (2017) knocked out the SBEIIB gene, which increases the
starch content from 1% to 9.8% [24]. Aroma is an important quality of rice
grains. Knocking out the gene encoding betaine aldehyde dehydrogenase
(OsBADH2) using TALENs and the CRISPR/Cas9 system can enhance the
flavor of rice grains [25].
The nutritional composition of crops has been improved by the
CRISPR/Cas9 system, which has increased the yield of high-oleic-acid
soybean, the starch quality of potato and gluten-free wheat, the lycopene and
aminobutyric acid content of tomato, the carotenoid content of rice, and the
oil content of soybean [26]. Dahan et al. (2018) edited the CRTISO gene in
tomato, which significantly increased the carotene content [21]. Cermak et al.
(2015) inserted the Cauliflower mosaic virus 35S promoter into the promoter
region of the tomato ANT1 gene via a homologous recombination involving
the twin virus replicator to specifically activate ANT1 expression and increase
the fruit anthocyanin content by several fold. Additionally, tomato fruits with
increased post-harvest longevity have been developed using CRISPR/Cas9
technology [27]. The examples of genes from various crops that have been

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modified by the CRISPR/Cas9 system to increase crop yield and quality are
shown in Table 1.
Table 1: Improvement in crop yield and crop quality using different CRISPR/Cas9
methods. (Reproduced from Liu et al., 2022).

Editing
Crop Gene Gene function Features
method
PYL1, PYL4, Regulated plant Promote rice growth and
Rice Knockout
PYL6 growth productivity
Negative regulators
GW2, GW5 controlling yield-
Rice Knockout Increased grain weight
and TGW6 associated
characteristics of rice
An amino acid osmotic
Higher tiller number and
Rice OsAAP3 enzyme related to Knockout
grain yield
nutrient allocation
Grain length and
Wheat TaGASR7 Knockout 1000-grain weight
weight
Encoding RING E3 Increased the length and
Wheat TaGW2 Knockout
ligase width of wheat grains
Encoding cytokinin Increased grain number
Wheat TaCKX2-D1 Knockout
oxidase/dehydrogenase and wheat yield
Increased grain number
Cytokinin activating
Rice OsLOGL5 Knockout and weight per spike as
enzyme
well as the yield of rice
Increased maize
Encoding starch
Maize Wx1 Knockout amylopectin content close
synthase
to 100%
Determined the fine
SBEI, Increased AC and RS
Rice structure and physical Knockout
SBEIIb content
properties of starch
Spring Catalyzed synthesis of
Waxy Knockout Reduced amylose content
barley amylose
in-situ
Regulated plant site- Increased anthocyanin
Tomato ANT1
growth specific content
activation
Reduced the contents of
Encoding FATB
Soybean GmFATB1 Knockout two saturated fatty acids
protein
in soybean

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4.3 Improvement in crop disease resistance
There has been a significant amount of research done recently on using
the CRISPR/Cas9 system to make crops more resistant to fungi, bacteria, and
viruses. To improve plant disease resistance, the following techniques are
typically employed to edit the genome and change particular plant defence
mechanisms.
Plant resistance to fungal infections has been increased using
CRISPR/Cas-based editing technologies. Increased resistance to powdery
mildew is a result of a loss-of-function mutation in MILDEW LOCUS O
(MLO), which was first discovered in barley and later recognised in
monocotyledons and dicotyledons as a typical S gene. Mutations in MLO gene
can also leads to disease resistance in wheat and tomato. In wheat, there was
enhanced resistance to powdery mildew by inducing mutation in TaMLO-A1
using CRISPR/Cas9 [28]. In tomato, sixteen SlMLO alleles have been found,
with SlMLO1 being the most crucial for disease resistance. Tomelo, a new
non-transgenic tomato variety highly resistant to powdery mildew was
developed by selfing. Knocking out OsERF922 and OsSEC3A using
CRISPR/Cas9, generated rice blast resistant mutants [29].
Bacterial diseases, in addition to fungal diseases, have a significant
impact on agricultural output and quality. In order to infect rice plants,
Xanthomonas oryzae pv. oryzae (Xoo) stimulates the production of a sucrose
transporter gene. To create rice lines with broad-spectrum resistance to several
Xoo physiological races, the promoter regions of Sweet11, Sweet13, and
Sweet14 were simultaneously edited using the CRISPR/Cas9 genome-editing
technique. By altering the PthA4 effector-binding region in the CsLOB1
promoter, a mutant resistant to the citrus ulcerative pathogen Xanthomonas
citri subsp. citri (Xcc) was created in grapefruit [30]. In order to boost the
resistance to fire blight, Malnoy et al. (2016) edited the apple protoplasts'
DIPM-1, DIPM-2, and DIPM-4 genes using CRISPR/Cas9 ribonucleoproteins
(RNPs) [31]. These researches suggest that crop resistance to bacterial
infections can be increased using CRISPR/Cas-based editing technologies.
The yield and quality of crops can also be dramatically reduced by viral
infections. CRISPR/Cas9 has thus been employed to increase resistance
against viral infections. CRISPR/Cas9 system involving Cas9 or Cas13a has
been used to impart resistance to DNA or RNA viruses. 11 sgRNA were
designed which target the sequence encoding the replication initiation protein
motif, replication binding site, hairpin structure and non-nucleotide sequence
to reduce the viral load of soybean yellow dwarf virus in tobacco by 87% [32].

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To dramatically limit the reproduction and accumulation of the virus, Ali et
al. created sgRNAs that specifically target conserved stem-loop regions
unique to the coding and non-coding sequences of Tomato Yellow Leaf Curly
Virus [33]. Francisella novicida Cas9 (FnCas9) and Cas13a are two Cas9
variants that have the potential to directly target and destroy RNA.
Additionally, tobacco and A. thaliana were used to express Cucumber mosaic
virus (TMV) RNA-targeting sgRNA and FnCas9 vectors. In comparison to
the control, the accumulation of CMV and TMV in the resulting transgenic
lines decreased by 40% to 80%. Moreover, the sgRNA-FnCas9 system's
attained resistance was stably inherited [34].
4.4 Improvement in abiotic stress tolerance
Abiotic stressors, such drought, salt, extreme heat, and soil pollution,
have a significant negative impact on crop growth and significantly impede
attempts to improve agricultural output and quality. Research using
CRISPR/Cas9-mediated gene editing to increase crops' ability to withstand
abiotic stress is moving along quickly. A mutation in gene ZmSRL5, which is
associated with formation of cuticle in maize, can enhance drought tolerance
in maize [35]. Additionally, drought tolerance can be improved by editing the
promoter region of ZmARGOS8, which acts as a negative regulator of the
maize response to ethylene. Zhou et al. (2016) enhanced the elite rice restorer
line 'Hua Zhan's' resistance to drought, high temperatures, and osmotic stress
by reducing the line's sensitivity to abscisic acid and minimising the rate of
leaf water loss [18]. By using CRISPR/Cas9, Kumar et al. (2020) altered the
Indica rice gene related to drought tolerance; the mutants were highly tolerant
of salinity stress and moderately tolerant of osmotic stress at the seedling
stage, suggesting their approach was suitable for enhancing the drought and
salinity tolerance of indica rice varieties [36]. Lou et al. (2017) altered the
OsSAPK2 sequence and discovered that homozygous T1 OsSAPK2 mutants
were moderately resistant to abscisic acid but extremely susceptible to drought
stress, indicating a link between OsSAPK2 and rice drought tolerance. Shen
et al. (2017) employed CRISPR/Cas9 technology to precisely modify
OsAnn3; subsequent phenotypic analysis revealed that the six mutant lines
were more vulnerable to low-temperature stress than the wild-type control [37].
5. Limitations associated with CRISPR/Cas9 technology and their
solutions
In recent years, the CRISPR/Cas9 system and related gene-editing
methods have advanced quickly. Numerous reports have described the
functional annotation of crop genes and the extremely accurate molecular

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breeding of crops. The currently available CRISPR/Cas9 editing technique
still has certain limitations, including the occurrence of off-target effects,
editing scope, and restrictions related to the plant genetic transformation
system.
5.1 Off-target effects
Cas9 cleavage of non-target genomic regions results in off-target effects.
Such effects are influenced by two factors. First, non-target sequences may be
targeted by sgRNA. For CRISPR/Cas9 techniques to be effective, sgRNA
must bind to its target sequence with specificity. However, due to the
complexity of genomes, sgRNA may also attach to related regions in addition
to the target location. As a result, off-target effects will emerge from the
localised activation of the Cas9 endonuclease, which will result in the
cleavage of non-target sites. Second, Cas9 might pick up on a non-standard
PAM. Cas9 should cut three bases upstream of the PAM site in the
CRISPR/Cas9 system. However, occasionally Cas9 detects both a non-
standard PAM and a conventional PAM close to the target site, leading to off-
target effects. In order to reduce the likelihood of off-target events, non-
standard PAMs must be considered while creating the target sequence.
Increasing Cas9's specificity and fidelity may help to reduce the off-target
consequences associated with Cas9. Studies on the off-target effects of
CRISPR/Cas9 using GUIDE-Seq and Digenome-Seq have demonstrated that
off-target locations can be predicted by analysing the genome-wide DSBs
brought on by Cas9 [38]. Identifying DSBs in the genome caused by the
CRISPR/Cas9 system, on the other hand, remains difficult. By altering
specific enzyme regions, some researchers have been able to limit SpCas9's
off-target effects. To obtain SpCas9-HF, Asn 497, Arg 661, Gln 695, and Gln
926 in SpCas9 were transformed to Ala, and then GUIDE-Seq was utilised to
evaluate off-target sites throughout the genome. The results showed that
SpCas9-HF had considerably fewer off-target events than SpCas9 [39]. Cas9 is
made up of several domains that each has a different purpose. To create Hy-
paCas9 with improved specificity, we altered the SpCas9 REC3 domain,
which identifies complementary strands generated by sgRNA and target
sequences, controls the HNH nuclease, and regulates overall catalytic activity.
Cas9 specificity can be increased by optimising its structure. Furthermore,
decreasing the in vivo Cas9 activation period can help to prevent off-target
consequences. For example, the RNP combination of sgRNA and Cas9 can be
supplied directly into the cell, or Cas9 can be broken into two halves and then
forced to rejoin into the intact Cas9 protein within the cell by small molecules
[40]
.

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5.2 Target site limitations
The CRISPR/Cas9 technology has sped agricultural trait development;
however, the target sites of the CRISPR/Cas9 nuclease are constrained by the
associated PAM sequences, making editing all target loci challenging. As a
result, correctly changing Cas9, boosting the compatibility of the CRISPR
nuclease with various PAMs, and expanding the genomic region editable by
CRISPR/Cas9 will determine whether the CRISPR system can be widely used
to modify crops in the future. Target range of Cas9 can be increased by
extending the PAM recognition range. Different variants of Cas9 have
emerged that recognize different PAM sequences which leads to wider
genomic range. Keith’s laboratory in 2015 developed a mutant SpCas9-VRQR
and SpCas9-VRER that recognizes different sequences [41]. As a result of
technological advancements, all PAMs are now recognised by SpCas9 and its
variants, broadening the range of crop gene editing.
5.3 Foreign genes
Plant transformation with CRISPR/Cas9 gene-editing vectors by
Agrobacterium may result in the random integration of vector fragments into
plant genomes, resulting in the introduction of foreign genes and possible
biosafety problems [42]. Lines missing external genetic material in crops that
reproduce sexually can be screened via genetic separation of sexual
generations. However, for crops that reproduce asexually, genetic separation
makes the isolation and removal of foreign genes impossible. It is possible to
use the RNP complex to crops without the necessity for genetic separation of
sexual generations if it is employed to avoid importing foreign genes. This
procedure, however, is relatively complex to complete, necessitating more
technical refinements to increase its efficiency and usability for additional
crops. Transgene-free plants can be generated using RNP transfection,
transient transgene expression, and nanobiotechnology-based techniques. In
addition, an efficient tool for evaluating the biological safety of genome-edited
products has been established. There is an online tool for detecting foreign
elements in genome-edited organisms. It can be utilised when no information
about the foreign carrier component is available. This tool searches whole-
genome sequencing data for 46,695 exogenous components [43].
6. Summary
Domestication of new crops is a rather slow process. However,
throughout the domestication stage, over 7000 crops were grown throughout
the world, which created the groundwork for the production of current
cultivars. Cross breeding came into existence in the late 19th century. These

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breeding techniques i.e., cross breeding, heterosis breeding and mutagenesis,
still rely on breeders choosing materials based on phenotypic analyses, but
they have significantly enhanced crop yields and reduced the food deficit
brought on by the significant growth in the world's population over the
previous century. Additionally, the usefulness of traditional breeding methods
may be somewhat constrained for complex traits. Therefore, developing new
varieties that are stress resistant and have good yields and quality is a difficult
task. Molecular breeders have created and used gene-editing technologies to
supplement conventional breeding methods, maintain sustainable agricultural
production, and create new crops that effectively utilise environmental
resources (such as nutrients and water) and resist biotic and abiotic challenges.
Therefore, rather than replacing conventional breeding techniques, the
development of gene-editing technologies should complement them. Gene-
editing technology's cost and effectiveness mostly depend on two aspects.
Prior to anything else, particular methods must be devised and optimised, such
as transformation systems. The use of gene editing technology will expand
with the development of low-cost, low-risk, and effective transformation
systems according to various crop features. The second aspect is the regulatory
actions of the government. More precisely, there is a global dispute on
whether CRISPR-edited varieties should be subject to the same laws as
conventional genetically modified crops or be permitted to be sold
unrestricted. When commercialising new CRISPR-modified plants, it is
crucial to consider the cost of the regulation, which will be impacted by the
quantity of safety-related data needed [44].
Along with the effects of climate change and population growth,
urbanisation reduces the amount of arable land that is accessible for food
production, which presents numerous challenges. To create novel traits and
varieties that can boost yields or withstand harsh environments, cutting-edge
plant breeding techniques, such as CRISPR/Cas9, are required. Using the
CRISPR/Cas9 technology, researchers have successfully altered and enhanced
numerous quality-related features. Additionally, several gene-edited crops,
such as TALEN-fad2 soybean, TALEN-ppo potato, and CRISPR-wx1 maize,
have been commercialised, indicating that the underlying technology has
moved past the proof-of-concept research stage [94]. Genome sequencing data
for a growing variety of crops are now accessible owing to ongoing
advancements in sequencing technology and reducing sequencing costs,
which lays the groundwork for future crop gene editing. Increases in
agricultural productivity, quality, disease resistance, and many other
phenotypic traits have been attributed to CRISPR/Cas9 technology, which
allows for the quick and accurate editing of genes in a variety of crops. In

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addition, its implementation has reduced the need for water use, labour, and
the use of pesticides and fertilisers. As a result, this technology will support
the growth of sustainable agriculture and be crucial in resolving future crises
associated with food scarcity.
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Int. J. Mol. Sci. 2021; 22: 11753.

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 Chapter - 9
Various Methods of Testing Hypotheses and
Measure of Dispersion in Statistics

Author
Shiv Kumar Rana
Ph.D. Scholar, Department of Agricultural Statistics

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Page | 144
 Chapter - 9
Various Methods of Testing Hypotheses and Measure of
Dispersion in Statistics
Shiv Kumar Rana

Testing a hypothesis or presumption in respect to a statistical parameter is

known as hypothesis testing. To determine if a hypothesis is acceptable or not,
analysts use hypothesis testing.
Hypothesis testing is a crucial stage in statistics since it requires the
confirmation of an assumption that may be used to create a statistical
parameter. As an illustration, a researcher may develop a hypothesis based on
the assumption that the average of all odd integers is an even number.
The researcher will need to evaluate the hypothesis using hypothesis
testing techniques in order to determine whether it is acceptable. Hypothesis
testing is necessary to have credible evidence in order to prove that a statistical
hypothesis is true, unlike a hypothesis that is "said" to hold true on the basis
of little or no evidence.
Maybe this is where statistics come into play. This method involves a lot
of elements. However, we must first comprehend the many kinds of
hypotheses that are engaged in the process before we can comprehend the
steps involved in hypothesis testing in research technique.
Methods of hypothesis testing
1. Frequentist hypothesis testing
The frequentist hypothesis, sometimes referred to as the conventional
approach to hypothesis testing, is a technique for testing hypotheses that
focuses on forming assumptions while taking recent evidence into account.
Based on the available information and a pair of hypotheses, the alleged
truths and assumptions are made. Null Hypothesis Significance Testing is a
widely common variant of the frequentist method (NHST).
Since its introduction in the mid-1950s, the NHST technique (which
includes the null and alternative hypothesis) has been one of the most popular
approaches for testing a hypothesis in the field of statistics.

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 The following components are present in the most widely used frequentist
hypothesis tests:
1) Assumptions about the model (e.g., for the t-test for the mean, the
model assumptions can be phrased as: simple random sample of a
random variable with a normal distribution).
2) Null and alternative hypothesis.
3) A study statistic. Extreme test statistic results must cause doubts
about the validity of the null hypothesis for this to be true.
4) A mathematical theory stating that “the test statistic's sampling
distribution has the following shape if the null hypothesis and model
assumptions are both true.”
Depending on the individual hypothesis test, the specifics of these four
components may vary. We'll use an example to explain.
Example
The elements of the large-sample z-test for the mean are as follows:
1. Model assumptions: We are working with straightforward random
samples of the normal-distributed random variable X.
2. Null hypothesis: A particular number, 0 is the mean of the relevant
random variable. The alternative hypothesis may be "The mean of
the random variable X is not 𝜇0 ." "The mean of the random variable
X is less than 𝜇0 " or "The mean of the random variable X is bigger
than 𝜇0 ." We'll take the first possibility in this instance, "The mean
of the random variable is not 𝜇0 " to explain the situation. (This is the
two-sided alternative).
3. Test statistic: 𝑥̅
We now take a step back and think about all conceivable size n
simple random samples of x. The value of the x-bar is obtained for
each n-by-n simple random sample of x. 𝑥̅ is our new random
variable as a result. (𝑥̅ denotes the new random variable.) The
distribution of 𝑥̅ is called the sampling distribution of 𝑥̅ .
4. According to the theory, if all model assumptions are accurate and x
has a mean of 𝜇0 , the sampling distribution is normal, with a mean of
𝜎
𝜇0 and a standard deviation of , where 𝜎 (sigma) is X's random
(√𝑛)
variable's standard deviation. (Note: 𝜎 is called the population
standard deviation of X; it is not the same as the sample standard
deviation s, although s is an estimate of 𝜎.)

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2. Bayesian hypothesis testing
The Bayesian Hypothesis Testing, a far more novel and contemporary
approach to hypothesis testing, asserts that it will evaluate a given hypothesis
in light of both recent data that support it and historical data samples, or prior
probability.
The outcome gained shows how likely the theory is after all. In this
approach, hypothesis testing is done immediately and is based on "prior
probability and posterior probability."
The Bayesian technique evaluates a hypothesis to determine whether it is
true or incorrect based on this prior probability. A key element of this
methodology is the Bayes factor, which shows the probability ratio between
the null hypothesis and the alternative hypothesis.
Dispersion
Averages (or measures of central tendency) indicate the concentration of
observations near the centre of the distribution. As the following example
shows, knowing only the average does not give us a complete view of the
distribution.
Consider the series (i) 1,5,9,13,17, (ii) 3,6,9,12,15, and (iii) 7,8,9,10,11.
In each of these cases, the number of observations (n) is 5 and the mean is 9.
We cannot determine if the average of the first series, second series, third
series, or any other series of five observations whose sum is 45, if we are given
that the mean of five observations is 9. As a result, it is clear that the
measurements of central tendency are insufficient to fully describe the
distribution. Other actions need to be taken in order to complement and
enhance them. Dispersion is one such measure.
Dispersion means 'scatteredness' in the literal sense. We study dispersion
to understand the nature of the distribution's homogeneity or heterogeneity.
The following are some key definitions of dispersion:
i) “Dispersion of the measure of extent to which individual items vary.”
- L.R. Connor
ii) “The measure of the scatteredness of the mass of figures in a series
about an average is called the measure of variation or dispersion.”
- Simpson and Kafka
iii) “The degree to which numerical data tend to spread about an average
value is called variation or dispersion of the data”
- Spiegel

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Measures of dispersion
The scattering of data is indicated by a measure of dispersion. It explains
the discrepancy of data from one another, providing a precise understanding
of their distribution. The measure of dispersion illustrates and informs us
about the fluctuation and central value of a single object.
In other word, dispersion is the degree to which values within a
distribution deviate from the distribution's mean. It provides us an indication
of how much different objects differ from one another and from the central
value.
1. Types of measures of dispersion
1) Absolute measures
• Absolute dispersion measurements are represented in the variable's
own unit, such as kilogram’s, rupees, centimeters, or marks.
2) Relative measures
• Relative measurements of dispersion are calculated as ratios or
percentages of the average.
• These are also known as coefficients of dispersion.
• These are just numerical values or percentages that don't depend at
all on the units.
2. Absolute measures of dispersion
Following are the different absolute measures of dispersion:
a) Range
• It is the simplest way of measuring dispersion.
• It is defined as the difference between the largest and smallest
item in a particular distribution.
Range = Largest item (L) - Smallest item (S)
b) Inter-quartile range
• It is defined as the difference between the upper and lower
quartiles of a particular distribution.
Inter-quartile range = Upper Quartile (Q3) - Lower Quartile (Q1)
c) Quartile deviation
• It is referred to as the Semi-Inter-Quartile Range, which is half
the difference between the upper and lower quartiles.

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 Upper quartile (Q3 ) − Lower quartile (Q1 )
Quartile deviation =
2
d) Mean deviation
• The term "mean deviation" refers to the arithmetic average
(mean) of observations' deviation from a mean or median value.
e) Standard deviation
• The square root of the arithmetic mean of the squared deviation
from the mean is the standard deviation.
f) Lorenz curve
• This is a graphic method of measuring estimated dispersion.
• This curve is frequently used to estimate the wealth or income
disparities in a community.
3. Relative measures of dispersion
The relative measures of dispersion are as follows:
a) Coefficient of range
• It is the ratio of the difference between two extreme items in a
distribution to the total of their sums.
(Largest item − Smallest item)
Coefficient of range =
(Largest item + Smallest item)
b) Coefficient of quartile deviation
• It describes the relationship between the difference between an
upper and lower quadrant of a distribution and their total.
Q 3 − Q1
Coefficient of quartile deviation =
Q 3 + Q1
c) Coefficient of mean deviation
• Mean deviation is a measure of absolute dispersion.
• To convert it to a relative measure, divide it by the average from
which it was computed. It is then known as the Coefficient of
Mean Deviation.
• Coefficient of Mean Deviation from Mean
𝑀𝐷𝑋̂
(𝑋̂) =
𝑋̂

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 • Coefficient of Mean Deviation from Median
𝑀𝐷𝑀𝑒
(𝑀𝐸) =
𝑀𝑒
d) Coefficient of standard deviation
• It is computed by dividing the standard deviation (σ) by the
mean (X ̅) of the observation.
• Coefficient of standard deviation
σ
S. D. =
̅
X
e) Coefficient of variation
• It is employed to assess the stability of two sets of data (or
uniformity or consistency or homogeneity).
• It expresses the percentage relationship between the standard
deviation and the arithmetic mean.
σ
𝐶. 𝑉. = × 100
𝑋̅
• Where, C.V. = Coefficient of variation, σ=
̅
Standard deviation, X = Arithmatic mean
Characteristics of measures of dispersion
The criteria for an ideal measure of dispersion are the same as those for
an ideal measure of central tendency, namely,
i) It should be precisely specified.
ii) It should be simple to calculate and understand.
iii) It must be founded on all observations.
iv) It should be mathematically solvable.
v) It should be as unaffected as feasible by sampling variations.
Merits and demerits of range
Merits
1) It is extremely simple to compute and understand.
2) No particular knowledge is required to calculate range.
3) It is the quickest to compute.
4) It gives a high-level overview of the facts at a glance.

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Demerits
1) Since it is primarily based on two extreme numbers, it is an imperfect
measure (highest and lowest).
2) It cannot be computed when the series is open-ended.
3) Range varies greatly from sample to sample and is heavily impacted
by sampling changes.
Merits and demerits of quartile deviation
Merits
1) It is also quite simple to compute and understanding.
2) It may be applied to open-end distribution as well.
3) It is preferable than "Range" since it is less impacted by extreme
numbers.
4) It is more helpful when computing the dispersion of the middle 50%.
Demerits
1) It does not consider all of the observations.
2) It cannot be further algebraically or statistically treated.
3) It is significantly influenced by sampling variations.
4) It is not considered as a particularly trustworthy measure of
dispersion since it neglects 50% of the data.
Merits and demerits of mean deviation
Merits
1) It is based on all of the series' observations rather than just the
limitations such as Range and QD.
2) It is straightforward to compute and understand.
3) It isn't significantly impacted by extreme values.
4) Deviations from any average can be used to determine mean
deviation.
Demerits
1) From a mathematical perspective, it is undesirable to disregard + and
- signs. It is not capable of further mathematical treatment.
2) When the mean or median is expressed as a fraction, computation is
complicated.
3) In the event of open-ended series, it might not be possible to apply
this method.

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Objectives of computing dispersion
1) Comparative study
• A single value produced by a measure of dispersion indicates the
level of consistency or homogeneity of a distribution. We may
contrast several distributions with just single value.
• The amount (value) of dispersion decreases as consistency or
uniformity increases, and vice versa.
2) Reliability of an average
• A lower value of dispersion indicates that there is less variation
between the observations and the average. It suggests that the average
is a reliable and useful method of representing observation.
• A larger dispersion number indicates a greater degree of deviation in
the data. The average is not a useful measure and cannot be relied
upon in this case.
3) Control the variability
• Different dispersion measures provide us information on variability
from numerous perspectives, and this knowledge may be helpful in
controlling the variance.
• These measures of dispersion might be very useful, especially when
conducting financial analyses in the fields of medicine and business.
4) Basis for further statistical analysis
• Measures of dispersion provide the basis for further statistical
analysis, including the computation of correlation, the estimation of
regression, the testing of hypotheses, etc.
Conclusion
The Frequentist technique or the Bayesian approach can both be used for
hypothesis testing, which is a means to determine whether a presumption is
plausible. The frequentist method makes assumptions without considering a
probability, whereas the Bayesian approach does. Test statistic, significance
level, p-level, and hypothesis testing technique are some of the components of
hypothesis testing. Verifying the data samples and choosing the most likely
hypothesis from the null hypothesis and alternative hypothesis is an important
approach to evaluate if a hypothesis is true or not.
Dispersion in statistics refers to the measure of the variability of data or
terms. Such variability may give random measurement errors where some of

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the instrumental measurements are found to be imprecise. It is a statistical way
of describing how the terms are spread out in different data sets.
References
a) Books
1. Gupta, S.C. and Kapoor, V.K. 2014. Fundmentals of Mathematical
Statistics. 4th edition. New Delhi, Sultan Chand & Sons. 2.12-2.14.
2. Rangaswamy, R. 2010. A Textbook of Agricultural Statistics. 2nd
edition. New Delhi, New Age International Publishers. 41-48.
3. Agarwal, B.L. 2022. Programmed Statistics. 4th edition. New Delhi, New
Age International Publishers. 60-64.
b) Website
1. [Link]
2. [Link]

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 Chapter - 10
Emerging Challenges of INM in Crop
Management

Authors
Saloni Yadav
Ph.D. Scholar, Department of Soil Science
Amit Kumar
Ph.D. Scholar, Department of Soil Science
Asha
Ph.D. Scholar, Department of Soil Science
Mohit Sharma
Ph.D. Scholar, Department of Soil Science

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 Chapter - 10
Emerging Challenges of INM in Crop Management
Saloni Yadav, Amit Kumar, Asha and Mohit Sharma

Abstract
Integrated Nutrient Management (INM) is a farming practice that
involves the use of a combination of different sources of plant nutrients, such
as organic and inorganic fertilizers, to optimize crop production while
minimizing negative environmental impacts. The goal of INM is to combine
the application of both organic and inorganic soil nutrients to boost crop yield
and protect soil productivity for future generations. INM can benefit farmers
by increasing crop productivity, improving soil fertility, reducing costs, and
protecting the environment. However, there are several challenges in
implementing INM such as limitations of small holdings, farmer economic
conditions, Monsoon variability, and biofertilizer limitations. Overall, INM is
an important practice in ensuring food security and rural development by
improving plant nutrient management and maintaining soil fertility.
Introduction
A significant challenge for food security and rural development is the
development of plant nutrient management, which aims to enhance the
quantity of plant nutrients in farming systems and, consequently, crop yield.
One significant but sometimes unnoticed element of land degradation is the
depletion of nutrient reserves in the soil. On the other hand, excessive nutrient
application or ineffective management results in a financial loss for the farmer
and can have an adverse effect on the environment, particularly if significant
amounts of nutrients are lost from the soil-plant system into the water or the
air. In order to keep soil fertility and plant nutrient delivery at an ideal level
for sustaining the intended productivity, integrated nutrient management
refers to the practice of optimizing the benefits from all potential sources of
organic, inorganic, and biological components in a coordinated manner. Now
a days there are lot of challenges are emerging in integrated nutrient
management such as limitations of small holdings, farmer economic
conditions and the viability of the proposed integrated nutrient management
alternative, monsoon variability, and biofertilizer limitations etc. It is very

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important to overcome these issues and focused on the utilization of both
organic and inorganic soil nutrients in combination to boost crop output while
protecting soil productivity for future generations.
Content
A nutrient is anything that an organism uses for its own existence,
development, and reproduction. They come in two categories, namely: Plants
require large amounts of some nutrients, known as macronutrients, whereas
they require smaller amounts of other nutrients, known as micronutrients.
While the air delivers gases like carbon dioxide and oxygen, water supplies
hydrogen. Iron, copper, zinc, chlorine, boron, manganese, calcium,
magnesium, potassium, nitrogen, and phosphorus are among the thirteen extra
macronutrients and micronutrients the soil supplies. Plants can't develop
without these vital nutrients, which also affect their ability to operate properly
and resist disease. Soil fertility can be increased by adding nutrients in the
form of manure and fertilizers. Integrated nutrient management (INM) is a
process for ensuring that the soil fertility and plant nutrient supply are both
kept at their ideal levels to support the intended crop productivity. The
objective of integrated nutrient management is to combine the application of
both organic and inorganic soil nutrients to boost crop yield and protect soil
productivity for future generations. Nutrient management planning places a
lot of emphasis on preventing overapplication of nutrients in order to preserve
water quality, reduce environmental effect, and still produce the best yield
possible for economic gain. It entails keeping track of all the nutrients you
possess, figuring out which nutrients you will require, and organizing how
much, when, and where to apply each nutrient to your cropland. This entails
establishing what nutrients are needed to meet the demands of crops by first
determining what nutrients are in the soil (soil testing) and what is already
present in a crop that is growing or has been harvested.
Scope and importance of Integrated nutrient management
Integrated Nutrient Management (INM) is a farming practice that
involves the use of a combination of different sources of plant nutrients, such
as organic and inorganic fertilizers, to optimize crop production while
minimizing negative environmental impacts.
1) Increased crop productivity: INM can help to increase crop
productivity by providing the right balance of nutrients for optimal
growth and development.
2) Improved soil fertility: INM practices, such as the use of organic
fertilizers, can help to improve the fertility of the soil and promote
sustainable agriculture.

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 3) Reduced costs: INM can help farmers to reduce costs by reducing
the amount of inorganic fertilizers required and by improving the
efficiency of fertilizer use.
4) Protection of the environment: INM can help to reduce the
negative environmental impacts of agriculture, such as soil erosion,
water pollution, and greenhouse gas emissions.
5) Better crop quality: INM can help to improve the quality of crops
by providing the right balance of nutrients and by reducing the risk
of nutrient deficiencies.
6) Sustainable agriculture: INM promotes the use of organic inputs,
thereby reducing the dependence on chemical fertilizers. This leads
to sustainable agriculture, which is essential for long-term food
security.
7) Climate resilience: INM helps in building the soil organic matter
which improves soil structure, water holding capacity, and nutrient
cycling. This helps in improving the resilience of the crops to climate
change.
8) Economic benefits: INM can lead to improved crop yields, which
can help to increase the income of farmers and contribute to rural
development.
Overall, Integrated Nutrient Management is a vital practice for
sustainable agriculture, it is essential for long-term food security, and to
improve the resilience of the crops to climate change. It is a cost-effective way
to increase crop productivity and improve soil fertility while protecting the
environment.
Components of integrated nutrient management
1) Soil analysis: A critical first step in INM is to conduct a soil analysis
to determine the current nutrient status of the soil. This helps to
identify any nutrient deficiencies and to plan for appropriate
fertilization.
2) Organic fertilizers: Organic fertilizers, such as compost, farmyard
manure, and green manures, are an essential component of INM.
They improve soil fertility and provide a range of essential plant
nutrients.
3) Inorganic fertilizers: Inorganic fertilizers, such as nitrogen,
phosphorus, and potassium, are also used in INM. They provide a

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 quick source of essential plant nutrients and can be used to
supplement organic fertilizers.
4) Biological fertilizers: INM also includes the use of beneficial
microorganisms, such as nitrogen-fixing bacteria, to improve soil
fertility and crop growth.
5) Crop rotation: INM incorporates crop rotation to improve soil
fertility and to manage pests and diseases.
6) Agronomic practices: INM also includes agronomic practices such
as proper irrigation, tillage and seed rate, to optimize the use of
nutrients and water to improve crop growth.
7) Nutrient budgeting: INM includes nutrient budgeting, which is a
systematic approach to manage the nutrient inputs and outputs in the
farming system.
8) Integrated pest management: INM also includes integrated pest
management, which is a holistic approach to managing pests,
diseases, and weeds that aims to minimize the use of chemical
pesticides.

Objectives of integrated nutrient management

1) Maintain economic yield: INM aims to optimize crop productivity
and profitability by balancing the use of fertilizers and other soil
amendments.

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2) Reduce environmental pollution: INM aims to minimize the
negative environmental impacts of agriculture by reducing the use of
synthetic fertilizers and other pollutants, and promoting the use of
sustainable and environmentally-friendly practices.
3) Integrate soil fertility restoring crops and livestock: INM aims to
improve soil fertility by incorporating crops and livestock that can
help restore soil nutrients and improve soil health.
4) Crop residue recycling: INM aims to recycle crop residues as
organic matter to improve soil fertility and reduce the need for
synthetic fertilizers.
5) Improve nutrient cycling: INM aims to improve the efficiency of
nutrient cycling in the soil, which can help reduce the need for
synthetic fertilizers and improve crop productivity.
6) Promote the use of organic manure: INM aims to promote the use
of organic manure, such as compost, as a source of nutrients for
crops, which can help reduce the need for synthetic fertilizers and
improve soil health.
7) Application of bio-fertilizers: INM aims to promote the use of bio-
fertilizers, which are natural and organic sources of nutrients that can
help improve crop productivity and reduce the need for synthetic
fertilizers.
8) Development and introduction of better-quality genotypes: INM
aims to improve crop productivity by developing and introducing
better-quality crop varieties that are more resistant to pests and
diseases and have higher yields.
9) Promote balanced use of fertilizers: Integrated nutrient
management aims to promote the balanced use of fertilizers, which
can help reduce the environmental impacts of agriculture and
improve crop productivity.
10) Prevent environment degradation: Integrated nutrient
management aims to prevent environmental degradation by reducing
the use of synthetic fertilizers and other pollutants, and promoting the
use of sustainable and environmentally-friendly practices.
11) Improve rain-water management to avoid soil and nutrient
erosion: Integrated nutrient management aims to improve rainwater
management by minimizing soil erosion and nutrient loss, which can
help improve soil productivity and reduce the need for synthetic
fertilizers.

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 12) Introduction of conservation farming practices: Integrated
nutrient management aims to improve soil productivity and reduce
the environmental impacts of agriculture by promoting conservation
farming practices such as tillage, agroforestry, and crop rotation.
13) Promote nutrient restoration: Integrated nutrient management
aims to restore soil nutrients through the use of organic matter, bio-
fertilizers, and other sustainable practices.
14) Improve soil physical properties: Integrated nutrient management
aims to improve the physical properties of soil, such as porosity,
structure and water holding capacity, which can help improve crop
productivity and reduce the need for synthetic fertilizers.
15) Enhance soil production efficiency: Integrated nutrient
management aims to improve soil productivity by increasing the
efficiency of nutrient cycling in the soil and reducing the need for
synthetic fertilizers.
16) Inspiring farmers to adapt less environment harming
techniques: INM aims to encourage farmers to adopt sustainable and
environmentally-friendly practices, which can help reduce the
environmental impacts of agriculture and improve crop productivity.

Challenges in integrated nutrient management

Integrated Nutrient Management (INM) is a holistic approach to
optimizing crop productivity and sustainability by balancing the use of

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fertilizers, organic matter, and other soil amendments. However, there are
several challenges that can arise when implementing integrated nutrient
management.
1) Macronutrient deficiency: As mentioned earlier, inadequate intake
of macronutrients such as carbohydrates, proteins, and fats can lead
to deficiency and health problems.
2) Micronutrient deficiency: Similar to macronutrient deficiencies,
inadequate intake of micronutrients such as vitamins and minerals
can also lead to health problems. In agriculture, micronutrient
deficiencies in crops can result in reduced growth and productivity.
3) Imbalance in fertilizer use: Overuse or improper use of fertilizers
can lead to imbalances in soil nutrient levels, which can result in
reduced crop productivity or even damage to the environment.
4) Gap in nutrient supply and nutrient use: There can be a mismatch
between the availability of nutrients in the soil and the amount
needed by crops, which can lead to reduced productivity.
5) Declining fertilizer response: Over time, crops may become less
responsive to fertilizers, which can lead to reduced productivity and
increased costs.
6) Declining crop productivity: This can be caused by a variety of
factors, including nutrient deficiencies, imbalances in fertilizer use,
and other environmental factors.
7) Stagnation in fertilizer production: A lack of innovation or
investment in fertilizer production can lead to stagnation in the
industry, which can make it difficult to meet the needs of farmers and
growers.
8) Change in land-man ratio: With an increasing global population,
the amount of land available for agriculture is decreasing, which can
make it more difficult to meet the food needs of a growing
population.
9) Lack of knowledge: Farmers and growers may not have the
knowledge or resources to properly implement integrated nutrient
management, which can lead to reduced productivity and increased
costs.
10) Monsoon variability: integrated nutrient management is heavily
dependent on the monsoon, which is known to be highly variable, it

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 can lead to uncertainty in the timing and amount of rainfall, which
can make it difficult to plan for and implement integrated nutrient
management effectively.
To overcome these challenges, it is important for farmers, researchers,
and policymakers to work together to develop and implement effective
integrated nutrient management strategies that consider the specific needs and
constraints of different regions and crops. This could include investing in
research and development, providing education and training for farmers, and
creating policies and incentives that support sustainable and productive
agriculture.
Limitation in integrated nutrient management
1) Lack of knowledge and awareness: Many farmers may not be
aware of INM practices, or may lack the technical knowledge
required to implement them effectively.
2) Limited availability of organic fertilizers: Organic fertilizers, such
as compost and farmyard manure, are essential components of INM,
but their availability may be limited in some regions.
3) High cost: INM practices can be more expensive than traditional
methods, as they require the use of multiple types of fertilizers.
4) Lack of government support: In some countries, government
policies and support for INM may be limited, making it difficult for
farmers to implement these practices.
5) Dependence on weather: The effectiveness of INM practices can be
influenced by weather conditions, such as rainfall and temperature.
6) Complexity in implementation: INM requires careful planning and
management, and may be complex to implement in practice.
7) Lack of precision: INM may not be able to provide accurate
information on the exact amount of nutrients required by the crop,
which may lead to over- or under-application of fertilizers.
8) Limited by soil condition: INM may not be suitable for certain soil
types, such as acidic or saline soils, which may require specific
management practices.
Integrated nutrient management practices in different types of crops
The specific practices of INM may vary depending on the type of crop
being grown.

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 For example, in cereal crops such as wheat, barley, and rice, INM may
involve the use of organic manures and composts in combination with
inorganic fertilizers to provide a balanced supply of nutrients. In addition,
cover cropping and crop rotation can also be used as part of an INM strategy
to improve soil fertility and nutrient cycling.
In horticultural crops, such as fruits and vegetables, INM may involve the
use of controlled-release fertilizers and drip irrigation to optimize nutrient
uptake and reduce nutrient loss through leaching. In addition, the use of
biofertilizers, such as rhizobia for legumes and mycorrhizae for other crops,
can also be an important part of an INM strategy in horticultural crops.
In cash crops such as coffee, cocoa, and rubber, INM may involve the use
of organic manures, green manures, and crop rotation to maintain soil fertility,
along with the use of inorganic fertilizers to provide a balanced supply of
nutrients.
Overall, the specific practices of INM will vary depending on the type of
crop being grown, as well as the specific conditions of the growing area, such
as soil type and climate. It is important for farmers to work with agricultural
experts to develop an INM plan that is tailored to their specific needs and
conditions.
Conclusion
Thus, Integrated Nutrient Management is a sustainable method of
farming. It aids in biological, water, and soil restoration in addition to
increasing agricultural productivity. Extension groups must inform farmers
about the deteriorating soil health, unsustainable output, and environmental
pollution brought on by the non-use of organics in order to promote INM in
India.
Suggested reading
1) "Integrated nutrient management: Principles and practices" by R.N.
Roy, J.K. Ladha, and R.J. Buresh. Published in Advances in
Agronomy, volume 110, 2011. This book provides an overview of
the principles and practices of integrated nutrient management,
including the use of organic and inorganic fertilizers, crop rotation,
and biofertilizers.
2) "Integrated nutrient management for sustainable crop production" by
S.K. Sharma and R.K. Gupta. Published in Journal of Soil Science
and Plant Nutrition, volume 14, 2014. This paper discusses the
importance of integrated nutrient management for sustainable crop

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 production, including the benefits of using a combination of organic
and inorganic fertilizers and the role of biofertilizers.
3) "Integrated nutrient management in rice-based cropping systems" by
M.S. Swain and J.K. Ladha. Published in Plant and Soil, volume 231,
2001. This paper reviews the current state of knowledge on integrated
nutrient management in rice-based cropping systems, including the
use of organic and inorganic fertilizers, crop rotation, and
biofertilizers.
4) "Integrated nutrient management in horticultural crops" by R.K.
Gupta and S.K. Sharma. Published in Journal of the Indian Society
of Soil Science, volume 62, 2014. This paper provides an overview
of the principles and practices of integrated nutrient management in
horticultural crops, including the use of controlled-release fertilizers
and biofertilizers.
5) "Integrated nutrient management in cash crops" by A. Khera and A.
K. Singh. Published in Agricultural Reviews, volume 37, 2016. This
paper reviews the current state of knowledge on integrated nutrient
management in cash crops such as coffee, cocoa, and rubber,
including the use of organic manures, green manures, and crop
rotation.

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 Chapter - 11
Grafting: Types, Advantages and Disadvantages
for the Crop Improvement

Authors
BC Akhilraj
Ph.D. Scholar, Department of Plantation, Spices, Medicinal
and Aromatic Crops, HC and RI, Tamil Nadu Agricultural
University, Coimbatore, Tamil Nadu, India
J Suresh
Professor and Head, Coconut Research Station, Tamil Nadu
Agricultural University, Aliyar Nagar, Pollachi, Tamil Nadu,
India

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Page | 168
 Chapter - 11
Grafting: Types, Advantages and Disadvantages for the
Crop Improvement
BC Akhilraj and J Suresh

Abstract
Modern horticulture depends on the traditional horticultural technique of
grafting. Long thought to be a strange technique and a dark secret, the practise
of fusing two live entities such that they subsequently work as one has long
been practised. This chapter explores the grafting, types, advantages and
disadvantages for the crop improvement and upcoming developments in our
knowledge of and use of contemporary grafting. Grafting continues to be a
popular and necessary way of plant cloning for a wide range of uses and a
large range of species, despite recent advancements in propagation
technology, such as the use of rooting hormones, misting of cuttings, and
micro-propagation. Grafting is being employed for herbaceous vegetables in
addition to typical tree fruit crops and woody ornamentals. Reducing the use
of chemical pesticides for disease control is a major objective of contemporary
breeding of disease-resistant rootstocks, which helps to achieve the goal of
sustainable agricultural output.
Keywords: Grafting, types of grafting, advantages, disadvantages
Introduction
Since the beginning of agriculture, several advancements in plant
propagation have been closely linked to the progressive domestication of food
crops. According to Goldschmidt (2014) grafting is the accidental or
intentional fusing of plant components so that vascular continuity is formed
between them and the resulting genetically composite or ganism performs as
a single plant. Intentional grafting is placing a previously cut shoot into an
aperture in another plant growing on its own root system. Grafting can occur
naturally or between two neighbouring complete plants or different branches
of the same plant (detached scion grafting). The branch piece or bud removed
from a donor plant that will develop into the top of the grafted plant is known
as a scion (cyon). The plant that accepts and fuses with the scion and serves

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as the root system of the grafted plant is referred to as stock, under stock, or
rootstock. The terms "stock" and "rootstock" are interchangeable, however the
latter two have different configurations inside the grafted plant. Unlike under
stock, which is used when the lower half of the grafted plant comprises both
the root system and some of the shoot system on which the scion is grafted,
rootstock denotes that only the root system of the composite plant is descended
from the original stock (Cantliffe, 2009). A grafted plant typically consists of
a singlegraft union between a stock and a single scion in the simplest scenario.
However, a three-part grafted plant is created using the double working
technique, which separates three genetically different pieces (rootstock,
interstock, and scion) into a linear series. A compound genetic system is
created by grafting two (or more) different genotypes together, each of which
retains its own genetic identity for the duration of the grafted plant's life. In
contrast to pink (hybrid) roses, a red-flowering rose grafted onto a white rose
plant will continue to produce red blooms. Contradictory reports of graft
"hybridization" have persisted, nevertheless, and fresh data on gene silencing
caused by RNA transfer across the graft union raises the possibility that
grafting may have genetic repercussions. The limitations of compatibility are
another crucial genetic factor associated to grafting. Through the history of
grafting, it has been frequently mistaken that taxomornic affinity determines
which species may be effectively grafted onto any other. In general,
intraspecific/interclonal grafts are almost always compatible,
interspecific/intrageneric grafts are frequently compatible,
intrageneric/intrafamilial grafts are seldom compatible, and interfamihal
grafts are almost never compatible. The discovery that the level of taxonomic
affinity required for compatibility varies greatly across various species
complicates these generalisations (Mudge et al., 2009).
Compatibility
Theoretically, any two botanically related plants with a continuous
cambium can be grafted together. Grafts between species are frequently
successful, those between genera are rarely successful, but those between
families are almost never successful. Although the proximity of botanical
relationships within a genus is not a foolproof predictor of success, it is the
greatest option available in the absence of documented experience. Numerous
intricate physiological and environmental elements play a role in determining
whether two plants will continue to grow or will be compatible when they are
connected through the asexual procedure of grafting. There are many levels of
compatibility or congeniality in grafting. A grafted apple on an oak tree dies
right away, but a grafted apple on a pear tree may grow well for a year or two

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before deteriorating and dying. Some lilacs grow on privet stocks for a period
of years before failing. In all other respects, the common apricot is the finest
stock for apricot variations, although in moist soils in cool climates, apricot
trees do better with some plum stocks than with apricot. Closely related plants
that may be successfully grafted have varying degrees of adaptation, which
makes it feasible for them to respond to soil conditions to a larger extent than
would otherwise be possible (Kumpers and Bishopp, 2015).
Types of grafts
Grafting can be categorised based on the area of the rootstock or different
parts of the plant's top where the scion is attached. There are three different
kinds of grafts: (1) detached scion graftage, which includes apical, side, bark,
and root graftage; (2) approach graftage, in which the root systems of the scion
and the rootstock are left in place until a successful graft union form; and (3)
repair graftage of mature trees. Detached scion graftage: A form of graft in
which a piece of the scion's shoot is cut off and grafted to the rootstock's apex
or side. Additionally, grafting roots uses it (root graftage). Approach graftage:
After a successful graft union has formed, the root system of the scion and the
shoot system of the rootstock are not removed. Repair graftage is the
application of graft to strengthen or repair damaged or frail trees (Kubota et
al., 2008; Lee and Oda, 2003; Legare, 2007; Melnyk and Meyerowitz, 2015;
Upchurch, 2006; Wang 2011).
1. Inarching or approach grafting

The distinctive characteristic of approach grafting is the joining of two

autonomously developing plants. The ability of the two plants to support
themselves independently ensures their survival even if the grafting procedure
fails for any reason. The fact that both plants are actively developing and there
is no time restriction for the healing of the graft union before the dependent
scion (top section) perishes from starvation considerably increases the
likelihood of success. As closely as you can without significantly harming the
established plant's root system, place an adapted, growing plant at the base of

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the non-adapted type. Shoots that are at least three-eighths of an inch in
diameter and preferably near to the same size should be tightly spaced from
both plants. A one to two-inch-long piece of bark is removed from both stems
at the location where the union is to take place. Each should have an identically
sized peeling area. The two peeling surfaces are then firmly bonded together
using electrical or budding tape. Wrap the region where the two peeling
portions are in touch fully with two coverings. Six to eight inches above the
graft union, trim part of the top leaves from the adapted variety. The grafted
union will heal more quickly as a result. In four weeks, the union should be
complete. The best time to execute this kind of grafting is during the growing
season. The remaining portion of the top of the native variety that has been
well-united (4 weeks or more) can be severed right above the graft union,
while the bottom or root system of the non-adapted, yellowing plant can be
severed just below the graft union. If the right rootstock was selected, the
issues with iron chlorosis and native soil pathogens have been resolved and
the graft union is now complete. If wilting happens due to insufficient root
system support, it could be required to lower the leaf area of the top of each
plant immediately after the piece is removed. Things will soon start to settle
down. The top, unadapted variety won't need to be severed from its own root
system if the sole issue has been a micronutrient (iron chlorosis) deficit; the
grafted, adapted variety root system will "feed" the sick plant what it need.
The vulnerable variety should be cut off from its root system and completely
reliant on the root system of the adapted variety, however, if the graft's goal is
to manage soil-borne illnesses.
2. Epicotyl (Stone) grafting

The epicotyl zone of the early seedlings is where this grafting technique
is applied, therefore the term epicotyl grafting. As rootstocks, mango seedlings

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are developed from the stones of young trees. During the months of July and
August, stones should be removed from local seedling mango trees of dwarf,
disease-free, and high producing kinds. In numerous methods of grafting,
proper scion selection and preparation are crucial. The scion should ideally be
a terminal, unflowered branch that is three to four months old. Prior to
detachment, selected scions should be defoliated on the mother plant for
around 7 to 10 days. On the chosen terminal shoot, a portion of the petiole that
corresponds to the rootstock's thickness should be left intact. The key to
successful stone grafting is matching scion thickness. Mango, cashew, etc.
choose a young seedling that is no older than 10 days and grow it in sized
polythene bags (15 cm X 22 cm). Remove the seedling's top section, leaving
a 5 to 6 cm long stalk (epicotyl). Make a vertical, downward incision (2-3 cm
long) along the middle of the remaining epicotyl using a sharp knife. As the
scion stick, use a terminal branch from a tested mother plant that is 3-4 months
old and is around 5-8 cm long. By making slanting and inward cuts of two to
three centimetres on opposing sides, the bottom end of the chosen scion is
chopped into a wedge shape. A wedge-shaped scion should be inserted into
the seedling's slit and securely fastened with tape or polythene strips.
Regularly mist the graft with water without getting the graft area wet. The
sprouting of the scion begins in around three weeks. If the seedlings are grown
in sand beds, they are dug out (with stones) 15 to 20 days after planting, when
they reach a height of 10-15 cm, and grafted as previously said. The grafted
seedling is then planted in polythene bags or pots to protect the stone and keep
the graft union above soil level. The optimal time to do epicotyl grafting is
from June to September.
3. Whip-and-tongue graft

Whip-and-tongue graft is particularly helpful for grafting relatively tiny

material with a diameter of 6 to 13 mm (1/4 to 1/2 in). If done correctly, it is
quite effective since there is significant vascular cambium contact, it heals

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rapidly, and it forms a strong union. The incisions made at the top of the
rootstock and the bottom of the scion must be identical. First, a 2.5-6 cm (1-2
1/2 in) long, smooth, sloping cut is created; longer cuts are done when working
with huge material. The tongues of the rootstock and scion are then placed
into one another. The vascular cambium layers must match on at least one side
and ideally both sides. This is crucial. For the same reason, it is best to avoid
using scions that are bigger than the rootstock. After the scion and rootstock
have been attached, they are firmly bound together using raffia, plastic
budding/grafting tape, or budding rubber strips. The tissues around the graft
union must not dry out, therefore until the graft union has formed, either the
graft union must be sealed with grafter's wax, Parafilm, or Buddy Tape, or the
plants must be kept in an environment with high relative humidity.
4. Splice graft (whip graft)

The second, or "tongue," cut is not made in either the rootstock or scion,
unlike the whip-and-tongue graft. Both the rootstock and the scion receive a
straightforward slanting cut with the same length and angle. As with the whip
graft, they are put together and wrapped or fastened. The vascular cambium
layers should correspond along the side of the rootstock where the scion is put
if it is smaller than the rootstock. When grafting plants with highly pithy stems
or wood that is too stiff to allow for a snug fit when a tongue is produced, such
in the whip-and-tongue graft, the splice graft is very helpful. The splicing graft
is used to graft disease-resistant rootstocks onto vegetable crops grown in
greenhouses. One Cotyledon Grafting is another name for this graft in relation
to vegetable crops like cucurbits or Solanaceae (OCG). While tying the splice
graft, the rootstock and scion need to be kept together. In field grafting, the

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cutting and tying must be done higher up on the rootstock since it is not
practical to utilise the technique at ground level. The grafter is thus responsible
for both tasks.
5. Cleft graft (split graft)

One of the first techniques for field grafting is the cleft or split graft. It is
used to topwork trees, either in the scaffold branches of a bigger tree or in the
little tree's trunk. This technique should only be used on rootstock branches of
topworking trees that are 2.5 to 10 cm (1 to 4 in) in diameter and of species
with relatively straight-grained wood that will break uniformly. Using a sharp
pruning saw, the limb that will be grafted or topworked is cut squarely. The
branch is then longitudinally broken in half using a chisel, a big knife, or a
specialised blade/wedge instrument made especially for cleft grafting. A 2 to
4-inch split is made in the limb, taking special care to place the split in the
centre of the limb. To prevent uneven splitting in species that don't split
uniformly, the initial cut might need to be done using a saw (termed saw kerf
grafting). The "cleft" is then pryed open and kept open using the wedge end
of the grafting tool or another appropriate implement once the split has been
created. The next step is to create a 3 to 4 bud scion stick between 4 and 6
inches in length for grafting into the cleft. The budstick should be made from
little limbs or water sprouts that had large, plump buds that were well
distributed and between 1/4 and 3/8 inches in diameter, indicating that they
had grown rapidly during the previous season of development. Avoid sprouts
with very big diameters as well as those that are thin, tiny, and have tightly
spaced buds. On precisely opposite sides of the stick, sloping cuts are
performed to trim the bottom end of the budstick. The budstick's incisions
should have a slope that nearly resembles the cleft's angle. The rootstock
cleft's form should be precisely replicated by the slope incisions on the

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budstick. To ensure optimum contact between the budstick and the rootstock
over its entire length, the incisions should be uniform in slope (and not wavy).
The quantity of touch will be insufficient to encourage the union's effective
healing if the budstick is too blunt. To encourage rapid healing, the cambia of
the two pieces must be perfectly aligned when the budstick is put into the cleft.
The thin line that separates the wood from the bark is known as the cambium.
The outside borders of the budstick and rootstock won't be flush since the bark
on the rootstock probably has a much thicker layer of bark than that on the
budstick. The two main criteria of success in cleft grafting are the capacity to
align the cambia of the two partners to be grafted and the capacity to maximise
the contact between the two parts to promote quick healing. The budsticks
should be held in place by the wood's inherent spring. The wedge keeping the
cleft open is gently withdrawn once both budsticks have been inserted and are
properly aligned. Grafting wax is applied to the cut ends of the budsticks, the
cut end of the rootstock, and the breaks in the cleft to keep the wood from
drying out.
6. Wedge graft (saw-kerf graft)

The diameter of the stock that has to be grafted is the same as for the cleft
graft: 5 to 10 cm (2 to 4 in). The scions need to be the same size as well: 10 to
13 cm (4 to 5 in) long and 10 to 13 mm (3/8 to 1/2 in) thick. To create a V-
wedge in the side of the stub, a sharp, hefty, short-bladed knife with a length
of about 5 cm (2 in) is employed. Two cuts are made, combining at the bottom
and separating by the width of the scion at the top. These incisions penetrate
the side of the stub approximately 2 cm (3/4 in) deep. The scion's base is cut
into a wedge form that matches the opening's size and shape perfectly. The
scion is tapped downward, firmly into position, slanting outward slightly at
the top, and matching the two vascular cambium layers so that the vascular
cambium layers cross. The scion should be secured in place so firmly that it

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would be challenging to remove if the incision is long enough and softly
tapered. Two scions should be put into a 5-cm wide stub spaced 180 degrees
apart, and three scions should be utilised, spaced 120 degrees apart, in a 10-
cm wide stub. All cut surfaces, including the tips of the scions, should be
completely waxed once all of the scions have been securely tapped into
position.
7. Saddle Graft

The scion and rootstock have to be the same size. On its two opposing
sides, the scion is prepared by making upward cuts through the bark and into
the wood. As the cuts are extended, the knife should cut deeper into the wood.
Before pulling back, the knife is pointed toward the centre of the scion piece,
where it progressively chisels away bits of wood to create the saddle shape.
For the rootstock to match the vascular cambium in the saddle of the scion,
the rootstock is sliced transversely and receives two upward incisions on either
side. To accommodate the saddle, the rootstock's apex has been cut. All
exposed cut surfaces must be sealed or kept in a grafting case until the graft
union has formed. The graft must also be knotted.

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8. Side Veneer grafting

Plant kinds that are difficult to root benefit greatly from side veneer
grafting. It employs miniature seedlings in pots as its rootstock. Compact and
dwarf conifers are the main beneficiaries of this sort of grafting. The season
before the grafting, the rootstock is cultivated in pots. It is then maintained in
the greenhouse for a few days to encourage the emergence of new roots.
During this time, the plant is not watered. Make a small incision at the stem's
base. Make an incision at the base of a scion that has the same diameter as the
rootstock. Use grafting tape or yarn to secure the cut section of the scion to
the cut surface of the rootstock. Apply wax or grafting paint to the whole
grafted region. Once the grafting has healed and fused together, take off the
tape or twine.
The side-veneer graft was once a common method for grafting difficult-
to-root camellia and rhododendron cultivars. It is now the most common
method of grafting conifers, particularly those with a compact or dwarf shape.
On potted rootstock, side-veneer grafting is frequently carried out. Getting the
Stock ready. The season before to grafting, rootstock is cultivated in pots, let
to become dormant, and then kept like other container nursery stock. The
rootstock is taken into a chilly greenhouse for a few days before to grafting in
order to promote fresh root development after being exposed to cold weather
for at least six weeks. Now is not the time to water the plant. At the base of
the stem on the potted rootstock, make a shallow downward incision that is
between 3⁄4 and 1 inch long to reveal a flap of bark with some wood still
attached. To remove the flap of bark and wood from the rootstock, make an
inward incision at the base. Getting ready for the Scion Select a scion whose
diameter is equal to or slightly less than that of the rootstock. At the base of
the scion, make a sloping incision that is between 3⁄4 and 1 inch long. Placing
the Scion in place Place the scion's cut surface up against the rootstock's cut

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surface. Make sure the cambia are in touch with one another. Using a rubber
grafting strip, tape, or grafting twine, secure the graft by holding the scion in
place. With heated grafting wax or grafting paint, the entire graft area is sealed.
After the union has healed, cut the rubber or twine. Never let the stem get
girdled by the binding substance.
9. Bark graft

Bark grafting is mostly used to top work fruit and flower plants. In
contrast to cleft grafting, this method may be used on rootstock with a diameter
of 4 to 12 inches. It is carried out in the early spring, just as the wood is
beginning to sap heavily. As with cleft grafting, the rootstock is cleanly cut
off with a sharp saw. Getting the Stock Ready Create a vertical incision
through the bark starting at the cut surface of the rootstock so that each scion
may be placed (2 inches long and spaced 1 inch apart). Getting the Scion ready
Prepare numerous scions for each graft as multiple scions are often put around
the cut surface of the rootstock. Each scion's base should only have one side
cut into a 112 to 2-inch tapering wedge. putting the Scion in. The scion should
be inserted with the bark somewhat looser, with the wedge-shaped tapering
surface of the scion resting on the exposed wood behind the flap of bark.
Replace the bark flap, push the scion firmly down behind it, and secure it in
place with one or two wire brads inserted through the bark and into the
rootstock. Every three to four inches along the cut edge of the rootstock, insert
a scion. Keeping the Graft Safe Grafting wax or grafting paint should be used
to cover all exposed surfaces. When the scions start to grow, only the strongest
one should be left on each stub; the others should be pruned away. Bark grafts
typically require staking or support during the first few years because they
frequently create weak unions.

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10. Bridge graft

Usually at or close to the base of the trunk, bridge grafting is used to

"bridge" a diseased or damaged region of a plant. Rodents, freezing
temperatures, disease organisms, or contact with grading or lawn maintenance
equipment are some typical causes of this type of damage. The bridge graft
acts as a pipeline for water and nutrients to pass through the injured region
while also providing stability. Bridge grafts are often performed in the early
spring, shortly before the start of vigorous plant development. They can be
carried out whenever the wounded plant's injured bark "slips." Getting the
Scion ready is Choose scions that are straight and roughly twice as long as the
damaged region that has to be bridged. At either end of the scion, make a 1⁄2
to 2 inch-long tapering cut on the same plane. Getting the Stock ready was to
ensure that the transplant is on healthy stems, remove any damaged tissue.
Create a flap of bark the same width as the scion and below the damaged area
on the rootstock. Avoid tearing the bark flap by gently folding the flap away
from the stock. Putting the Scion in the scion should first be inserted and
secured below the wound; press it under the flap with the cut end of the scion
resting on the wood of the hurt stem or trunk. Then repeat these techniques to
insert and secure the scion above the injury. Incorporate the scion tightly. As
with bark grafting, pull the flap over the scion and secure it in place. Insert the
scions with a small outward bend when grafting immature stems that could
sway in the wind. Over the injured region, bridge grafts should be placed
around 3 to 4 inches apart. Keeping the Graft Safe. With heated grafting wax
or grafting paint, all graft locations should be secured. Remove any buds or
shoots that appear on the scions both during and after the healing process.
Advantages of grafting
• Grafted trees bear fruit more quickly. A grafted tree will bear fruit in
about 2-4 years as opposed to 8-10 years for a tree developed from
seed.

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 • Fruit from a tree that was produced from seed may not taste very
good. Grafting is done to increase the fruit's size and flavour.
• It's possible that a tree developed from a seed won't bear fruit
identical to the tree from whence it was planted (mother tree).
However, a grafted tree will be just as excellent as the parent tree
from which it was taken (scion).
• For many years, a grafted tree will continue to produce fruit of the
same calibre.
• Grafting is the quickest method for producing good fruiting trees and
blooming shrubs in big quantities.
• Many plants with high economic value are difficult to grow through
conventional means of propagation, such as cutting and layering, but
they respond well to grafting.
• Although scions are extremely valuable kinds, they often have a
weak root system, reduced vigour, and inadequate defences against
diseases. By grafting them onto the stock, which is often a hardy,
wild relative of a scion, all these defects may be fixed.
• To retain the constancy of traits like size, colour, flavour, and other
characteristics, grafting is a method of cloning a scion variety.
• In response to a gardener's interest, many scion kinds can be grafted
onto a single stock plant; trees can also be given decorative forms,
etc.
• Fruit trees with grafts can be sold to provide revenue for the family.
• You save money by growing your own fruit and seedlings.
• Local production of seedlings reduces the amount of time needed to
choose the best fruit trees to plant.
• Some trees eventually become more resilient to illnesses and harmful
environmental factors than other trees. The rootstock (the plant being
grafted onto) imparts this disease resistance and hardiness to the
scion (the plant being propagated).
Disadvantages of grafting
• Costs Increased cost of using double seed, need for more greenhouse
space for transplanting double seedlings, use of certified seed, need
for skilled labour or additional training, and use of indeterminate

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 varieties rather than specific ones because they require less time to
produce before the investment is recouped.
• Poor graft implementation might stunt the growth of the plant and
cause physiological problems. Incompatibility that develops
throughout the transplant. The bottleneck that results from the
restriction of the xylem and phloem, which lessens the normal
movement of soil nutrients, is the principal expression of this issue
at the level of the union point of the graft-pattern.
• One of the biggest issues with grafts is incompatibility, which has a
detrimental effect on each person's genetic makeup.
• The following are examples of incompatibility:
o When the attempt to combine the volk and the pattern fails when
people die too soon.
o When the graft fails to mature properly or does not develop as
anticipated.
o When there is a noticeably disproportionate disparity between
the pattern's development and the graft's growth, or vice versa.
o At the time the bottleneck becomes apparent.
o When there is a diameter separation that is lower than the
combined diameter of the yolk and the pattern.
• The largest barrier to grafting is that it calls for knowledge and
expertise. He can't just decide to grow two batches of plants, graft
them together, and then leave them to develop on their own.
• Only when skilled employees are employed can grafting be
successful.
• This approach also requires a lot of time and work.
• Even so, the effectiveness of the grafting hinges on choosing plants
that are closely related.
• The grafting method can only only create a certain number of plants.
Conclusion
Modern horticulture depends on the traditional horticultural technique of
grafting. Long thought to be a strange technique and a dark secret, the practise
of fusing two live entities such that they subsequently work as one has long
been practised. This chapter explores the grafting, types, advantages and

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disadvantages for the crop improvement and upcoming developments in our
knowledge of and use of contemporary grafting. Grafting continues to be a
popular and necessary way of plant cloning for a wide range of uses and a
large range of species, despite recent advancements in propagation
technology, such as the use of rooting hormones, misting of cuttings, and
micro-propagation. Grafting is being employed for herbaceous vegetables in
addition to typical tree fruit crops and woody ornamentals. Reducing the use
of chemical pesticides for disease control is a major objective of contemporary
breeding of disease-resistant rootstocks, which helps to achieve the goal of
sustainable agricultural output. According to studies, the kind of rootstock
affects the production and quality characteristics of the scion fruit; as a result,
several seed firms are looking at grafting as a way to boost quality. It has been
claimed that grafting and the choice of rootstock can have an impact on pH,
flavour, sugar, colour, carotenoid content, and texture. Although opinions on
the benefits and drawbacks of grafting differ, it is generally accepted that for
the best fruit quality, the rootstock/scion combination must be carefully
chosen. Additionally, because many rootstocks have ideal temperature and
moisture ranges, it is crucial to examine rootstock/scion pairings under various
climatic and geographic situations.
References
1. Cantliffe, D.J. (2009). Plug transplant technology. Horticultural Reviews,
35, 397-436.
2. Goldschmidt, E.E. (2014). Plant grafting: new mechanisms, evolutionary
implications. Frontiers in plant Science, 5, 727.
3. Kubota, C., M.A. McClure, N. Kokalis- Burelle, M.G. Bausher, and E.N.
Rosskopf. 2008. Vegetable grafting: History, use, and current technology
status in North America. HortScience 43:1664-9.
4. Kumpers, B.M., & Bishopp, A. (2015). Plant grafting: making the right
connections. Current Biology, 25(10), R411-R413.
5. Lee, J.M., and M. Oda. 2003. Grafting of herbaceous vegetable and
ornamental crops. Hort. Rev. 28:61-124.
6. Legare, M. 2007. The future of grafting. Comb. Prod. Intl. Plant Proc.
Soc. 57:380-84.
7. Melnyk, C.W., & Meyerowitz, E.M. (2015). Plant grafting. Current
Biology, 25(5), R183-R188.
8. Mudge, K., Janick, J., Scofield, S., & Goldschmidt, E.E. (2009). A history
of grafting. Horticultural reviews. 5: 437-493.

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9. Upchurch, B.L. 2006. Grafting with care. Amer. Nurs. 203:18-22.
10. Wang, Y. (2011). Plant grafting and its application in biological research.
Chinese science bulletin, 56(33), 3511-3517.

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 Chapter - 12
Hybridization: Importance, Techniques and
Consequences

Authors
Manika Goswami
Department of Fruit Science, Dr. YS Parmar University of
Horticulture and Forestry, Nauni, Solan, Himachal Pradesh,
India
Sarita
Department of Fruit Science, Dr. YS Parmar University of
Horticulture and Forestry, Nauni, Solan, Himachal Pradesh,
India
Mohammad Ashiq Kuchay
Department of Fruit Science, Dr. YS Parmar University of
Horticulture and Forestry, Nauni, Solan, Himachal Pradesh,
India
Davinder
Department of Fruit Science, Dr. YS Parmar University of
Horticulture and Forestry, Nauni, Solan, Himachal Pradesh,
India

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Page | 186
 Chapter - 12
Hybridization: Importance, Techniques and Consequences
Manika Goswami, Sarita, Mohammad Ashiq Kuchay and Davinder

Abstract
Hybridization plays important role in the crop improvement. With the
advancement of breeding and biological sciences there is development of
various genomic tools and genomic analyses, it is very clear that gene flow
between diverged taxa generate distinct phenotypes. These phenotypes
allowed for adaptation in new environment and generate phenotypic diversity.
Hybridization have different consequences like heterosis, reinforcement,
hybrid speciation. These consequences affect the process of hybridization.
Hybridization results in the formation of new hybrids which are superior to
their parents for one of few characters. With the hybridization resistance can
be provided for different traits. Hybridization is the key step in the crop
improvement. Emasculation and pollination are the key steps in the
hybridization techniques. In this update, we discuss the consequences,
importance and techniques of hybridization in crop improvement.
Keywords: Emasculation, heterosis, hybrid speciation, hybridization,
reinforcement
1. Hybridization
Hybridization is the crossing between two genetically dissimilar parents
while hybridization among entities from diverse species of the same genus i.e.
intrageneric hybridization and two diverse genus of same family i.e.
intergeneric hybridization combinedly known as distant hybridization and
such crosses are known as distant crosses or wide crosses. Thomas Fairchild
was the pioneer who performed distant hybridization for the first time by
developing a hybrid by crossing Carnation (Dianthus caryophyllus) and Sweet
willian (Dianthus barbatus). First intergeneric hybrid, between Brassica
(cabbages) and Raphanus (radish) was given by Karpechenko leading to the
production of Raphanobrassica. Whereas Triticale was developed by crossing
wheat with rye possessing higher potential than raphanobrassica (Allard
1960).

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2. Importance of hybridization
2.1 Disease and insect resistance
Wide crosses can be used for transferring the disease resistance from wild
species to cultivated species (Harlan 1976; Knott and Dvorak 1976). The
hybridization can be done to impart resistance against Jassid in Cotton (Meyer
1974). In Okra with hybridization resistance to Yellow Mosaic Virus is
transferred from Abelmoschus manihot to Abelmoschus esculenta (Sandeep et
al., 2022). To provide resistance against leaf chewing insect in Groundnut the
hybridization has been performed in Arachis monticola and Arachis hypogeal
(War et al., 2012). In Tobacco resistance was imparted in against mosaic virus
from species N. repanda to N. tabaccum (Kumar et al., 2016). Likewise, in
Potato late blight and leaf roll resistance was provided by crossing Solanum
denissum with Solanum tuberosum (Yang et al., 2017). Fragaria chiloensis
crossed with Fragaria ananasa to provide resistance against aphids in
strawberry (Hancock et al., 2010).
2.2 Improvement in quality
Wide crosses can be used for improving the quality of cultivated plant
species. Some examples are given below (Simmonds 1979; Stalker 1980). In
cotton the character of fibre length and male sterility improved by crossing G.
thurberi with G. hisutum (Zang et al., 2021). Starch content increased in
Potato by crossing of Solanum acaule with Solanum tuberosum (Bradshaw et
al., 2006). While in tomato the carotenoid content increased by crossing of
wild species with cultivated tomato (Zhang and Stommel 2000). Sanghani and
Varu. In 2022 studied the effect of different species for the pollen on var Arka
Sahan and they found Annona squamosa as the best combiner and used further
for the yield and good taste.
2.3 Fruit with less seed
The first intergeneric hybrid has been produced between Citrus and
Citropsis with a smaller number of seeds. The less seed character transferred
from Citropsis to Citrus and the intergeneric hybrid so produced showed
resistance to many diseases,
2.4 Abiotic stress tolerance
To develop fruit free from fruit drop in mango, different species of Mango
has been used in the experiment including Mangifera indica, Mangifera
zeylanica and Mangifera odorata among which Mangifera odorata has been
found best when crossed with Arka Anmol recored highest fruit set and less
fruit drop (Dinesh et al., 2016).

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3. Techniques/Procedure of hybrid cization
3.1 Choice of parents
3.2 Evalution of parents
3.3 Emasculation
3.3.1 Hand emasculation
3.3.2 Suction method
3.3.3 Hot water emasculation
3.3.4 Cold treatement
3.3.5 Chemical emasculation
3.4 Bagging
3.5 Tagging
3.6 Pollination
3.7 Harvesting and storing of F1 seeds
3.1 Choice of parents: The choice of parents depends upon the breeding
objective. Increased yield, insect pest resistance and now a days resistance
to abiotic stresses are the major breeding objectives of the breeder. Hence
the selection of parents should be done on the basis of the purpose of the
hybridization. The parents should be well adapted to new environment
and can be easily mated while crossing.
3.2 Evaluation of parents: The evaluation of parents is the necessary step
for the breeding programme as the performance of parents should be done
for the character which is to be transferred.
3.3 Emasculation: Removal of male part or stamens or anthers or removing
or killing of pollen grains without affecting the female part of the plant is
called as emasculation. The main objective of emasculation is the
prevention of self fertilization and promotion of cross feritilization or
cross pollnation. If the male part of the flower remains there, then there is
chance of self pollination and this is the case in Peach where self
pollination occurs because the flower of peach is hermaphrodite or pefect
flower which carries both the reproductive organ in the same flower and
hence self pollination is the rule.
3.3.1 Hand emasculation: This process of emasculation, performed in
those species which have large size of flowers and in which
stamens or anthers can be removed easily with the help of foreceps.
Emaculation should be done before the anthers are mature and
before the stigma become receptive.

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 3.3.2 Suction method: This method of emasculation is performed in the
species which have small size of flowers, the emasculation should
be done in the morning hours and the petals are removes as the
removal of stamens is difficult.
3.3.3 Hot water emasculation: The pollen grains are more sensitive to
environment as well as the high temperature and hence the pollen
grains are subjected to high temperature by treating them with hot
water in order to kill the pollen grains. The time and temperature
of pollen treatment varies from species to species.
3.3.4 Cold treatment: Like hot water, the cold water treatement also
kills pollen grains without damaging gynoecium.
3.3.5 Chemical emasculation: Chemicals like GA3, Ethephon used for
chemical emasculation.
3.4 Bagging: After emasculation the flowers or the inflorescence are
enclosed in suitable bags of appropriate size to prevent cross pollination.
The bags used for bagging are made up of butter paper or fine cloth.
3.5 Tagging: The emasculated flowers are tagged just after bagging with
tags. The tags are available in different sizes and made up of different
material. The circular tags are 3 cm in diameter while the rectangular tags
are 3 X 2 cm.
3.6 Pollination: Pollination along with emasculation are the two main steps
which determine the amount of seed set in hybridization. In pollination,
the anthers which dehiscenced and release pollen grains were placed on
female reproductive organ and this process is called as pollination.
3.7 Harvesting and storing of F1 seeds: The crossed fruit or pods or heads
harvested and seeds were dried and stored for further use. The seeds from
each cross should be stored separately so that mixing should not be done.
4. Consequences of hybridization
Hybridization affects the evolutionary trajectory of lineages. Koelreuter
in 1766 observed hybrid vigour as a consequence of hybridization as the
interspecific hybrids are mostly sterile and difficult to produce. Hybrids are
sterile, inviable or very rare as the exchange of genetic material through
different species is not easy as well as possible. Hybridization with gene flow
has many consequences as compared to the hybridization without gene flow.
Therefore, different consequences of hybridization as follow:

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4.1 Heterosis
Hybrids can be generated by crossing of two genotypes which are similar
or of different ancestory after controlled pollination and the hybrid so obtained
is superior to both the parents. This process in plants is known as hybrid vigour
or heterosis. Koelreuter in 1766 and Darwin in 1876 found the observation
and revealed in the experiments the process of heterosis but none of them
explained the mechanism behind it (Mayr, 1986; Chen, 2013). Then in 1908
and 1911 Shull determine the genetic mechanism of heterosis in maize. He
gave two competing hypothesis i.e., Dominance (Jones, 1917) and
overdominance hypothesis (East 1936). According to the dominant
hypothesis, recessive alleles showed their position at different loci and these
recessive alleles masked by beneficial allele from distinct parents. While on
the other note, overdominance hypothesis states that homozygous is inferior
to heterozygous genotypes (Chen, 2013). Early workers found that heterosis
have a single cause (Crow, 1948) but later it was found that heterosis
contribute multiple causes (Grant, 1975; Kaeppler 2012). QTL (Quantitative
trait loci) has been used to characterize different loci which contribute to
heterosis. These findings aim that heterosis result of many loci which is a
combination of dominant, overdominant and epistatic effects (Tang et al.,
2010; Zhou et al., 2012; Shen et al., 2014; Shang et al., 2015). Krieger et al
in 2010 studied that in tomato, wild types are heterozygous are are superior
than homozygous genotypes having single flower trus. Hence from this study
it is revealed that yield is governed by overdominance at single locus. Studies
done by Groszmann et al in 2013 and Greaves et al in 2015 showed that
interaction between epigenetic systems in genotypes showed high significance
of heterosis.
4.2 Transgressive segregation
Trangressive segregation defined as the hybrid production which exceeds
the parental phenotypes. It is distinct from heterosis as it is evident in the 2 nd
filial generation i.e., F2 and in the later stages it stable itself, if established
(Rieseberg et al., 1999). Transgressive segregation is most commonly present
in hybrids. Stebbins in 1950 studied that the traits showing transgressive
segregation adapted to new environment easily Lotsy et al (1916); Hagedoorn
and Hagedoorn-Vorstheuvel La Brand (1921). The hypothesis has been
proposed (Rieseberg et al., 1999), but the mechanism which is genetically
controlling the transgressive segregation are epistatic action and
complementary gene action (Rieseberg et al., 1999; Dittrich-Reed and
Fitzpatrick, 2013). In case of former, both the parents have additive alleles of
different/opposite signs while for later, both the alleles have similar signs. In
recent studies, a new mechanism discovered for this phenomenon i.e., small

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interfering RNAs (Shivaprasad et al., 2012). QTLs have also been studied for
complementary gene action and epistatis or both (Hagiwara et al., 2006; Mao
et al., 2011).
4.3 Adaptive introgression
The gene flow between parents has been sustained by the production of
hybrids. The hybrids so obtained are fertile as well as superior and hence they
used in backcross for further breeding programme and this is called as
introgression (Levin et al., 1996; Todesco et al., 2016). The introgression
produces new, adaptive genetic variation in a population. The idea of
introgression given by Johannes Lotsy while the term coined by Edgar
Anderson (Anderson and Hubricht, 1938). Anderson revealed that
introgression produces large variation in a population, which move from one
generation to another into a new ecological niche. Due to limitation of
indentification of loci which are introgresses, there is limitation to study
adaptive introgression. However, in Tradescantia, Melandrium and
Helianthus the introgressed loci has been studied (Anderson, 1949; Stebbins,
1950). For the study of adaptive introgression, variation has been produced
when there is gene flow in populations and this variant is adaptive. Different
techniques which measure selection on traits namely reciprocal transplant
experiments and common garden experiments.
4.4 Reinforcement
Hybridization among diverged lineages often is not progressive. To
increase the hybrid fertility, the selection should not be done from the closely
related ancestory. Reinforcement is the process of increasing the reproduction
isolation which is due to selection with the main aim to decrease hybridization.
Reinforcement in plants has been reviewed by Hopkins in 2013, but the
historical importance of reinforcement in hybridization is reviewed here.
Reinforcement is a process where natural selection favours by increasing the
reproductive isolation between two species. The idea was developed by Alfred
Russel Wallace and also called as Wallace effect (Grant, 1966). However, the
reinforcement has been done mostly in animal system (Dobzhansky and
Koller, 1938; Blair, 1955). Reinforcement means mating between closely
related ancestors. But this cause low viability as well as low fertility. Costly
hybridization favours selection ehich focuses on new traits that increase
assortative mating. As the gene flow in hybridization, prevents the
reproductive isolation by which reinforcement use become controversial
among the species that occur at same place at same time (Felsenstein, 1981;
Butlin, 1987). Hence, hybridization is success a well as major hinder of
reinforcement (Kirkpatrick, 2000). New alleles have to be genetically

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associated with allele which causes sterility in order to confer assortative
mating (Servedio, 2009). Extensive theoretical research has showed that the
viability of reinforcement is simply a balance between the evolutionary forces
of selection, gene flow and recombination (Liou and Price, 1994).
4.5 Hybrid speciation
Hybridization is widespread, but the evolution of a unique, isolated hybrid
lineage is very few. A new hybrid lineage developed from allopolyploidy or
from homoploid hybrid speciation. The former lineage produced by the fusion
of unreduced gametes while the later formed hybrid lineage without a change
in ploidy (Ramsey and Schemske, 1998). Allopolyploid hybrid speciation is
the more feasible and commonly occurring type of hybrid speciation (Soltis
and Soltis, 2009). A review reveled by Barker et al 2016 stated that 11% of
species among 47 plant genera derived from allopolyploid origin. However
very less examples of homoploid hybrid speciation were recorded (Mavarez
et al., 2006; Lukhtanov et al., 2015). Because of the scarcity of examples,
researchers revealed that hybridization were more likely in stable hybrid zones
than homoploid hybrid speciation (Buerkle et al., 2003). Furthermore, studies
support that the genetic divergence affects the relative likelihood of
homoploid (more likely between less-diverged species) or polyploid (more
likely between more-diverged species) hybrid speciation (Chapman and
Burke, 2007). This is similar with the findings that homologous chromosomes
will pair with greater accuracy as the genetic divergence among the
progenitors of an allopolyploid increases which leads to higher fertility
(Stebbins, 1947).
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 Chapter - 13
Role and Importance of ICT in Agricultural
Extension

Authors
Shruti Samadder
Ph.D. Research Scholar, Department of Extension Education
(PGCA), Dr. Rajendra Prasad Central Agricultural University,
Pusa, Samastipur, Bihar, India
Deepak Rao
Ph.D. Research Scholar, Division of Seed Science and
Technology, IARI, New Delhi, India

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Page | 200
 Chapter - 13
Role and Importance of ICT in Agricultural Extension
Shruti Samadder and Deepak Rao

Abstract
Information and Communication Technology (ICT) has the potential to
revolutionize the way agricultural extension services are delivered. The use of
ICT in agricultural extension can help to improve the efficiency, effectiveness,
and reach of extension services, and can also help to empower farmers with
the information and tools they need to improve their livelihoods. ICT can
provide farmers with access to real-time information on weather, market
prices, and crop management, which can help them make more informed
decisions about their crops and increase their productivity. It can also provide
them with access to financial services such as microfinance and insurance,
which can help them invest in their farms and manage risk. ICT can also
improve connectivity between farmers, extension workers, and researchers,
enabling them to share information and knowledge more easily, and also
access to value-added services such as e-commerce, telemedicine, and e-
governance services. However, to be successful, ICT projects for agricultural
extension must be targeted at the right audience, use appropriate technologies,
and have effective communication and dissemination strategies in place. It is
also important to ensure that the projects are owned and managed by the local
community, with participation from farmers, extension workers, and other
stakeholders. Overall, the use of ICT in agricultural extension has the potential
to greatly enhance the effectiveness and efficiency of extension services, as
well as empower farmers with the information and tools they need to improve
their livelihoods and increase agricultural productivity.
Introduction
India is an agrarian country. The primary source of income for the society
is agriculture and its allied sectors. To increase the pace of the development
in sector whether its agriculture or industry ICT is one of the important
catalysts. Information and communication technology, or ICT, is occasionally
referred to as IT (Information Technology). The phrase is generally accepted
to refer to all tools, networking components, software, and systems that let

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people and organisations interact online (i.e., companies, non-profit
organizations, governments and criminal enterprises). Technology used to
transfer, process, store, produce, display, disseminate, or exchange
information through electronic methods is referred to as information and
communication technology service. ICT tools are the equipment used to offer
clients services. The ICT tools include smartphones, televisions, FM radios,
the internet, call centres, multimedia CDs, movies, computers, the World
Wide Web, and Google. ICT is one of the pillars of economic development to
increase national competitiveness. It can raise awareness of essential, practical
concerns including social and health issues and serve as a tool for learning and
education. As a result, it can enhance the quality of human existence.
Problemes in agiculture
Agriculture is flourishing in last few decades but though there are few
problems that this sector is facing many challenges regarding sustainable
development, seasonal unemployment, market issues, land problem, climate
change, nutrition problem.
Sustainable development
Sustainable issues related to agriculture are deforestation, soil
degradation, excessive amount of pesticide and fertilizer use, water scarcity
climate change.
Seasonal unemployment
In India agriculture is still highly dependent upon weather and seasonal
change. It is a phenomenon where farm workers are only employed during
certain times of the year such as during planting or harvesting seasons.
Market issues
Various issues related to market are also there. Sometimes farmers facing
challenges in choosing between market relevant high profit crop and
traditional crop and sometimes they did not receive the bare minimum price
for their crop.
Land related problem
In India most of the farmers have fragmented land and are under farming
is decreasing continuously so pressure over per unit of land is increasing
continuously.
ICT scenario in India
According to Statista in India around 784 million people were using

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smartphones (Statista, 2020). In one study MNC Deloitte reveal that by the
end of 2026 India smartphone users will become 1 billion and India will
become 2nd largest smartphone user country (Deloitte,2022).
Infrastructure of ICT in India
Information and Communication Technology has growing continuously
and significantly in recent years, and to boost it govt. had already taken various
initiatives and policies to increase the people e-readiness.
Telecommunication: Telecommunication is one of the major sectors of
ICT. 784 million people were using smartphones (Statistia, 2020). 622 million
active internet users around the country (Statista, 2022). To make internet
more accessible to rural India govt. launched Bharatnet which is world
largest broadband project that provide broadband connectivity over 2.5 lakhs
gram panchayat (BBNL, 2023).
Data centre: National Informatics Centre, Ministry of Electronics and
Information Technology servers as a centralized repository for government
data and services. It is the technology partner, helps is designing and
developing govt. softwires, Investigate and provide guidance on using
emerging technologies and provide infrastructure to the ICT.
E-government services: The Indian Government has also made a lot of
effort to connect its citizens to the digital world and encourages and facilitates
the use of e-government services for easy and quick access for citizens, it has
implemented Digital India Program which has the vision of transforming
India into a digitally empowered society and knowledge economy.
ICT in agricultural extension
Extension always plays an important role in transfer of knowledge from
experts to farmers and other stake holders of the agriculture. Extension
services provide farmers best practices, and scientific advancement, that
improve crop yield, reduce cost and protect the environment. ICT help
extension to fulfil the goal.
Information and communication technology (ICT) can play a significant
role in agricultural extension, which is the transfer of knowledge and
technology to farmers and other stakeholders in the agriculture sector. Some
of the ways in which ICT can support agricultural extension include:
Improving access to information: ICT can help to make a wide range
of information and resources available to farmers and extension workers, such
as weather forecasts, market prices, and best practices for crop management.

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 • Online platforms: ICTs like the internet and mobile phones provide
access to a vast amount of information on a wide range of agricultural
topics, including best practices, new technologies, and market prices.
• Agricultural databases: ICTs can be used to create and manage
databases of information, such as weather data, soil maps, and crop
yield records. These databases can be accessed by farmers, extension
agents, and researchers to help inform their decision making.
• Video and multimedia content: ICTs like smartphones and tablets
can be used to create and share video and multimedia content, such
as instructional videos, podcasts, and webinars, which can provide
farmers with access to information in an engaging and accessible
format.
• SMS and mobile apps: ICTs can also be used to send information
directly to farmers' mobile phones through SMS or mobile apps,
which can provide them with timely and relevant information in areas
with limited internet connectivity.
• Overall, the use of ICTs can help to increase the accessibility and
availability of information to farmers, which in turn can help to
improve their productivity and livelihoods.
Enhancing communication and collaboration: ICT tools, such as
mobile phones and social media platforms, can facilitate communication and
collaboration among extension workers, farmers, and other stakeholders. This
can help to improve the sharing of knowledge and expertise and facilitate the
adoption of new technologies and practices.
• Mobile phone and internet-based technologies: These can be used
to connect extension agents and farmers, allowing them to share
information, ask questions, and receive feedback in real-time.
• Social media platforms: These can be used to create online
communities of farmers and extension agents, where they can share
information, collaborate on projects, and learn from each other.
• Geographic Information Systems (GIS): These can be used to map
agricultural resources and identify potential problem areas, which
can aid extension agents in providing targeted information to
farmers.
• Remote sensing: These can be used to gather data on crop health and
soil conditions, which can be used to inform farmers about the best
time to plant, fertilize, and harvest their crops.

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 Overall, the use of ICT in agricultural extension has the potential to
improve the effectiveness and efficiency of extension services, ultimately
resulting in more productive and sustainable agriculture.
Supporting distance education and training: ICT can be used to deliver
extension services remotely, using platforms such as online courses, webinars,
and video conferencing. This can be particularly useful in areas where
extension workers are scarce or hard to reach.
Some examples of how ICT can be used to support distance education in
agricultural extension include:
• Online courses and webinars: ICTs like the internet and video
conferencing tools can be used to deliver educational content to
farmers remotely, allowing them to learn at their own pace and on
their own schedule.
• Mobile learning: ICTs like smartphones and tablets can be used to
deliver educational content to farmers in the form of mobile apps and
SMS-based services, which can be accessed even in areas with
limited internet connectivity.
• Interactive multimedia: ICTs can be used to create interactive
multimedia content, such as videos and animations, which can be
used to explain complex concepts and procedures in an engaging and
accessible format.
• Virtual reality and simulations: ICTs can be used to create virtual
reality and simulation-based training programs, which can provide
farmers with an immersive learning experience and the ability to
practice new skills in a safe and controlled environment.
Overall, the use of ICT in distance education in agricultural extension can
help to increase access to educational resources for farmers, especially those
living in remote or underserved areas. It also allows farmers to learn at their
own pace and convenience, which is particularly beneficial for farmers with
busy schedules or limited mobility.
Facilitating data collection and analysis: ICT tools, such as sensors and
drones, can be used to collect data on crops, soil, and weather conditions. This
data can then be analysed to inform extension recommendations and support
decision-making.
Overall, the use of ICT in agricultural extension can help to improve the
efficiency and effectiveness of extension services, as well as increase the
uptake of new technologies and practices by farmers.

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Policies regarding ICT in agricultural extension
Policies regarding the use of ICT (Information and Communication
Technologies) in agricultural extension can vary depending on the country and
region. However, some common elements of policies that support the
integration of ICT in agricultural extension include:
Investment in infrastructure: Governments may invest in the
development and maintenance of infrastructure such as internet connectivity,
telecommunication.
Increasing access to ICT infrastructure and services: This includes
policies to improve internet access and mobile phone coverage in rural areas,
as well as the development of agricultural databases and other digital
resources.
Promoting the use of ICT in extension services: This includes policies
to encourage extension agents to use ICT tools and resources in their work, as
well as the development of training programs to help them acquire the
necessary skills.
Encouraging innovation in ICT for agriculture: This includes policies
to support the development of new ICT tools and technologies that can be used
to improve extension services, as well as the creation of incentives for
companies and organizations to invest in this area.
Building capacity of farmers and other stakeholders in the use of
ICT: This includes policies to provide training and support to farmers and
other stakeholders to help them acquire the necessary skills and knowledge to
use ICT in their farming activities.
In India, the government has implemented several policies and
initiatives to promote the use of ICT in agricultural extension. Some
examples include:
• National e-Governance Plan (NeGP) for Agriculture: This
initiative aims to create a comprehensive e-governance framework
for the agriculture sector, with a focus on providing farmers with
access to information and services through a variety of ICT channels,
including the internet, mobile phones, and Common Service Centers
(CSCs).
• National Agriculture Market (e-NAM): This is an electronic
trading portal that connects farmers to markets across the country,
providing them with real-time information on market prices and
trends, and enabling them to sell their produce directly to buyers.

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 • Krishi Vigyan Kendras (KVKs): These are rural centres that
provide farmers with training and support in the use of modern
technologies and practices. The KVKs have been provided with ICT
tools such as computer, internet and other necessary facilities to
provide the farmers with the latest information and technologies.
• National Centre for Precision Agriculture (NCPA): This centre
focuses on developing and promoting precision agriculture
technologies and practices, using ICTs such as GIS, Remote Sensing,
and precision farming equipment.
• Digital India Initiative: This initiative aims to provide all citizens
with access to digital services, including e-governance, e-commerce,
and e-education, and it also includes the development of a digital
infrastructure that can support the agriculture sector.
Various initiatives on ICT projects

S. No. Name Description

It is a method of problem-solving intended to address both
little and major issues faced by Indian farmers. Depending
aAQUA
on the difficulty, responses to your Agri-related questions
(Almost all
are answered in 24 to 72 hours. Experts work for their
1. question
respective companies and provide their services for free.
answered)
Language: Hindi, Marathi, English
Collaboration of IIT Bombay, KVK, Vigyan Ashram
(Dept. of Science and Technology, Govt. of India)
Digital Green is a non-profit organization that uses digital
technology to improve the lives of smallholder farmers in
developing countries. The organization creates videos that
demonstrate best practices for farming and other rural
livelihood activities, and then disseminates the videos
through community networks to reach farmers who may not
2. Digital Green have access to traditional forms of extension services. The
organization also provides training and support to local
partners to help them create and disseminate their own
videos. Digital Green has been implemented in many
countries like India, Ethiopia, Ghana, Nigeria, and has
impacted many farmers livelihoods.
(Sarvan Raj, 2007)
By enhancing access to information and knowledge sharing,
ICT-based agricultural education has the potential to
e-Arik (e-
3. strengthen farming communities. One such project that
Agriculture)
makes use of ICT as a teaching tool is the Village
Knowledge Centres in North East India.
eSagu is an online platform that aims to provide real-time
4. e-Sagu
information to farmers about the market prices of various

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 crops, weather forecast, and other agricultural-related
information. The platform also aims to connect farmers with
buyers, agri-input dealers and other service providers to
ensure better prices for their produce and timely availability
of inputs. The website also has information on various
government schemes, policies, and programs related to
agriculture.
Place: Telengana
VASAT (Virtual Academy for the Semi-Arid Tropics) is an
online platform that provides training and education on
sustainable land management in semi-arid regions. The
platform aims to empower farmers and other stakeholders in
dryland areas with the knowledge and tools they need to
improve their livelihoods and adapt to the challenges of
5. VASAT
climate change. VASAT offers a range of educational
resources, including videos, interactive simulations, and
quizzes, as well as opportunities for online collaboration and
networking. The program is developed by the International
Crops Research Institute for the Semi-Arid Tropics
(ICRISAT) in collaboration with various partners.
The Soil Health Card (SHC) program is a government
initiative in India to provide farmers with information about
the health of their soil. The program aims to help farmers
improve crop yields and soil fertility by providing them with
regular soil test results and recommendations for nutrient
management. E-Extension is a digital platform for the Soil
e-Extension
Health Card program, which allows farmers to access their
(Soil health
6. soil test results and recommendations electronically, rather
card
than receiving a physical card. The farmers can access their
programme)
soil test results, recommendations for nutrient management
and also get information about various other schemes and
programmes through the e-Extension portal. This enables
farmers to access their soil information at any time and from
any location, improving the efficiency and accessibility of
the program.
A Village Resource Centre (VRC) is a community-based
facility that provides rural residents with access to
information and services related to agriculture, health, and
livelihoods. The VRCs aim to bridge the gap between rural
communities and the resources they need to improve their
livelihoods. They typically provide a wide range of services,
including training on sustainable agriculture and livelihoods,
ISRO-Village
7. access to information on agricultural practices, health, and
resource centre
other topics, and support for farmers' groups and
community-based organizations. VRCs also serve as a
platform for farmers to share their experiences and
knowledge, and for extension workers and researchers to test
and disseminate new technologies and practices. The goal of
VRC is to empower rural communities by providing them
with the information and services they need to improve their

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 livelihoods and adapt to the challenges of a changing
environment
The M. S. Swaminathan Research Foundation (MSSRF) is a
non-profit organization in India that works towards
sustainable agricultural development and rural livelihoods.
The Village Knowledge Center (VKC) is one of the
programs run by the MSSRF. The Village Knowledge
Centres (VKCs) are a network of rural resource centres that
provide farmers and rural communities with access to
information and services related to agriculture, health, and
livelihoods.
VKCs are set up in villages and aims to bridge the gap
between research and rural communities by providing
8. MSSRF
farmers with access to information and services related to
agriculture, health, and livelihoods. The centers provide a
range of services including access to information on
agriculture, health, and livelihoods; training on sustainable
agriculture and livelihoods; and support for farmers' groups
and community-based organizations. The VKC also serves
as a platform for farmers to share their experiences and
knowledge. This program aims to empower rural
communities by providing them with the information and
services they need to improve their livelihoods and adapt to
the challenges of a changing environment.
The Warana Wired Village Project is a rural development
initiative in India that aimed to provide internet connectivity
and digital services to the Warana valley region of
Maharashtra state. The project was launched in 2000 by the
Warana Co-operative Sugar Factory Ltd. in collaboration
with the Government of Maharashtra and various other
partners.
The project aimed to provide internet connectivity and
digital services to the Warana valley region of Maharashtra
Warana Wired
9. state and also to provide training to the farmers and rural
Village Project
citizens in the use of computers and the internet. The project
also aimed to bridge the digital divide between urban and
rural areas, and to provide access to information and services
related to agriculture, health, education, and livelihoods. The
project also aimed to promote e-commerce and telemedicine
services in the region. The goal of the project was to
empower rural communities by providing them with access
to information and services that would improve their
livelihoods and standard of living
DACNET (Digital Advancement of Agriculture in India
Network) is an e-governance project of the Government of
India that aims to provide farmers and other stakeholders in
10. DACNET the agriculture sector with access to information and services
related to agriculture. The project is implemented by the
Department of Agriculture, Cooperation, and Farmers
Welfare (DACFW) under the Ministry of Agriculture and

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 Farmers Welfare.
The project aims to provide a wide range of services to
farmers, including access to information on weather, market
prices, and other topics; training on sustainable agriculture
and livelihoods; and support for farmers' groups and
community-based organizations. DACNET also provides
services to the department of agriculture such as online
submission of crop cutting experiment data, online
distribution of fertilizers, pesticides and seeds, and online
monitoring of various schemes and programmes. The goal of
the project is to empower farmers and other stakeholders in
the agriculture sector by providing them with the information
and services they need to improve their livelihoods and adapt
to the changing environment.
ICT-e-Choupal is an e-governance initiative in India that
uses Information and Communication Technology (ICT) to
connect farmers in rural areas with markets, information, and
services. The initiative was launched in 2000 by the Indian
multinational corporation, ITC Limited (formerly known as
Indian Tobacco Company).
The e-Choupal initiative uses ICT to provide farmers with
access to information on weather, market prices, and
agricultural best practices, as well as to connect them directly
with buyers. This enables farmers to make more informed
decisions about what to grow and when to sell, which in turn
11. ICT-e-Choupal improves their income and livelihoods. The e-Choupals also
provide a platform for farmers to share information and
experiences, and for extension workers and researchers to
test and disseminate new technologies and practices.
The e-Choupals consist of a network of rural kiosks
equipped with computers and internet connectivity, where
farmers can access information and services. The kiosks are
typically located in rural areas and managed by trained local
entrepreneurs. The e-Choupals have been implemented in
many states in India, and have been recognized for their
potential to improve rural livelihoods and bridge the digital
divide between urban and rural areas.
Ishakti is an e-governance project in India that aims to
provide farmers with access to information and services
related to agriculture, health, and livelihoods. The project is
implemented by the M. S. Swaminathan Research
Foundation (MSSRF) and is supported by the Government
of India.
iShakti The project uses ICT (Information and Communication
Technology) tools such as mobile phones, tablets, and
computers to deliver information and services to farmers.
The project aims to provide farmers with access to real-time
market prices, weather forecasts, and other information that
can help them make informed decisions about their crops.
The project also aims to provide farmers with access to

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 financial services, such as microfinance and insurance, and
to connect them with buyers and suppliers.
iShakti is an innovative way of providing extension services
to farmers by leveraging the power of technology and mobile
phones. By providing farmers with access to real-time
information and services, the project aims to improve their
livelihoods and promote sustainable agricultural
development
IFFCO Kisan Sanchar Limited (IKSL) is a subsidiary of
Indian Farmers Fertilizer Cooperative Limited (IFFCO), one
of the largest cooperative societies in India. IKSL provides
mobile-based services to farmers to help improve their
livelihoods and increase agricultural productivity.
IKSL's services include providing farmers with information
on weather, market prices, and crop management, as well as
access to financial services such as microfinance and
insurance. The company also offers a variety of value-added
IFFCO Kisan services such as e-commerce, telemedicine, and e-
Sanchar Ltd. governance services. Through its mobile platform, farmers
can also connect with buyers and suppliers, and access a
range of other services such as soil testing and crop
insurance.
IKSL's mission is to empower farmers with information and
services that can help them improve their livelihoods and
increase agricultural productivity. Through its mobile
platform, farmers can access a range of services that can help
them make more informed decisions and increase their
income.

ICT initiatives by the National Agricultural Innovation Project (NAIP)

by ICAR
The National Agricultural Innovation Project (NAIP) is a program of the
Indian Council of Agricultural Research (ICAR) that aims to promote
agricultural research and innovation in India. The program focuses on
developing new technologies and practices that can help farmers increase
productivity and improve their livelihoods.
NAIP has implemented several ICT (Information and Communication
Technology) initiatives to promote agricultural research and innovation. Some
of these initiatives include:
e-Krishi: This is an online platform that provides farmers with access to
information and services related to agriculture, health, and livelihoods. The
platform includes information on weather, market prices, and crop
management, as well as access to financial services such as microfinance and
insurance.

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 Agri-cloud: This is an online platform that enables farmers to access a
range of services including weather forecasts, soil testing, and crop insurance.
The platform also allows farmers to connect with buyers and suppliers, and
access a range of other services such as e-commerce and telemedicine.
Krishi Vigyan Kendra (KVK) Network: This is a network of rural
resource centers that provide farmers and rural communities with access to
information and services related to agriculture, health, and livelihoods. The
KVKs provide a range of services including training on sustainable agriculture
and livelihoods, access to information on agricultural practices, health, and
other topics, and support for farmers' groups and community-based
organizations.
mKrishi: This is a mobile-based service that provides farmers with
information on weather, market prices, and crop management. The service
also includes a range of value-added services such as e-commerce,
telemedicine, and e-governance services.
These initiatives by NAIP and ICAR aims to empower farmers with
information and services that can help them improve their livelihoods and
increase agricultural productivity. By providing farmers with access to real-
time information and services, the project aims to improve their livelihoods
and promote sustainable agricultural development.
Impact of ICT initiatives
The impact of ICT (Information and Communication Technology)
projects on agriculture and rural development can be significant. Some of the
potential benefits include:
Improved access to information: ICT projects can provide farmers with
access to real-time information on weather, market prices, and crop
management, which can help them make more informed decisions about their
crops. This can lead to increased productivity and improved livelihoods.
Increased access to financial services: ICT projects can also provide
farmers with access to financial services such as microfinance and insurance,
which can help them invest in their farms and manage risk.
Improved connectivity: ICT projects can improve connectivity between
farmers, extension workers, and researchers, enabling them to share
information and knowledge more easily. This can help farmers access the
latest research and technologies, and can also help researchers test and
disseminate new technologies and practices.
Increased access to value-added services: ICT projects can provide
farmers with access to a wide range of value-added services such as e-

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commerce, telemedicine, and e-governance services. This can help farmers
access new markets, improve their health and wellbeing, and access
government services more easily.
Improved efficiency: ICT projects can help to improve the efficiency of
agricultural supply chains by enabling farmers to connect with buyers and
suppliers more easily. This can help to increase the amount of produce that
reaches the market and can also help to improve the prices that farmers receive
for their crops.
However, it's important to note that the impact of ICT projects on
agriculture and rural development will depend on many factors such as the
type of ICT project, the target population, infrastructure and resources
available, and the level of adoption and usage.
Conclusion
There are several lessons that can be learned from the use of ICT
(Information and Communication Technology) for agricultural extension.
Some of these include:
The importance of targeting the right audience: To be successful, ICT
projects for agricultural extension should be targeted at the right audience.
This means identifying the farmers and rural communities that are most in
need of the information and services being provided and tailoring the project
to meet their specific needs.
The need for local ownership and participation: ICT projects for
agricultural extension should be owned and managed by the local community,
with participation from farmers, extension workers, and other stakeholders.
This can help to ensure that the project is responsive to local needs and is
sustainable in the long term.
The importance of using appropriate technologies: To be effective,
ICT projects for agricultural extension should use technologies that are
appropriate for the target audience and the local environment. This means
selecting technologies that are affordable, easy to use, and accessible to the
target audience.
The need for effective communication and dissemination strategies:
ICT projects for agricultural extension should have effective communication
and dissemination strategies in place to ensure that the information and
services being provided reach the target audience. This can include using a
variety of communication channels such as mobile phones, social media, and
radio.

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 The need for monitoring and evaluation: ICT projects for agricultural
extension should be regularly monitored and evaluated to assess their impact
and identify areas for improvement. This can help to ensure that the project is
achieving its objectives and that any necessary adjustments can be made.
The need to integrate ICT with other extension services: ICT projects
for agricultural extension should be integrated with other extension services
such as training, demonstrations, and field visits. This can help to ensure that
the information and services being provided are relevant and useful to farmers.
By taking these lessons into account, organizations can develop more
effective ICT projects for agricultural extension that can help to improve
farmers' livelihoods and increase agricultural productivity.
References
1. Basuroy. T. (2022). Number of active internet users in India 2017-2025.
Statista
2. [Link]
internet-users/
3. Bharat Broad Band Network Limited, Govt. of India, 2023
4. [Link]
5. Deloitte (2022). Technology, Media, and Telecommunications -
Predictions 2022 India Edition
6. [Link]
telecommunications/articles/[Link]
7. Dept. of Science and technology, DST-TARA, Aqua (almost all question
answered) (ICT Based Technologies)
8. [Link]
&Title=Aqua%20(%20almost%20all%20question%20answered%20)&
Category=ICT%20Based%20Technologies&NGOId=1&NGO=Vigyan
%20Ashram
9. Raj. S. (2007). Application of ICTs for Agricultural Extension Services
Provision and its Impact among the Tribal Farmers of Arunachal Pradesh
State of North-East India (e-Arik).
10. [Link]
11. Sun. S. (2022). Smartphone users in India 2010-2040. Statista.
12. [Link]
users-in-india/

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 Chapter - 14
Nuclear Polyhedral Virus (NPV): Role and
Importance for Crop Pest Management

Authors
Mary Lisha J
Ph.D. Scholar, Department of Agricultural Entomology,
Agricultural College and Research Institute (AC & RI), Tamil
Nadu Agricultural University (TNAU), Madurai, Tamil Nadu,
India
Saai Vignesh B
Ph.D. Scholar, Department of Agricultural Entomology,
Agricultural College and Research Institute (AC & RI), Tamil
Nadu Agricultural University (TNAU), Madurai, Tamil Nadu,
India
Balaji K
Ph.D. Scholar, Department of Agricultural Entomology,
Agricultural College and Research Institute (AC & RI), Tamil
Nadu Agricultural University (TNAU), Madurai, Tamil Nadu,
India

Page | 215
Page | 216
 Chapter - 14
Nuclear Polyhedral Virus (NPV): Role and Importance for
Crop Pest Management
Mary Lisha J, Saai Vignesh B and Balaji K

Introduction
Insect-killing viruses, known as entomopathogenic viruses (EPV).
Entomopathogens includes fungi, viruses, protozoa, and bacteria which are
the regulatory operators of pest infestations and important control elements
for insect populations that occur naturally (Roy and Cottrell 2008). The word
entomopathogen was coined by Tanzini et al. (2001) to describe
microorganisms that control the pest population to levels that prevent from
economic harm to crop plants.
Three insect-specifc viruses (Families: Baculoviridae, Polydnaviridae,
and Ascoviridae) are highly host-specifc and non-pathogenic to beneficial and
non-target insects. The baculoviridae has long been regarded as an alternative
to chemical insecticides. The baculovirus (ds DNA) has the best chance of
controlling lepidopteran pests on crops, and it is divided into two categories,
such as: Granulovirus (GV) and Nucleopolyhedrovirus (NPV) (Deka et al.
2021).
The baculoviruses belongs to the family Baculoviridae which is
subdivided into four genera: Alphabaculovirus, Betabaculovirus,
Deltabaculovirus, and Gammabaculovirus (Herniou and Jehle, 2007). Most
important baculoviruses in the Alphabaculovirus genus are: Autographa
californica multiple nucleopolyhedrovirus (AcMNPV), Bombyx mori
nucleopolyhedrovirus (BmNPV), Orgyia pseudotsugata multiple
nucleopolyhedrovirus (OpMNPV), Lymantria dispar multiple
nucleopolyhedrovirus (LdMNPV), Spodoptera exigua multiple
nucleopolyhedrovirus (SeMNPV), and Helicoverpa armigera
nucleopolyhedrovirus (HearNPV) (Blissard and David, 2018).
Nuclear polyhedrosis virus (NPV) was first introduced during 1892 by
Germany into the population of Lymantria monacha in pine vegetation. First
field application of NPV was made by a Californian farmer against the gypsy

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moth (Lymantria dispar), by collecting the NPV infected larva of alfalfa
butterfly (C. eurytheme) in 1913 at USA (Arora et al. 2002 and Moscardi,
1999). As the baculoviruses plays a major role in controlling insect pest and
as alternate for chemical insecticides, whereas it has no residual effect and
shows harmless effect on natural enemies. More than 65 crops like wheat,
maize, sorghum, solanaceous crop, etc., were prone to attack of Helicoverpa
and Heliothis species (Prasun, 2021).
The arthropod specific Nuclear polyhedrosis virus is an obligate pathogen
and it has rod-shaped nucleocapsid which contains double stranded circular
DNA. Baculovirus involves in the production of two virion phenotypes which
carries as identical genome such as occlusion-derived virus (ODV) and
budded virus (BV). Nucleocapsids are enclosed within an envelope called
virions consisting of a proteinaceous polyhedral occlusion body inside which
the virions or virus rods are embedded (Grula et al., 1981 and Beniya et al.,
1996). In ODV-virions, the nucleocapsids within the nucleus and virions
acquire an envelope, whereas in Occlusion body (OB), the virions are
enveloped within a crystallized protein matrix. After ODV infection on midgut
cells, the BV virions bud from midgut epithelial cells and onto hemocoel then
transmit systemic infection from cell to tissue of whole body of insect (nuclei
of cell tissue, fat bodies, haemocytes, tracheal matrix, ganglia and brain) due
to alkaline gut juice which liberated the viruses (Harrap, 1972 and Milks et
al., 2003).
Mode of action/infection
Primarily the infection occurs by ingestion of virions or polyhedra. When
the susceptible larva consumes the food with the polyhedral, the occlusion
proteinaceous bodies were dissolved under the alkaline condition of gut with
pH of above nine (Nawaz et al., 2019). The midgut epithelial cells get infected
by virions, later the nucleus of infected cells will produce the new virions and
infect the cells of haemocoel and other tissue (fat body). Until the cell lyse,
the occlusion of virions or polyhedral takes place in that tissue. Few days later,
the larva die and spilled on soil and foliage, the polyhedron might disperse to
other insect by ecological factors (Entwistle et al., 1983; Rahman and
Gopinathan, 2004).

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Symptoms of NPV infected pest
After infection, insect larva become inactive and dull colour which in turn
reddish pink on ventral surface. While peak infestation of NPV, the larva
become flaccid and fragile. The unique symptom of infected larva hang upside
down on foliage by silken thread, this phenomenon is called “wipfelkrankheit”
or “tree top disease” (Babu et al., 2009).

Mass production of NPV of Spodoptera litura

The early fifth instar larval stage of Spodoptera litura was chosen for
mass production of NPV, which yields maximum amount of NPV. Host
culture (S. litura) should be maintained throughout the mass production
(Brown and Dewhurst, 1975; Takatsuka et al., 2007).

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 By using the castor leaves/natural diet under laboratory condition the S.
litura can be mass cultured in plastic buckets

For overnight pre-starve the fourth instar larvae

Virus suspension containing 1 x 108 POB/ml in water containing 0.1% teepol

(sticking agent) was prepared

Castor leaves was dipped in viral suspension and shade dried

Larva was forced to feed on treated castor leaves for two days

Then diseased larva was collected in distilled water and allowed to putrefy

After 5 days, the polyhedral settles at bottom (white layer)

Sediment contains POB, which is suspended in distilled water and

centrifuged at 500 rpm for one minute

The supernatant contains the POB was again centrifuged at 2500 rpm for 15
minutes and pellet was collected

Repeat the differential centrifugation for obtaining pure POB’s

This method was similar to the mass production of Helicoverpa armigera
NPV. The dose of virus (Pure POB’s) is expressed as larval equivalent (LE),
(1 Larval Equivalent = 6 x 109 POB’s).
In-vivo NPV production
The modern method of NPV mass production was carried out in France,
Canada and USA. It involved the computer-controlled robotics operated
techniques for mass production with capacity of 0.25 to 1 million hectare per
annum (Tinsley and Kelly, 1985; Lindegren et al., 1993). Steps involved in
in-vivo mass production are:
1) Infection of NPV on susceptible larva

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 2) Infected larva was grown for specific time to increase the infection
rate thereby the virus gets proliferate
3) Finally, the infected larva was collected and virus get extracted
Major insect pest and their management
Spodoptera litura NPV (SlNPV) and Helicoverpa armigera NPV (HaNPV)
It is a polyphagous pest which meets the global importance due to its
insecticide resistance and damaging range. Application of SlNPV as alternate
for chemical insecticide shows good control of Spodoptera litura on cole crops
(Takatsuka et al., 2007; Kumari and Singh, 2009).
Helicoverpa armigera is the most important lepidopteran pest in South
Africa and serious pest of citrus, where they laid eggs on or near the
inflorescence during spring season (Coffee, 1980; Arora and Padmanaban,
2002).
Mode of application of SlNPV
Filed application of two sprays at 500 litre/ha, first and second spray @
45th day and 60th day after transplanting respectively.
Mode of application of HaNPV
Reduction in H. armigera larval infestation within 7days by application
of POB at concentration of 1.15 x 107 OBs/ml of HaNPV on tomato foliage.
List of pests associated with crops that managed by NPV
Pest
Crop NPV References
Common name Scientific name
(Hackett et al., 2000 and
Corn earworm Helicoverpa zea Maize HzNPV
Rowley et al., 2011)
Tobacco Heliothis (Ignoffo and Couch,
Cotton HvNPV
budworm virescens 1981)
Cotton,
Cotton Helicoverpa (Moore et al., 2004 and
Tomato, HaNPV
bollworm armigera Nawaz et al., 2019)
Wheat
(Kumari and Singh,
Vegetables,
Tobacco Spodoptera 2009; Takatsuka et al.,
peanuts and SlNPV
cutworm lituura 2007; Tinsley and Kelly,
rice
1985)
Spodoptera (Hussain et al., 2002 and
Army worm Maize SeNPV
exigua Murray et al., 1995)
Spodoptera
Fall army worm Maize SfNPV (Podgwaite et al., 1979)
frugiperda

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 Autographa Cabbage,
Alfalfa looper AcNPV (Sun and Peng, 2007)
californica cotton
Lymantria
Gypsy moth Forest crops LdNPV (Kool et al., 1991)
dispar

Constrains in production of NPV

• Cross contamination of culture by other pathogen that reduces the
yield and quality of the product (NPV).
• Non-availability of periodical supply of healthy larvae.
• Reduced production rate per insect over time.
• Some of the important factors those affect the mass production are:
• Poor quality insects
• Inappropriate dosing
• impure inoculum
• Poor rearing
• Unsuitable harvesting
• Poor sanitation
Effective application of NPV
The effectiveness of NPV in field was enhanced by:
• Spray during early morning or late evening to safeguard the microbes
from sunrays.
• By adding 1 ml of robin blue (UV light absorbent) to a litre of spray
solution was improved the efficiency of NPV.
• By adding the phagostimulant (jaggery) and adhesive (teepol) helps
to improve efficacy.
• The dosage of about 250-500 LE/ha was recommended depends upon
the foliage density.
Application systems
High-volume sprayer
High-volume applications were made with an azo-propen hand sprayer
using a birchmeier helicon sapphire nozzle100, pressure 4 bar. This sprayer
nozzle should be 50 cm over the crop during spray time (Bateman et al., 2007).

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Low-volume sprayer
Low-volume spinning disc applications were made by micron-ulva eight
spinning discs. This sprayer nozzle should be 75 cm over the crop during spray
and droplets angel should be 50 degrees. Volume should be applied 3 ml of
virus suspension in water with 20% mineral oil per meter square (Bateman et
al., 2007).
Ultra-low volume
In ultra-low volume electrodynamic sprayer, use of electrod provided by
ICI. In this, the sprayer nozzle position should be 50 cm above the crop
(Bateman et al., 2007).
Conclusion
NPV is a bio pesticide can be an effective alternate to conventional
pesticides as they are renewable sources with reduced risk to both human and
the environment. Hence NPV can making it ideally suited to include in an
integrated pest management approach.
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Interactions between coccinellids and insect-parasitic fungi. European
Journal of Entomology, 105(3).
27. Takatsuka, J., Okuno, S., Ishii, T., Nakai, M., & Kunimi, Y. (2007).
Productivity and quality of polyhedral occlusion bodies of a
nucleopolyhedrovirus harvested from Spodoptera litura (Lepidoptera:
Noctuidae) larvae. Applied entomology and zoology, 42(1), 21-26.

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28. Tanzini, M.R., Alves, S.B., Setten, A., & Augusto, N.T. (2001).
Compatibilidad de agentes tensoactivos con Beauveria bassiana y
Metarhizium anisopliae.
29. Tinsley, T.W., & Kelly, D.C. (1985). Taxonomy and nomenclature of
insect pathogenic viruses.

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 Chapter - 15
Canopy Management

Authors
Megha Patidar
Sher-e-Kashmir University of Agricultural Sciences and
Technology, Jammu, Jammu and Kashmir, India
Parshant Bakshi
Sher-e-Kashmir University of Agricultural Sciences and
Technology, Jammu, Jammu and Kashmir, India

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Page | 228
 Chapter - 15
Canopy Management
Megha Patidar and Parshant Bakshi

Introduction
Canopy: The physical aspects of a tree, including the stem, the branches,
the shoots, and the leaves.
Canopy management: The establishment and maintenance of tree
structure in relation to size and shape for the highest yield, productivity, and
quality
Canopy architecture management: A set of operations on the plant's
surface with the goal of maximising the output of high-quality fruit per square
foot.
Particularly since technological advancements were used, too vigorous
trees have become commonplace. This is especially true for plantings with a
high density, where the advantages of better soil preparation, irrigation,
nutrition, fertilization, pest and disease control, and weed control measures
have led to higher tree vigour and, typically, increased within-canopy shadow.
Techniques to avoid shade have been developed through recent study on
canopy management.
Enhancing the amount of dry matter partitioning towards reproductive
portions by management of canopy design is a key strategy for enhancing crop
performance. Management of canopy architecture generally refers to the
positioning and upkeep of plant framework in connection to optimum fruit
production. The development of a strong primary branch framework, wider
crotches in scaffold branches, healthy and evenly distributed secondary and
tertiary branches, enough fruiting terminals in the most productive areas,
healthy foliage with high photosynthetic efficiency to maximise the efficiency
of solar radiation use, and enough space for air circulation within the canopy
are all components of appropriate canopy management.
The fundamental idea behind three-dimensional approaches to canopy
management of a perennial tree to maximise the use of the land and climate
parameters for higher yield. One of the most crucial management techniques

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for fruit plants is canopy control. This form is based on the precociousness and
early longevity of the fruit trees in the orchards.
The quantity and quality of fruits are improved by a number of processes.
One of these is canopy management, which is crucial for maintaining fruit
supply and quality. By controlling the tree canopy, fruit quality can be
improved.
Features of an ideal canopy
• Allow enough ventilation and light
• Maintain sufficient foliage and shield the fruits from sunburn
• Avoid having foliage overlap.
• Dwarf, open, and spreading
• A sufficient number of fruiting units
Principles of canopy management
• Maximum use of available light.
• Avoiding the development of a microclimate that is conducive to
disease and pest infection
• Convenience in doing cultural activities.
• Improving yield, productivity, and quality to the fullest.
• Cost-effectiveness in acquiring the necessary canopy architecture
• Rejuvenation of huge, overgrown orchards with deteriorating
production and fruit quality.
• Small trees are more effective than huge trees at capturing sunlight
and turning it into fruit.
• A decrease in the extra cost associated with harvesting huge trees.
• A safety risk exists when larger trees are harvested.
The amount of sunlight that trees are able to intercept depends on the
canopy management, especially on its elements like tree training and pruning,
as the shape of the tree affects how exposed its leaf area is to incoming
radiation. An optimal training approach concentrates on the arrangement of
plant components, in particular, to produce a superior plant architecture that
maximises the use of sunlight and encourages productivity.
Trees and their fruits require light for proper growth and development.
The green leaves capture the sun's energy to create sugars and carbohydrates
which are then transferred to the locations where they are required, such as

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buds, flowers, and fruits. The amount of light that is accessible and is
intercepted by the tree canopy but does not reach the orchard floor is referred
to as light interception. The ability to intercept is influenced by plant density,
branch and tree direction, canopy size and form, and leaf area index within the
canopy. The horizontal rather than the vertical component of the tree proved
more important for interception. The growth, productivity, yield, and fruit
quality of trees are all improved by increased light penetration into the tree
canopy. Light penetration in an orchard is also influenced by planting density
and orientation. In general, rapid shadowing becomes an issue with close
planting. Compared to tree orientations towards the west and south, an east-
west row produces more shading. Larger fruits typically develop on strong
bearing branches. The challenge for a fruit farmer is to first establish a sturdy
and balanced framework for the trees before outfitting them with the proper
fruits.
Why we need canopy management?
The growth, productivity, yield, and fruit quality of trees are all improved
by increased light penetration into the tree canopy. Light penetration in an
orchard is also influenced by planting density and orientation. In general, rapid
shadowing becomes an issue with close planting. Larger fruits typically
develop on strong bearing branches. The challenge for a fruit producer is to
first establish a solid and balanced architecture of trees before providing them
with the proper fruiting. It goes without saying that pruning done in the early
years must be of the training variety to produce a sturdy structure with limbs
spaced evenly or any other desired shape.
The main objectives of canopy management are given below
1) To obtain a better yield of high-quality goods.
2) To keep the growth of the roots and the shoots in proper balance.
3) The development of strong crotches or a crotch angle.
4) To eliminate extra, crowded, dead, diseased, and pest-affected
shoots.
5) To control tree architecture or create the desired shape for a planting
system with a high density.
6) To make management procedures like spraying, harvesting, etc.
easier.
7) To effectively utilise temperature, light, and air.
8) To control how much light and air a plant is exposed to.
9) To allow for equipment access between rows

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Importance of canopy management
1) Producing more fruit from smaller trees has increased productivity
and fruit quality in several fruit crops.
2) Rejuvenation of large, overgrown orchards with falling production
and fruit quality.
3) Compared to giant trees, little trees are better at capturing sunlight
and turning it into fruit.
4) A decrease in the extra cost associated with harvesting huge trees.
5) The harvesting of larger trees poses a safety concern to the workers.

Type of canopy management

6) Columnar type- Trees are tall, slender, and upright branches. Their
vertical form gives them height and serves as screening. Malus spp.
and Prunus persica are two examples.
7) Pyramidal and conical shape type- Pyramidal tree with horizontal
branches and a main central trunk that is wider at the bottom.
Depending on the height of the trunk, the branches may begin at
ground level or higher. Similar to a conical form, but slenderer and
more shaped like a bullet. The full growth of this tree requires space.
Christmas tree, as an example.

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 8) Vase type- An upright, bending shape with a widest point at the top,
branching from a central trunk. Vase-shaped canopies are elegant and
ideal for walkways because they provide shade.
9) Round or oval type- The centrals are erect, with a strong trunk that
branches into a dense crown that is both round and oval in shape.
They provide substantial shade and may grow so much foliage that
the branches are hidden.
10) Spreading or open shaped type- This form of canopy has an open,
asymmetrical design that may be wider, like cherry, mango, and
sapota, among other fruits.
11) Weeping shaped canopy- Branches in this canopy droop and sweep
the ground. Examples include bottlebrush, weeping willow, and
weeping cherry.
Fruit crops have a seasonal and a lifetime development pattern for canopy
management. Canopy alteration has both long-term benefits and drawbacks.
Fruit trees with an unmanaged canopy provide many challenges, including
difficult spraying, pruning, and manual harvesting, as well as inadequate light
penetration and interception across the canopy. Low leaf area index, early light
interception, and the percentage of land covered by canopy all contribute to
delayed and subpar cropping. Due to these drawbacks, there has been a broad
attempt to alter the perennial fruit tree canopy model by increasing tree
density, reducing tree size, and using other horticultural methods to improve
light dispersion.
Techniques for canopy management
Pruning
Pruning is described as the removal of plant parts to create a desired
canopy architecture and to diminishes the foliage density by cutting off the
branches of trees that aren't producing fruit. The primary goal of tree pruning
is to create a balance between vegetative and reproductive growth, which will
result in an annual harvest of high-quality fruits. Pruning can be done to
enhance tree shape, control a tree's development, blossoming, and fruitfulness,
improve the quality of the fruit, fix damage, contain a plant, and promote light
and spray penetration.
Types of pruning
Based on time of pruning
Dormant pruning: Similar to summer pruning, however cuttings are
conducted before active new growth has started during the dormant season.

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Light penetration is guaranteed by the architecture of the tree. The removal of
dead, diseased, and dry branches is the least common practise for pruning,
inspective of varied conditions. An organism can enter through a sick branch
or a troublesome sub that cannot heal over. As a result, it becomes a constant
source of infection. Jagged and damaged branches with wounds that cannot
heal over should also be removed, as should those that wear the bark of
adjacent branches. The ideal pruning is one that is carried out once a year to
get the desired physiological effects and to make the smallest cut size possible.
Additionally, during the growing season, when water sprouts appear and
unhealthy branches and those that could harm nearby plants should be cut as
soon as possible, this should be complemented with some little care.
Summer pruning: Most fruit species not only exhibit dormancy but also
enter a rest period in the summer when their shoots stop developing but their
leaves continue to produce photosynthates until leaf fall in the autumn. during
the growing season, the selective removal of shoots or branches. By cutting
off the vigorously growing branches, cropping zone light intensity and colour
intensity were boosted.
Early summer pruning: After the flowering season, when the shoot
growth is still succulent, early summer pruning is done. The buds directly
behind the spots where cuts are done are encouraged to begin growing new
shoots. The sprouts still on the tree may gain vigour as a result.
Mid summer pruning: A tree has completed the majority of its annual
shoot growth, and almost all of its bud development has taken place. At this
time, pruning may result in a few shoots emerging from wounds; these shoots
will also gradually expand. The next spring, though, is when they reach their
maximum size.
Late summer pruning: A tree will not grow until the next spring if it is
pruned after midsummer, when it is fully dormant. Since the leaf surface is
decreased, it also has a negative impact on root growth. However, it lasts for
a shorter time than a tree pruned in the middle of the summer.
Based on the severity
Heading back: The level terminal portion of the shoot is eliminated; for
example, one-fourth or two-thirds of the shoots may be headed back; this
pruning encourages the formation of secondary branches.
Tipping or pinching: A flower bud will sometimes grow at the bud that
is immediately below the cut made when a shoot is trimmed or pinched back
in the summer. This only occurs when the wood is in a specific vegetative

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condition, which can only be understood by experience. the act of removing
the apical bud from shoots or spurs while they are still growing. The highest
bud on the stalk, in especially the leaves close to it, experiences a rapid but
brief buildup of carbohydrates and hormones that produce flower buds when
it is pinched or lightly trimmed back.
Notching: It was used as a training tool to activate dormant buds before
to the onset of rapid growth. On canes that are around 8 months old, 1-2 buds
are often chosen for notching in the central section. For example, Poona fig.
Notching involves removing of small slice of bark immediately above the
dormant bud, giving 2 slanting cuts as deep as the bark. Notch should be 2.5
cm long and the breadth depends on thickness of the shoot.
Dehorning: It consists of removal of all the wood after leaving 7-10cm
thick stub all over the tree. Such severe treatment should normally not be given
except when it is the last resort to bring it into commercial bearing.
Thinning out: It is the complete elimination of a shoot or branch at the
site of insertion on a big branch or scaffold. Due to their opening effect, these
wounds allow sunlight to pass through and reach the tree's branches.
Training
Training is the most advanced type of pruning, usually done in the first
few years after planting, with the goal of shaping the plant and creating an
organizational behaviour of scaffold branches that can support a heavy fruit
load without limb breakage. To do this, plant portions may be pruned with or
without the use of trellising, spreading, tying, or pergolas.
The main goals of training are to reduce vegetative vigour, improve fruit
and foliage exposure to light, boost airflow to reduce the spread of disease,
and simplify mechanised trimming and harvesting techniques. Utilizing both
vertical and horizontal space should be taken into consideration while
deploying the training systems. The selection of a training system influences
planting distances, light interception, and ultimately the performance of the
orchard in terms of the yield of fruit in terms of both quality and quantity. In
many orchards, the following training techniques are employed to regulate tree
growth:
Delayed open centre: The tree is first trained to become a leader type by
leaving the centre axil unpruned for the first four or five years. Then, like in
an open centre system, the core stem is headed back and lateral branches are
permitted to spread out. Fruiting trees should be pruned to prevent
overpopulation in the orchard, while immature plants must be trained to

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develop a solid foundation and avoid weak crotches. The plant needs to be
trained to grow as low-headed trees so that it can be picked by hand repeatedly.
Central leader or modified central leader system: In this strategy, the
centre leader branches are permitted to spread out on their own so that they
can expand more quickly and aggressively than the side branches, which will
result in a taller tree. Near the top of the tree, the fruit production is greater.
Less strong and prolific are the lower branches. In favour of a central trunk
with side branches, weak side shoots are pruned back.

Source: Gelley, C., 2018. Pruning fruit tree

Spindle bush system: This system was first developed in Germany. It is
either a modified dwarf pyramid or a transitional form between a vertical
cordon and a bush form. It differs from the dwarf pyramid in that it lacks a
distinct scaffold branch arrangement, and it differs from the vertical cordon in
that the fruit is produced on short branches as opposed to the main stem or
trunk. The most crucial aspect of this technique is the horizontal placement of
lateral shoots with little to no summer pruning. With a straight central leader
and several little fruiting branches, it is trained with or without support posts.
Spreaders twist these branches out and downward to create wide crotches and
encourage early fruiting. Tree spread is controlled by cutting back the shoots
to ½ to ¾ of their length or back to weak laterals.
Overhead trellis or Bower system of training: It creates a pleasant
microclimate within the vine canopy and minimizes the negative effects of dry
and hot weather on the metabolism and longevity of the vine. In this technique,
vines are strung over a pandal that is suspended between 2.2 and 2.4 metres
above the ground on pillars with holes spaced 60 cm apart. The vines are either
stacked or tied to each other to prevent them from growing over the height of
the wires.
Cordon system: This system was created by Breuil in France in the 19th
century. Single stem trees are called cordons. Depending on the angle at which
the stem is trained, cordon might be vertical, oblique, or horizontal. Peaches
have been taught to line up against a wall.

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 Kniffin system: This method uses two vertically supported trellises of
wires. Four canes, one along each wire, and bearing branches swing freely
thanks to the vine's training. Two wires are horizontally extended in this
arrangement at heights of 0.90 and 0.60 metres. It is advised to place vines 2-
4 metres apart between two poles. As a result, it's crucial to adopt the right
system according to the crop as yields can vary.
Espalier: The approach is comparable to the kniffin system of grape
training. The trellis is 5 feet tall and has either 2 or 4 wires spaced 3 and 5 feet
apart.
Palmette: The shape of a palm leaf, an open fan, or an open hand with
spread fingers is referred to as a palmette. There are several types of palmettes,
including candelabera, fan, and horizontal.
Tatura Trellis system: The irrigation research institution Tatura in
Australia developed one of the highest crop yielding systems. Each hactare
can hold 1600-2000 trees. A Y-shaped leader that is connected together with
wires makes up the primary framework.

Bending: Physically bending an apple tree limb causes auxin, a growth

hormone, to be redistributed and the growth of terminal shoots to be reduced.
It has been demonstrated that bending stresses increase the amount of ethylene
in the branch's interior air spaces. Because bent limbs have less vegetative
vigour, they produce less gibberellins, which are harmful to flowering.

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Additional locations for flower production are produced by the development
of fairly robust lateral stems and spurs. Early fruit development and more
flowering are the results of these two actions. Improved light penetration and
the fact that more fruit hangs freely rather than rubbing against the branch on
more upright limbs both contribute to an improvement in fruit quality.
Scoring, girdling or ringing: It has been demonstrated that scoring and
ringing, which stop the phloem's downward flow of hormones and
carbohydrates, change growth and fruiting. The least impact is from scoring,
which is a constraining incision through the bark but not into the wood.
Ringing, which involves removing varying amounts of bark in a
circumscribing ring, gets worse as more bark is removed until the tree dies.
Use of growth regulators/retardants
Growth retardants are a class of synthetic chemicals that act as
bioregulators, altering a plant's growth and developmental behaviour without
causing phytotoxicity or malformation. When used in the right concentration,
these compounds have typical effects on plant architecture, such as
a. Inhibition of shoot growth (plant height, internode elongation, leaf
area), while maintaining the same number of internodes and leaves
and intensifying the green pigmentation of the leaves
b. Maintenance or slight promotion of root growth (main roots often
longer and thicker).
The root-to-shoot ratio is altered in both situations in favour of the root.
Chemicals can be used to control plant height in at least three different ways:
• By severly suppressing apical meristematic activity or by destroying
the terminal buds or branches.
• Through preventing interned elongation while maintaining apical
meristematic integrity
• Reducing of apical control.
With three sprays of maleic hydrazide, a significant reduction in shoot
length was seen (MH 500 ppm at leaf stage followed by 1000 ppm at leaf stage
and 1500ppm at 15 leaf stage) In the Thompson Seedless grape, maleic
hydrazide (MH) appeared to be more effective than CCC at boosting cane
diameter (Shikhamanyand Reddy, 1989). It was discovered that applying
cultar (25% paclobutrazol) dramatically reduced the growth of annual shoots
and enhanced photosynthetic activity, both of which may have increased
cherry output. Paclobutrazol 10 g/tree applied to mango trees resulted in

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decreased tree volume (33.1%), height (21.20%), and mean shoot length
(48.2%). This response was related to paclobutrazol's GA-inhibitory activity
(Murti et al., 2001) It was discovered that adding 1500 ppm CCC to red
raspberries of the cv. "Autumn" improved the number of fruiting buds (Ghora
et al., 2000).
Conclusion
Unmanaged tree canopy not only lowers fruit production but also lowers
the quality of the harvest. Higher productivity is made possible by it, as are
sustainable cropping, high, consistent yields, and enhanced farm management
techniques. This management strategy offers high and consistent yields of fruit
of high quality while requiring little labour to keep up with rising production
expenses. Canopy management increases output, boosts fruit quality, supports
cultural traditions, and aids in pest and disease management. In new
plantations, early training and pruning are given to establish strong tree
frameworks, however in older plantations, the goal of canopy management is
to reduce tree height and provide for solar radiation inside the canopy by
thinning out excessive biomass.
References
1. Balamohan Thangappan, Balraj Gopu, R, Balakumbahan and R, Anitha.
(2022). Canopy Management A perspective studies in India and Abroad.
In book: ICAR sponsored winter school on Multi-stored cropping system
and Canopy Architecture Management in Horticulture crops. Delhi-
110001.
2. Gelley, Christine. (2018). Pruning Fruit Tree. Noble Country Agriculture
and Natural Resources.
3. Ghora, Y., Vasilakakis, M., Stavroulakis, G. Effect of growth retardants
(CCC, daminozide & Paclobutrazol) ongrowth and development of red
raspberries cv. ‘Autumn Bliss’ cultivated under plastic green house
condition in China- crete, Greece. Acta Horticulturae, 513: 453-458.
4. Kumar, K. and Sharma, S. (2020). Canopy management in fruit crops.
Just Agriculture, 1(4): 209-215.
5. Murti, G.S.R., Upreti, K.K., Kurian, R.M. and Reddy, Y.T.N. (2001).
Paclobutrazol modifies tree vigour and flowering inmango cv.
‘Alphonso’. Indian Journal Plant Physiology, 6 (4): 355-360.
6. Practical Manual on Canopy Management in Fruit Crops- Dr. Gorakh
Singh (2010). Department of Agriculture & Cooperation, Ministry of
Agriculture, Govt. of India, New Delhi.

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7. Rajkumar, Sapna and Sharma, K.D. (2021). Canopy management in fruit
crops. AGRI MEET. 2(01): 1-7.
8. Shikamany, S.D. and Reddy, N.N. (1989). Effects of growth retardantson
growth, yield and quality in grape cv. Thompson seedless. Indian Journal
of Horticulture, 46(1):31-38.
9. Vandana, Solanki, S.P.S. and Lamo, K. (2017). Canopy management:
Way to develop fruit tree architecture. Biomolecule report. ISSN No.
2456-8759.
10. Wani, R.A., Din, S., Khan, M., Hakeem, S.A., Jahan, N., Lone, R.A. and
Khan, J.A. (2021). Canopy management in fruit crops for maximizing
productivity. International Journal of Chemical Studies, 9(3): 160-165.

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 Chapter - 16
Breeding for Improved Quality in Vegetable
Crops

Authors
Sheetal Rana
XXX

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 Chapter - 16
Breeding for Improved Quality in Vegetable Crops
Sheetal Rana

Abstract
Vegetables play a very important role in human diet by supplying
important natural elements in a balanced manner which are generally deficient
in other food materials except fruits. They contribute functionally active
secondary metabolites that protect against a variety of health problems,
particularly non-communicable diseases and are excellent sources of
roughages, proteins, vitamins, carbohydrates and minerals, which are usually
required for perfect health. They are cheaper sources of nutrients and can meet
the dietary requirements very easily. As the demand for vegetable crops is
continuously increasing globally mainly due to increased health awareness
and changes in eating patterns, there is more emphasis on improving the
quality and value addition of vegetable crops. For this, various techniques and
methods have been developed, but these traditional breeding methods have
been slow and inadequate, so new developments in molecular and biochemical
diagnostic tools and techniques have shown promise in improving
nutraceutical content and other quality traits. Molecular markers, next-
generation sequencing, RNA interference (RNAi) and genetic engineering
plays an important role in reducing the time and cost associated with quality
breeding.
Introduction
Vegetables are essential for a well-balanced diet because they provide
vitamins, minerals, dietary fibre, and phytochemicals. Vegetables play an
important role in Indian agriculture in terms of food and nutritional security.
They are commonly referred to as "protective food" due to their ability to
protect against various kind of diseases. Vegetables are also regarded as the
century's nutraceutical food. Almost every vegetable contains bioactive
compounds such as vitamins, anthocynins, flavanoids, carotenoids, and
polyphenols. Because of their antioxidant properties, all of these compounds
are capable of disease prevention and reduction. Until recently, vegetable crop
improvement was largely limited to conventional breeding approaches, which

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rely on interspecific sexual hybridization of plants with desirable heritable
characteristics and naturally or artificially induced random mutations.
Improvements can be obtained through reassorting, which has been
accomplished by improving breeding technologies through randomly induced
changes and by creating completely new possibilities through biochemical
engineering. As a result, combining traditional breeding methods with
biotechnological tools will be promising for crop improvement strategy in the
future. There is need to develop new methods and technologies
for advancement of vegetable through biotechnological tools to provide food
and nutritional security.
Nutraceuticals in vegetables
Vegetables are an important part of a balanced diet because they supply a
significant portion of the dietary demand for minerals and vitamins.
Nutraceuticals are important components of quality attributes in vegetable
crops. Main emphasis of consumers is the vegetables rich in carotenoids,
anthocyanins, flavonoids, glucosinolates, capsaicin, oleoresins and terpenoids
due to their potential health benefits. Nutraceuticals and various bioactive
compounds present in vegetable crops like Allyl sulfides in Allium vegetables
(garlic, onions, chives, leeks) protect against certain cancers and heart
diseases, boost the immune system; glucosinolates from Cole vegetables play
an important role in protection against cancer, heart disease and stroke. β-
carotene is important for healthy skin and membranes, immune system and
eye health; lycopene from tomato, watermelon, carrot and red peppers useful
against prostate cancer; ascorbic acid rich green peppers, broccoli, green leafy
vegetables, cabbage and tomatoes is helpful in growth, development and
repair of all body tissues and absorption of iron from foods. The momordicin
and charantin from bitter gourd have anti-diabetic properties and help in blood
purification (Behera and Singh, 2019). India is endowed with an abundance
of wild and cultivated vegetable crops which are high in antioxidants and
dietary minerals. There is a lot of variety within vegetable species such as
antioxidants and lycopene in tomato, capsaicin in chilli, lycopene in
watermelon and glucosinolates in cauliflower and broccoli (Kushad et al.
1999).
Quality trait- Quality generally refers to various aspects like colour, size,
nutrient content, shelf life and suitability for processing and a trait defines
aspects of quality of produce. Each crop has specific and different set of
quality traits but they differ from species to species depending upon plant part
used as economic product.

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Quality attributes of vegetables
1. Intrinsic quality attributes
• Sensory attributes
• Health attributes
2. Extrinsic quality attributes

Breeding objectives for quality vegetables

Vegetables are perishable foods, and obtaining an adequate supply on a
consistent basis remains a major challenge. As a result, measures to increase
vegetable production through home gardens, kitchen gardens, container
gardening, roof gardening and other methods attracts not only people living in
the rural areas but also people living in urban areas. This requires
identification of region-specific vegetable crops and breeding of varieties high
in dietary nutrients with a longer harvest period, adaptability to growing
conditions and widespread acceptance among the ultimate beneficiaries.
Breeding goals could be presence of high levels of dietary nutrients and
antioxidants, good organoleptic scores, tolerance to partial shade, a high
proportion of edible fruits/parts, a short gestation period, resistance to diseases
and pests, responsiveness to organic sources and so on. Generally various

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techniques and methods have been developed to improved varieties which are
grown commercially, but more efforts are required to develop plant types
which are ideal for home gardens or other micro-scale production systems. In
general, vegetable breeding for nutrients is focused on high yield and dietary
micronutrient richness, which are traditional traits but enrichment of
nutraceuticals requires special attention from identification of suitable
germplasm to proper handling of the breeding programme and maintainance
of improved varieties. Consumer preferences change over time and it affects
the futuristic traits. These traits vary from crop to crop like for cole crops,
presence of β-carotene, glucosinolate, anthocyanin, iron and zinc, their
stability in different growing temperature regimes, better retention of
bioactive compounds during post-harvest life and mild flavour, in case of
tomato there is need to develop varieties rich in minerals and vitamins, having
high lycopene content which is stable at high temperature, TSS and strong
antioxidant activity. The demands for biofortified vegetables is increasing day
by day so there is a need to develop nutrient enriched varieties at a faster rate
to meet the market demands. For the success of a quality breeding programme,
a combination of robust genetic sources, modern genomics tools and
biochemical diagnostic techniques are required. For breeding nutrient rich
varieties, the pre-requisites are:
a) High nutrient density must be combined with high yields and high
profitability
b) When consuming the biofortified varieties, the micronutrient status
of human subjects must be shown to improve
c) While processing and cooking, sufficient nutrients must be retained
and must be sufficiently bioavailable
d) Farmers must adopt the enriched/biofortified varieties and ensures
more availability to those suffering from micronutrient malnutrition
in significant numbers
Breeding methods for quality traits
Nutrient enhancement in vegetable crops is accomplished by combining
one or more micronutrients from germplasm using traditional and/or advanced
breeding methods. The use of molecular markers for the identification and
transfer of gene(s) and QTLs for nutrients, as well as their use in
biofortification, is becoming more common in vegetable crops. The success
of breeding program mainly depends on preciseness of genetic architecture
and effective breeding system. Nature of gene action and relative magnitude
of additive genetic variance, dominance variance and epistasis present in the

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breeding population guides the choice of breeding method while effective
selection and manipulation of favourable gene combination, additive genetic
variance and exploitation of dominance variance is important for efficient
breeding procedure.
Advanced techniques in vegetable breeding
• Mutation breeding
• Polyploidy breeding
• Molecular breeding for nutrient enrichment
• Genome sequencing and transcriptome analysis for quality traits in
vegetables
• Transgenic approach for quality improvement
• RNA interference in vegetables for quality traits
• Targeted genome editing
Mutation breeding
Mutation is a sudden heritable change in the DNA sequence leading to
change in the characteristics of an organism. In vegetable breeding when we
treat a biological material with a mutagen in order to induce mutations is
known as mutagenesis. Different types of mutations are there viz., spontaneous
mutations, induced mutations, genomic mutations, structural mutations, gene
mutations, dominant and recessive mutations.
Various application of mutation breeding are:
1) To induce desirable mutant alleles, which may not be present in the
germplasm
2) In improving specific characteristics of a well-adapted high yielding
variety.
3) To improve various quantitative characters, including yield.
4) In order to increase genetic variability, F1 hybrids developed from
intervarietal crosses may be treated with mutagens and by facilitating
recombination among linked genes.
5) To transfer à chromosome segment carrying a desirable gene from
the alien chromosome to the chromosome of a cultivated species,
Irradiation of distant hybrids has been done.
Various mutant varieties have been developed globally and out of 3000
mutant varieties 776 have been induced for nutritional quality (Jain and

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Suprasanna 2007). In case of tomato, high pigment-1 (hp-1) mutation known
to increase flavonoids content in fruits (Sapir et al. 2008). Spontaneous
mutation in case of cauliflower resulted in ‘orange’ cauliflower which was
found to be very rich in betacarotene (30-100 ppm) than white fleshed (2 ppm)
ones (Li et al. 2001). Another option to identify allelic variants for targeting
genes in natural collections is Targeting Induced Local Lesions in Genomes
(TILLING) and EcoTILLING (Comai et al. 2004). In tomato crop
improvement TILLING technology was used to develop new mutant genetic
resource (Mianoia et al. 2010).
Polyploidy
Polyploidy can be induced due to aberration in cell division. This can
occur both in the mitosis as well as in meiosis. This technique can be used
successfully in vegetable breeding as a means of enhancing nutraceuticals and
colour. Quality of vegetables like radish, pumpkin, muskmelon and
watermelon has been improved using this technology. In a study, it was
observed that soluble solid, soluble sugar and vitamin C contents were rich
and higher in tetraploid muskmelon as compared to the diploid fruit (Zhang et
al. 2010). Liu et al. (2010) observed that the range of lycopene content among
tetraploid watermelon fruits was from 38.1 to 59.8 mg/kg whereas in triploid
watermelon fruits was 41.2 to 61.8 mg/kg and in diploid watermelon fruit was
from 33.2 to 54.8 mg/kg. They stated that lycopene content in the triploid and
tetraploid was higher as compared to diploid. Marzougui et al. (2009)
observed that 0.5% colchicine solution induced polyploidy in Trigonella
foenum-graecum L. and that the autotetraploid cultivar posessed larger leaf
area and larger productivity compared to the diploids and the leaves were
found to be rich in sodium, potassium, calcium and phosphorus.
Molecular breeding for nutrient enrichment
Molecular markers enable clear identification of lines/individuals in a
segregating population for handling of quality traits that is otherwise difficult
to phenotype due to a lack of technical expertise and availability
of sophisticated equipment. In general, quality traits are the result of various
complex pathways and are governed by quantitative traits which are
generally influenced by the environment. Molecular markers such as random
amplified polymorphic DNA (RAPD), microsatellites or simple sequence
repeats (SSR), inter-simple sequence repeats (ISSR), sequence characterized
regions (SCAR), sequence tag sites (STSs), cleaved amplified polymorphic
sequences (CAPS), single nucleotide polymorphisms (SNPs), expressed
sequence tags (ESTs) and diversity arrays technology (DArT) are used to

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study linkage with gene responsible for the presence of high nutraceuticals
and edible colours by using mapping population. A marker is used for indirect
selection of a trait of interest in case of MAS (Marker Assisted Selection). To
identify the molecular marker linked to genes, mapping populations such as
Near Isogenic Lines (NILs) and Recombinant Inbreed Lines (RILs) are used.
A spontaneous ‘Or’ mutant gene accumulates β-carotene in cauliflower and
the introgression of this ‘Or’ gene into Indian cauliflower resulted in the
development of β-carotene rich cauliflower) Zhang et al. (2008) discovered
SCAR markers linked to “Or” gene inducing betacarotene accumulation in
Chinese cabbage. A spontaneous mutation of a single dominant gene
designated as ‘Or’ for orange gene resulted in orange cauliflower (Dickson et
al. 1988). This Or mutant was originally observed in white curded autumn
crop cv. Extra Early Snowball. This trait was not present in Indian cauliflower.
Genome sequencing and transcriptome analysis for quality traits in
vegetables
It is a simple, low-cost, and reliable method for discovering and mapping
single-nucleotide polymorphisms (SNPs). This method is appropriate for
population studies, germplasm characterization, breeding, and trait mapping
in a variety of organisms. This procedure is based on high-throughput, next-
generation sequencing of genomic subsets targeted by restriction enzymes and
can be generalised to any species at a low per-sample cost. Next Generation
Sequencing (NSG) has reduced the cost and time for sequencing as compared
to Sanger technology and made possible to perform millions of sequencing
reactions in parallel. By combining Sanger with next generation sequencing
technologies, it is possible to sequence most crop genomes (excluding those
having very large and complex genome) with a relatively low budget.
However, transcriptome sequencing was found to be a cheaper alternative as
compared to whole genome sequencing. RNA-seq is not dependent on prior
knowledge of the sequence under investigation, allowing it to be used to
analyse poorly characterised species. It aids in the study of gene expression
and the identification of novel RNA species, as well as directly revealing
sequence identity, which is critical for the analysis of unknown genes and
novel transcript isoforms. Transcriptomic analysis has been used in
various vegetable crops for better understand of quality-related traits. Liu et
al. (2015) conducted a comparative transcroptomic analysis of genes involved
in sugar and starch biosynthesis in vegetables and discovered differential
expression of genes involved in soluble sugar and starch biosynthesis at late
development stages. Real-time quantitative (RT-PCR) analysis was used to
validate RNA-Seq data for 30 randomly selected genes that appear to be useful

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in garden pea breeding. Zhu et al. (2017) identified 797 novel genes in
watermelon to extend the available reference gene set and a large number of
differentially expressed genes (DEGs) related to plant hormone pathways,
carotenoid biosynthesis and sugar and cell wall metabolism during fruit
ripening. Transcriptomic analysis also helped in understanding of
phylogenetic relationship. Xu et al. (2015) identified ~320,000 high-quality
SNPs using transcriptome sequencing data in spinach and stated that Spinacia
turkestanica was more closely related to the cultivated S. oleracea than S.
tetrandra. Such information will be helpful in exploring and identifying wild
resources for quality improvement of vegetables. These NGS sequences are
deposited in the NCBI Sequence Read Archive, and genomes of around 40
vegetables or related species have been sequenced to date, indicating rapid
progress in the use of genomic tools in breeding vegetable crops.
Transgenic approach for quality improvement
Transgenic crops, also known as genetically modified (GM) crops, allow
plant breeders to introduce desirable genes, which were previously
unavailable, into elite cultivars, ultimately increasing their value. With the
advancement of genetics and molecular biology, a large number of quality-
related genes have been identified in various vegetable crops, including those
involved in pigmentation, biosynthesis of vitamins, minerals, and flavour
compounds, soluble carbohydrate metabolism, fruit colour, shape, size, and
position, cell wall metabolism, and shelf life. In potato, three genes, namely,
encoding phytoene synthase (CrtB), phytoene desaturase (CrtI) and lycopene
betacyclase (CrtY) from Erwinia have been introduced to produce beta
carotene. Genetically engineered potato for keto-carotenoids (including
astaxanthin) was developed by Gerjets and Sandmann (2006). To improve
flavour, nutritional content and to increase anthocyanin concentration in
tomatoes, snapdragon (Antirrhinum) genes were used (Tohge et al. 2015).
ransgenic approach also helped in extending the shelflife of vegetable crops.
For example, cytokinins are known to delay floral yellowing of plants and
Chan et al. (2009) used a transgene construct pSG766A to increase expression
of isopentenyl transferase (key enzyme for cytokinin synthesis) in broccoli.
RNA interference in vegetables for quality traits
RNA silencing is a gene regulatory mechanism that either suppresses
transcription (TGS) or activates a sequence specific RNA degradation process
[PTGS/RNA interference (RNAi)]. RNAi technology is precise, stable,
efficient and better than antisense technology and even the chances of
acceptance of products is more in public. The RNA silencing is a novel gene

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regulatory mechanism that limits the transcript level by either suppressing
transcription (TGS) or by activating a sequence- specific RNA degradation
process (PTGS/RNA interference) (Agrawal et al. 2003). In tomatoes, this
technology has been employed successfully to alter the gene expressions for
improved quality traits by increasing antioxidants (Niggeweg et al. 2004) or
suppressing over expression of negative traits such as sinapate esters in canola
(Husken et al. 2005) and alphalinolenic acid in soybean (Flores et al. 2008).
In India, there have been few attempts to use this novel technology to improve
vegetable crops in general and is used in carrot, tomato, watermelon and sweet
pepper. However, RNAi technology holds great promise for improving
vegetable crops for specific traits such as beta-carotene in tropical carrots and
late bolting in palak, radish and cauliflower, among others, as well as disease
and insect pest resistance and male sterility for hybrid seed production.
Targeted genome editing
Targeted genome editing (TGE) is a new and emerging plant-breeding
tool with a higher mutagenic efficiency than traditional breeding. Subburaj et
al. (2016) stated that targeted genome editing (TGE) technology as an
alternative tool for trait improvement in horticultural crops. The TGE
facilitates targeted and stable editing of DNA using engineered nucleases
including zinc finger nucleases (ZFNs, meganucleases, transcription
activator-like effector nucleases (TALENs) and clustered regularly
interspaced short palindromic repeat (CRISPR)/ Cas9 nucleases. ZFNs,
Meganucleases and TALENs depend on the binding and recognition of the
nucleases to specific sequences of DNA, as a result, complex engineering
processes are required to create custom nucleases that target the desired
sequence (Martin-Pizarro and Pose 2018). Li et al. (2018) reported about 5.1-
fold increase in lycopene content in tomato fruit through genome editing.
Conclusion
The nutritional qualities and associated health benefits of vegetable crops
are becoming important criteria for their inclusion in the diets of consumers.
Although vegetables are high in health-promoting compounds, but still they
lack in public health-relevant nutrients (carotene, ascorbic acid, iron, calcium,
and iodine) which needs to be improved in order to ensure their availability to
the consumers. In remote areas, where there is a problem of food shortage
during the off-season, vegetable varieties with improved nutritional values and
better storage-cum-transport life can serve much better. Biofortification could
help with this because it is a sustainable and scalable approach to combating
micronutrient malnutrition. For this purpose, breeding programs for
improving the shelf life and the nutrient quality in vegetable varieties should

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be focused. We need to find suitable donors in crop germplasm to develop
genetic stocks for specific nutrients and nutraceuticals, determine the genetics
of the target compounds in donors and create an appropriate breeding strategy,
Identify the genetics of the target compounds in donors and develop an
appropriate breeding strategy, as well as identify robust and tightly linked
molecular markers for target traits and background selection to track the level
and efficiency of introgression. But still while planning for any breeding
program for biofortified crops, some issues has to be addressed yet such as (i)
feasibility of breeding nutrient/ nutraceutical/pharmaceutical/bioactive
compounds dense varieties, (ii) target nutrient content for breeding, (iii)
chances of adoption of new genotypes by farmers (iv) impact on nutritional
status, (v) costeffectiveness of new genotypes, and (vi) consumers acceptance
of biofortified foods.
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BnSGT1 gene expression. Mol. Breed., 16: 127-138.
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19. Minoia S., Petrozza A., D’Onofrio O., Piron F., Mosca G., Sozio G.,
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 Chapter - 17
Scheduling of Irrigation for Various Major
Crops with Importance and Limitation in
Agriculture

Authors
Kalyani MSR
Ph.D. Scholar, Kerala Agricultural University, Trivandrum,
Kerala, India
Archana B
Ph.D. Scholar, Kerala Agricultural University, Trivandrum,
Kerala, India
Srinivas Y
[Link]. (Agricultural), University of Agricultural Sciences,
Dharwad, Karnataka, India
Vijaykumar P
Ph.D. Scholar, Kerala Agricultural University, Trivandrum,
Kerala, India

Page | 255
Page | 256
 Chapter - 17
Scheduling of Irrigation for Various Major Crops with
Importance and Limitation in Agriculture
Kalyani MSR, Archana B, Srinivas Y and Vijaykumar P

Abstract
Water availability is crucial for crop output sustainability which can be
achieved by improved irrigation management. Efficient management helps
farmers to schedule irrigation and reduce the amount of water without
compromising the crop yield. The main aim of irrigation scheduling is to
provide just enough water to completely saturate the plant's root zone while
avoiding overwatering, and then to let the soil dry out between waterings to
allow air to enter the soil and promote root development without overstressing
the plant. The use of additional irrigation to fill the water deficiency in the soil
has various advantages. In order to optimise irrigation, it must be possible to
supplement rainfall with adequate water at critical periods of crop growth
without overstressing the plant. Regardless of the location, the aim of water
management is to maximise productivity while satisfying crop demand. Thus,
the optimal irrigation schedule can enhance the water use efficiency by
specifying when to apply irrigation water and how much water needs to be
applied.
Keywords: Irrigation scheduling, soil moisture tension, IW/CPE ratio,
DASM and critical growth stages
1. Introduction
Agriculture sector is facing challenge to produce more food with less
water due to rising population, urbanisation and resource limitations.
Sometimes the problem grows worse due to rainfall's fluctuation. Water
management is essential for crop productivity. Two strategies are available to
address the issue of water scarcity: an engineering-based strategy (building
water reservoirs) and an agronomy-based strategy (on farm water
management). Focusing on an agronomy-based solution, we must either grow
crops that use little water or use less water on the crops we do grow. Water
resources can be managed to meet crop needs by irrigating the crop only

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during drought-sensitive growth stages and withholding water at other stages.
Therefore, developing water-saving techniques and efficient irrigation
scheduling can enhance the water use efficiency.
Irrigation scheduling is described as "the process of deciding when to
irrigate and how much water to apply, depending upon estimates or
measurements of soil water or water needed by the plant" (ASABE, 2007).
Estimates of the soil water balance or soil monitoring data are used to
determine irrigation scheduling. Some of the techniques for monitoring or
estimating the soil water status or evapotranspiration (Table 1.) includes the
hand feel and appearance of the soil, gravimetric soil water sample,
tensiometers, water balance approaches, electrical resistance blocks and
modified atmometer (Broner, 2005).
Table 1: Irrigation scheduling methods

Measured Equipment Irrigation

Method Advantages Disadvantages
parameter needed criterion
Simple, easy Low accuracy;
Hand feel Measurement
Soil to use, and sample
and of soil
Hand probe. moisture with practise, collection
appearance moisture by
content. can increase requires
of soil. feel.
accuracy fieldwork
Field work is
Obtaining labour-
Gravimetric
soil moisture Soil intensive, and
soil Auger, caps, High
content moisture delay occurs
moisture oven accuracy
through content. between
sample
sampling sampling and
results
Excellent
Reading takes
precision;
effort, requires
Tensiometers Soil immediate
Tensio- Soil moisture upkeep, and
and Vacuum moisture reading taken
meters tension breaks when
gauges tension. for soil
pressure rises
moisture
beyond 0.7 atm
tension
Salinity of the
Rapid soil has an
reading; impact; low
Moisture operates over tensions do not
Electrical Soil
resistance of Resistance a wider range make it
resistance moisture
the soil blocks of pressures; sensitive;
blocks tension.
electrically. suitable for maintenance
remote and field
reading analysis are
required

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 Depending No field work
on the model is necessary;
As it is merely
used to flexibility;
an estimate,
forecast ET, ability to
calibration and
climatic Weather predict future
Measuring regular
Water variables station or irrigation
the adjustments are
budget include readily demands;
moisture required;
approach temperature, accessible ability to
content computations
radiation, weather data schedule
are difficult
wind, many fields
without a
humidity, using the
computer
and predicted same
rainfall. equipment
Measuring It is simply an
ETo Simple to use
Modified Atmometer the estimate and
(Reference and reads
atmometer gauge moisture needs
ET). reference ET
content. calibration

1.1 Importance of irrigation scheduling

• The farmer can plan water rotation among the various fields to reduce
crop water stress and increase yields.
• By requiring fewer irrigations, it lowers the farmer's labour and water
costs while maximising the amount of moisture that can be stored in
the soil.
• By keeping deep percolation (leaching) and surface runoff to a
minimum, it minimises fertiliser expenditures.
• It boosts crop yields and crop quality, which raises net returns.
• It reduces the need for drainage, which lessens difficulties with water
logging.
• It helps manage root zone salt issues through controlled leaching.
• It generates extra revenue by irrigating non-cash crops that wouldn't
normally be watered during water-scarce periods with the "saved"
water.
1.2 Limitations of irrigation scheduling
• Small variations in cultivars, rootstocks, crop load, soil type and root
depth when combined can have a significant impact on a crop's water
needs.
• Aspects like runoff and drainage cannot be predicted or completely
understood unless plant-specific information is available.

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 • Tensiometers, gypsum blocks and capacitance probes for measuring
soil moisture are often led to malfunction, batteries run out or they
are not calibrated or placed properly.
2. Scheduling of irrigation water in major cereals
2.1 Rice (Oryza sativa)
The world's greatest population relies primarily on rice as a food source.
For one kilogramme of rice to be produced, 5,000 L of freshwater are needed.
This quantity of water has always been taken for granted, but the current global
"water crisis" puts the sustainability of the production of irrigated rice in
jeopardy.
2.1.1 Irrigation scheduling in rice
Soil matric potential may be the optimal irrigation criterion, since soil
texture, cultural practises, atmospheric vapour pressure deficit and water
management all affect the amount of water needed for rice irrigation. The basis
for timing irrigation for rice and other crops can be found in the measurement
of soil matric potential using inexpensive, locally made tensiometers (Hira,
1993; Shock et al., 2002). Tensiometer helped with irrigation rice with better
irrigation planning can use less irrigation water without losing yield. The
tensiometer helps conserve energy (70-101 kWh/acre) and water of about 14-
15 per cent (Vatta et al., 2013).
[Link] Phasic stress irrigation
Irrigation under phasic stress can be used on summer rice in areas with
limited water supplies to save a significant amount of irrigation water without
significantly lowering output. The similar water resources can irrigate about
20-30 per cent more land. The amount of water saved varies between 24 and
36 per cent of the required for 5 cm of continuous submersion throughout the
crop growth, depending on the following schedule (Table 2.). It only reduces
grain yield by 0.1 to 1.6 percent.
Table 2: Categories of irrigation schedule for rice
Stages
ScheduleRooting to maximum Maximum tillering to Heading to
tillering heading maturity
Category I Continuous submergence Saturation point* Saturation point*
Continuous
Category II Saturation point* Continuous submergence
submergence
Hair cracking of
Category III Continuous submergence Continuous submergence
surface*
Hair cracking of Hair cracking of
Category IV Continuous submergence
surface* surface*
*Irrigation at 5 cm to be given at the stages marked.

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2.2 Wheat (Triticum aestivum)
Wheat is the second most important food crop in India. Well drained
loams and clayey loams are best suited with cool and sunny winters are very
conductive for growth of wheat. Wheat grows well in areas receiving an
annual rainfall about 25-175 cm and regions. Crop requires 4-8 irrigations
depending on soil type and amount of rainfall. Irrigation at proper time and at
appropriate physiological stages is must for optimum wheat yield.
2.2.1 Irrigation schedule
• First irrigation: 20-25 DAS, at the crown root stage.
• Second irrigation: 40-45 DAS, at the tillering stage.
• Third irrigation: 60-65 DAS, or at the node formation stage.
• Fourth irrigation: 80-85 DAS, or at the blossoming stage.
• Fifth irrigation: 100-105 DAS, at the milk formation stage.
• Sixth irrigation: 115-120 DAS, or at the grain filling stage.
Four irrigations at the following stages can produce a good yield in the
case of loam and heavy loam soils:
• 20-25 days after planting, begin irrigation.
• After 30 days, apply the second irrigation.
• 30 days following the second irrigation is when the third irrigation is
applied.
• 20 to 25 days following the third irrigation, the fourth irrigation.
In case of limited irrigation resources:
• The crown root stage, spike emergence stage, and milk formation
stage should all receive irrigation if there are only three irrigations
available.
• If there are only two irrigations available, use them at the crown root
and flowering stages.
• Irrigate the crop during the crown root stage if there is only room for
one irrigation.
• Irrigation depth should be 6 cm for light soils and 8 cm for heavy
soils, respectively.
2.3 Maize (Zea mays)
Rabi corn needs 450-600 mm of water while Kharif maize only needs
400- 550 mm. Despite the fact that maize roots can extend up to 120 cm, their

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density is highest at 60 cm. The first irrigation should be applied up to a depth
of 120 cm. The timing of the second irrigation depends on the soil, the weather
and the wind speed. In dry weather, space out irrigations more frequently; in
humid weather, space them out less frequently. Planning irrigations in maize
to deplete the field's capacity of 50% of the soil moisture is advantageous.
[Link] Irrigation schedule for Kharif maize
• During the kharif season, when rainfall is evenly distributed and
sufficient to provide adequate soil moisture throughout the crop's life
cycle, the crop is cultivated through rain-fed farming.
• In order to build high and stable yields, the available irrigation
sources should be used to supply one or two irrigations at the crucial
stages of any dry spell period.
• Soil moisture stress and water logging are known to be problems for
maize.
• By providing surface drainage, sowings made on ridges can get
around the problem of uplands becoming flooded.
• Throughout its life cycle, maize is vulnerable to moisture stress. If
rain does fall, irrigation should be applied to the crop during the most
crucial flowering and grain-filling periods.
[Link] Irrigation schedule for Rabi maize
• Insufficient rabi rainfall is the main drawback for the effective
development of high-yielding maize hybrids.
• One of the key elements influencing the success of a crop is the
timely availability of reliable irrigation.
• In heavy soils, a moisture level of 30% during the vegetative stage
and 70% during the reproductive and grain-filling period is desirable
for obtaining the maximum yield.
• For the best yield in heavy dense soils, 30% moisture level during the
vegetative stage and 70% moisture level during the reproductive and
grain-filling stage is preferred.
• During the rabi crop season, four to six irrigations are necessary. The
following crop growth phases should be utilised if six irrigations are
given:
a) Two irrigations (20-25 days apart) up until flowering
b) One irrigation (important) at flowering

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 c) Two irrigations after flowering
d) One irrigation at the beginning of grain filling.
• One irrigation during the vegetative stage may be skipped if only five
irrigations are given, and irrigation may be supplied after the dough
stage if only four irrigations are given. However, if sufficient rainfall
is received, the irrigation schedule may be adjusted appropriately.
3. Scheduling of irrigation water in major millets
1.1 Sorghum (Sorghum Bicolor)
It is a sturdy crop and can withstand varied climatic hazards more than
any other crop. It is primarily a tropical warm weather crop. It can withstand
wide range of temperatures varying from15.5°C to 40.5°C with rainfall
variations of 35-150 cm per annum. Sorghum can be grown in heavy black
soils to red soils, but comes up well in heavy black soils. The best crops are
grown the clay loams. Jowar does not grow well under gravely and marshy
soils.
1.1.1 Irrigation scheduling: Sorghum can be planted either through
transplantation or through the direct seeding method. According to the
method opted the irrigation strategy changes.
a) Transplanted sorghum
1) Nursery: In case of the transplanted sorghum irrigation at the
nursery stage is crucial for the establishment of the crop. Usually,
four irrigations are suggested first irrigation at immediately after
sowing; second irrigation at 3rd day after sowing; third sowing at 7th
day after sowing and fourth irrigation at 12th day after sowing in case
of the red soils whereas, in case of heavy soils irrigation at
immediately after sowing 4th, 9th, 16th day after sowing is opted for
better growth.
2) Main field: Immediately after transplanting irrigation has to be given
on 1st and 4th day for better establishment; irrigating at 15th and 28th
day after transplanting will help for the growth of the plants at
vegetative stage; at flowering which coincides with the 40th and 52nd
day after transplanting irrigation is mandatory in order to boost the
growth of economical parts; one irrigation at maturity phase at 65th
day after transplanting will help translocation of metabolites to the
sink.

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b) Direct sown sorghum: In direct sown sorghum irrigation immediately
after sowing and after 4th day after sowing will promote the better
germination of the seed; irrigation at 15th and 28th day after sowing
coinciding the vegetative stages of the crop; at flowering stage irrigation
on 40th, 52nd, 64th day after sowing is crucial followed by the irrigation at
76th and 86th days after sowing at maturity is important for maximizing
the yield. The peak water need of the crop is at booting to seed setting
stage. Irrespective of the method of planting irrigation can be given
through critical stage approach at germination (at the time of sowing),
Kknee high stage (30-35 DAS), flag leaf stage (50-55 DAS), flowering
(70-75 DAS) and grain formation stage (100-105 DAS).
1.2 Bajra (Pennisetum americanum)
Pearl millet is endowed with greater ability to withstand harsh climatic
factors and still yield substantially. The crop requires low annual rainfall
ranging between 40 to 50 cm and dry weather. The crop needs wet weather,
light showers and bright sunshine, at its grand growth stage. There should be
no rain at flowering as the pollen will be washed off and fertilization would
be poor, while at maturity it needs fairly dry weather associated with bright
sunny days. The crop may tolerate drought but cannot withstand high rainfall
of 900mm or above.
1.2.1 Irrigation scheduling
• Total water requirement is 450-550 mm. Generally, two irrigations
during the growing period of the crop are enough.
• If moisture is limiting, irrigation must be given at the time of ear head
emergence because it is the most critical stage for moisture stress.
• Bajra does not tolerate water logging. Proper arrangement for
draining the excess water must be made.
1.3 Finger millet (Eleusine coracana)
Finger millet is an important minor millet grown in India. It is a staple
food crop in many hilly regions of the country. In fact, it is the main cereal
crop for monsoon season in some hilly areas. It is a hardy crop. In regions of
higher rainfall, it can be raised on well drained soils as a transplanted crop. It
can be grown on a wide variety of soils ranging from very poor to very fertile
soils. It thrives best on well- drained loam or clay loam soils. It can tolerate
salinity better than other cereals. Finger millet is not a season bound crop. So,
it can be grown throughout the year, if water is available.

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1.3.1 Irrigation scheduling
• Transplanting is done under assured irrigation. 1) Ridges and furrows
2) Flatbed method. Seeds are raised in nurseries and after 20 days,
seedlings are ready for transplanting in main field. Usually, 5
irrigations are preferred during the nursery stage in light soils with
three to four days interval and 4 irrigations in heavy soils with five
days interval.
• After establishment of seedlings for a week or 10 days, irrigation
should be cut off. This helps in hardening of seedlings and for
vigorous and healthy growth.
• Ragi sown during Kharif, generally does not need any irrigation.
Adequate moisture should be provided at the time of flowering and
grain setting stages.
• It cannot tolerate waterlogged conditions. Hence, drainage facilities
should be provided.
• Critical stage irrigation approach under water scarcity by irrigating
at establishment stage (1-7 days); vegetative phase (8-20 days);
flowering phase (21-55 days); maturity phase (56-120 days) should
be followed.
1.4 Proso millet (Panicum miliaceum)
The crop is able to evade drought by its quick maturity. Being a short
duration crop (80-90 days) with relatively low water requirement, it escapes
drought and, therefore, offers better prospects for intensive cultivation in dry
land areas. It is a crop of warm climate. It is highly drought resistant and can
be grown in areas where there is scanty rainfall. It can withstand water
stagnation also to some extent. It is highly drought resistant and can be grown
in areas where there is scanty rainfall. It can withstand water stagnation also
to some extent.
1.4.1 Irrigation scheduling
• Generally, does not require any irrigation. However, at tillering stage,
if dry spell prevails for longer period, then one irrigation must be
given to boost yield.
• Summer crop however would require 2-4 irrigations depending upon
soil type and climatic conditions. Give first irrigation 25-30 days
after sowing and second irrigation about 40-45DAS.

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4. Scheduling of irrigation water in major pulses
4.1 Pigeonpea (Cajanus cajan)
The prolonged duration of the kharif pigeonpea crop, which lasts until
January or February, may make it the only pulse crop to receive irrigation
(postrainy season). The kharif crop is irrigated for another reason: it produces
two or three flushes of harvests over the course of six to seven months. One
irrigation at flower commencement and another during pod development can
increase the yield of a crop grown throughout the rainy season in the
nonappearance of timely rainfall. Post-rainy season crops respond to watering
more consistently because to the moisture in the soil that has been retained.
On light soils, three irrigations spaced out monthly can treble the yield; but,
on medium as well as deep soils, more than two irrigations may not be required
to maintain the crop during the post-rainy season.
4.1.1 Irrigation scheduling in pigeonpea during postrainy season
• Two irrigations, one at pod development and the other at flower
initiation, can quadruple the output of a kharif crop in the absence of
sufficient rainfall after October.
• Post-rainy season crops respond to watering more consistently
because to the moisture in the soil that has been retained. On light
soils, three monthly irrigations can treble the yield, while on medium
as well as deep soils, more than two irrigations may not be required
during the post-rainy season.
• In order to achieve the best yield, irrigation should be planned with
0.75 (IW/CPE ratio) during the post-rainy season.
• If irrigation is planned according to DASM, three irrigations at a level
of 75% depletion can be sufficient for the best yield.
4.2 Chickpea (Cicer arietinum)
Approximately 60% of the country's chickpea acreage is rainfed,
particularly when grown on black soils after the rainy season when soil
moisture is receding. The crop reacts to irrigation in north India's light soils,
especially if the winter rains are insufficient. Not more than two irrigations are
required, not even during drought circumstances. Irrigation can quadruple the
yield on rather light textured soils in areas with high evaporative demand, such
as Peninsular India. However, irrigation promotes excessive growing and
lodging when evaporative demand is low, as it is in north India. The essential
stages for stress caused by soil moisture are branching at 45 DAS and pod
development at 75 DAS.

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4.2.1 Irrigation scheduling for growing chickpea
• On light soils, it appears that four irrigations- for sowing, branching,
flowering and pod filling are sufficient if irrigation water is available.
Irrigation is typically not advised in case of deep black cotton soils.
• Two irrigations at branching and pod development can produce an
economically viable yield on light soils in the case of restricted
availability.
• Planning irrigation at a ratio of 0.5 IW/CPE or at 75% DASM,
needing 2 to 3 irrigations, is just as beneficial as irrigation at key
times.
• The sensitivity of chickpea to soil salt. The yield can be reduced by
60% when using irrigation with saline water having 10 mmhos cm-1
concentration.
4.3 Greengram (Vigna radiata) and Blackgram (Vigna mungo)
During the kharif season, greengram and blackgram are grown as rainfed
crops. As a result, unless there is a lengthy dry period at a critical point for soil
moisture stress, they won't respond to watering. Additionally, no irrigation
will be used to post-rainy season crops produced on black soils with stored
moisture. In the north of the country during the summer, and in the south
during the winter, these crops are farmed as irrigated crops. These crops are
produced as relay crops alongside kharif rice in delta regions of south India
without any irrigation. Flowering in addition to pod development periods are
vulnerable to a dry soil condition. For a normal yield in various sites, summer
crops needed three to four irrigations at an IW/CPE ratio of 0.60.
4.3.1 Irrigation scheduling for greengram and blackgram
• For the best production during the winter and summer seasons, four
irrigations- one each during sowing, branching, flowering, and pod
development- are sufficient when there are appropriate irrigation
facilities.
• If available irrigation water is limited, pre-sowing irrigation and a
second watering during flowering can produce crops with yields that
are nearly as high as they can be.
• When water resources are inadequate, scheduling irrigation at 50%
DASM (blooming and pod formation) and at 75% DASM (other
stages) may be just as successful as the method described above.

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 • Unless there are extended dry spells that occur during the crucial
stages of soil moisture stress, it may not be necessary to irrigate these
crops throughout the rainy season.
Other irrigated pulse crops, such as cowpea, kidney bean and lentil, are
not very significant. The irrigation technique for these crops is the same as for
blackgram and greengram.
5. Scheduling of irrigation water in major oilseeds
5.1 Groundnut (Arachis hypogaea)
Groundnut is an important source of oil and protein to a large portion of
the population in Asia, Africa and the America. It is a self -pollinated, annual,
herbaceous legume originated from South America. Usually grows in areas
where the rainfall exceeds 500mm in a growing season. It can be grown in
wide range of soils yet best suited for the sandy soils. It is considered as the
most acid tolerant crop as it can withstand the pH range of 5-5.5. It can be
grown in all the three seasons but the yields will be high if grown in summer
with adequate water.
5.1.1 Irrigation scheduling
• Usually, the crop is cultivated in the rainfed kharif season.
• Early growth phase till 20 days after sowing is the least sensitive
stage to the moisture.
• Flowering, peg penetration and pod development are the three critical
stages for the irrigation.
• Scheduling irrigations at 25% DASM throughout the growth period
results in high pod yield in sandy soils.
• Irrigating the crop at
o 25% DASM- from pegging to early pod development
o 50% DASM- at other stages appear to be ideal for high WUE
without significant reduction in pod yield.
• When first irrigation is given at 25 DAS, which creates moisture
stress in the soil that promotes good root system, reduces excessive
vegetative growth, encourage the better nodulation, induces high
flowering.
• Under water deficit situations, irrigation at 25 DAS; followed by two
irrigations at 15 days interval between 15 and 75 DAS will minimize
the yield loss.

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 • When water is not limiting pre-sowing, irrigation followed by an
irrigation at 25 DAS, four irrigations at 10 days interval and final two
irrigations at 15 days interval will improve the yield.
5.2 Sesame (Sesamum indicum)
Sesame is a short-day plant and is temperature sensitive at low
temperature of 100 C and below growth retardation takes place and when the
temperatures are above 400 C pollination gets affected and the capsule
formation is reduced. Optimum temperature for growth of sesame is 25-270 C.
5.2.1 Irrigation scheduling
• Water requirement of sesame is 300-350 mm. For Rabi season crop
5-6 irrigations are required preferably applied at sowing followed by
3rd, 6th, 8th, 9th weeks after sowing.
• In case of no rain/scarcity in irrigation water, critical stage approach
where the irrigation at pre-sowing of the crop, at flowering and
capsule filling stages has to be given to secure the economical parts.
• Irrigation at 50% DASM and through IW/CPE ratio method at a ratio
of 0.6 is optimum.
5.3 Sunflower (Helianthus annus)
It is the most important oil seed crop in India. It is the most preferred
edible oil in the southern parts of the country. It is a photo insensitive crop
which makes it suitable to be cultivated in all the three seasons of the year.
Wide adaptability to different soils, its short duration (90-100 days) and high
productivity, tolerance to drought and saline conditions, high economic value
makes the farmers attracted to cultivate the crop.
5.3.1 Irrigation scheduling
• Sunflower is an irrigation responsive crop and is an indicator plant
for moisture stress with the water requirement of 500-600 mm.
• It has deeper root system which makes the crop to withstand short
periods of drought, this result in the stable yield of the crop even
under low rainfall. It also has physiological plasticity to adapt to the
available moisture present in th soil to complete its life cycle.
• The critical stages approach for irrigation is at Bud initiation (30
DAS), flower opening (45-50DAS) (most sensitive to moisture
stress) and seed filling (60-75 DAS), central portion of the head
remains unfilled under moisture stress conditions.

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 • During kharif the crop needs 3-6 irrigations and in rabi 4-8 irrigations
and irrigation should be curtailed 20 days before the harvesting.
• Through DASM method of irrigation 50% DASM is followed in all
the growth stages excluding the critical stages where the irrigation is
preferably given at 30% DASM.
• Through IW/CPE ratio method the irrigation is ideally given at a ratio
of 0.5- 1.0.
5.4 Rapeseed and Mustard (Brassica sp.)
Rapeseed and mustard are the predominant rabi season oilseed crops in
India. India is one of the largest producers of the rapeseed and mustard after
China. Rapeseed (Brassica compestris var. Sarson and toria) is easily
distinguished from the Mustard (Brassica juncea) with its short height, leaves
are born sessile and are glabrous and hairy, fruits are thicker and are laterally
compressed with a beak of one- third to half their length. The crop requires
relatively cool temperature and a dry harvest period. The crops are long day
plants which does not possess any tolerance to drought or water logging.
5.4.1 Irrigation scheduling
• Water requirement of the crop ranges between 450-600 mm.
• The brassicas even with less water requirement they are considered
as the water responsive crops.
• Usually, they utilize the residual moisture present in medium to
heavy soils for their growth.
• Delayed first irrigation is recommended in brassicas which helps in
profuse branching, flowering and fruiting.
• Through critical stage approach pre-sowing irrigation, irrigation at
flowering and at fruiting stage is beneficial.
6. Scheduling of irrigation in major commercial crops
6.1 Cotton (Gossypium sp.)
Due to its strong root system, cotton is a crop that can withstand drought.
The crop needs 600 to 800 mm of water. Cotton cannot tolerate excessive soil
moisture; thus, it does not require frequent irrigation. The time between two
irrigations is influenced by the soil type, the amount of rainfall, and other
meteorological factors. During the flowering and fruiting stages, the crop must
not be allowed to experience water stress; otherwise, excessive flower buds
and young bolls may fall off, reducing the crop's production. At any stage of

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growth, the crop cannot withstand conditions of water logging. Square
creation, flowering, and boll development are the three critical stages.
6.1.1 Irrigation scheduling
• In case of row cropping first irrigation is given after 30-35 DAS and
later after a gap of 10-12 days.
• In case of cotton-wheat cropping system, last irrigation is given till
10 October.
• In case of furrow beds first irrigation is at 2-4 days old seed sowing,
second after 6-9 days and subsequent irrigation should be given at 15
days interval. Irrigation scheduling can be changed according to
prevailing weather conditions, water availability and crop growth
stages.
• If the seedbed is too dry to support germination, pre-planting
irrigation is preferable to irrigating after planting.
• Some water deficits early in the season can stimulate root production
and development of a deeper root system, especially in sandy soils
• Crop water demand increases from 0.2 to 0.28 inches per day during
first flower to peak bloom period
• Once cotton plants have emerged and are growing, irrigation should
be used to supplement rainfall and prevent depleting the soil moisture
profile prior to bloom
• Growers should reduce or stop irrigation and allow plants to
experience some water stress after bolls start opening. This stress
reduces regrowth, makes defoliation easier, and encourages boll
opening.
6.2 Sugarcane (Saccharum officinarum)
Sugar industry is the second largest industry after textiles in India. In India
sugarcane is grown in both tropical and sub-tropical regions. The crop prefers
long sunshine hours and high relative humidity. It is a highly water
consumptive crop, so mostly suitable for high rainfall areas but if the rainfall
coincides with the maturity phase of the crop the quality and quantity of the
juice.
6.2.1 Irrigation scheduling
• To produce one tonne of sugarcane the crop consumes about 250
tonnes of water.

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 • The crop is highly sensitive to moisture stress under severe stress the
crop may lose yield upto 60-70 percent.
• It is considered that the irrigation requirement of the crop cannot be
reduced without compromising on the yield
• In tropical region it is preferred to irrigate once in 7 days during
germination stage which would fall between 1-35 days after planting,
at 10 days interval during tillering stage (36-100 DAP), irrigating at
7 days interval during grand growth stage (101-270 DAP), at 15 days
interval during maturity stage (271 DAP to harvest) has to be given
according to the rainfall pattern. On an average sugarcane needs 30-
40 irrigations till harvest.
• In early planted crop, moisture stress will be experienced at grand
growth stage which affects the stalk elongation.
• In late planted crop the moisture stress coincides the formative phase
which effects maturity and ripening of the cane.
• At ripening stage, care should be taken that the moisture stress should
not be present as it affects the sugar content.
• Growing sugarcane in waterlogged situation is also a disadvantage as
it reduces the sugar recovery.
• Light and frequent irrigations will give higher yield.
• Through IW/CPE ratio approach at a ratio of 0.75-0.50, irrigation can
be provided during tillering, grand growth and maturity phases.
7. Conclusion
Effective irrigation scheduling reduces runoff, leaching and percolation
losses, which maximises irrigation efficiency by utilising water and energy
more effectively. Because excessive soil moisture frequently causes plant
disease, nutrient leaching and diminished pesticide effect, over irrigation can
lower yields. Crop failure is often caused by inadequate irrigation. Proper
irrigation schedule saves water without lowering production. Therefore,
effective irrigation scheduling helps maximum profit and maximise water and
energy use.
8. References
1. ASABE, 2007. Soil and Water Terminology. American Society of
Agricultural and Biological Engineers, St. Joseph, Mich.
2. Broner, I., 2005. Irrigation Scheduling. Bulletin 4.708. Colorado State
University, Ft. Collins, Colorado.

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3. Crookston, B., Blaster, B., Darapuneni, M. and Rhoades, M. 2020. Pearl
millet Forage water use efficiency. Agronomy. 10: 1672.
4. Hira, G.S. 1993. Locally fabricated tensiometers, their problems and use.
In: Proceedings, national seminar on developments in soil science. Indian
Society of Soil Science, New Delhi, pp. 455-456.
5. ICAR- Sugar cane breeding institute.
[Link]
production?id=340&phpMyAdmin=11c501a2a5dt8788ed6#:~:text=In%
20tropical%20area%2C%20irrigations%20are,during%20maturity%20p
hase%20(271%20days
6. Irrigation schedule for sesame cultivation; Agriculture experts.
[Link] [Link]
sesame-cultivation-agriculture-
experts/#:~:text=Usually%203%2D4%20irrigation%20are,days%20afte
r%20sowing(DAS).
7. Ismail, S.M. 2012. Optimizing productivity and irrigation water use
efficiency of pearl millet as a forage crop in arid regions under different
irrigation methods and stress. African Journal of Agricultural Research.
7(16): 2509-2518.
8. Murugu, F.S., Nyamudeza, P., Mugabe, Francis., Matimati, I. and
Mapfumo, S. 2006. Effect of seedling age on transplanting shock, growth
and yield of pearl millet (Pennisetum glaucum L.) in semi-arid
Zimbabwe. Agronomy. 5(2).
9. Natarajan, R. and Wahab, K. 2001. Influence of irrigation and mulching
practices on water use, yield components and yield of finger millet
(Eleusine coracana). Indian Journal of Agronomy. 46(4): 702-706.
10. Shock, C.C., Feibert, E.B.G., Seddigh, M. and Saunders, L.D. 2002.
Water requirements and growth of irrigated hybrid poplar in semi-arid
environment in Eastern Oregon. Western Journal of Applied Forestry.17:
46-53.
11. TNAUagriportal
[Link]
~:text=Water%20Management&text=Irrigation%20should%20not%20be
%20delayed,70%20%2D%2080%20days%20after%20ratooning
12. Vatta, Kamal, M.S. Sidhu, and Kaur, A. 2013. Pattern of growth and
emerging challenges for Punjab agriculture, Report submitted to the
Unites Nations Environment Programme (UNEP), Punjab Agricultural
University, Ludhiana, India.

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Page | 274
 Chapter - 18
Precision and Regenerative Agriculture

Authors
Asha
XXX
Diksha
XXX
Shabnam
XXX
Priyanka Sanwal
XXX

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Page | 276
 Chapter - 18
Precision and Regenerative Agriculture
Asha, Diksha, Shabnam and Priyanka Sanwal

Historical background
The concept of precision agriculture has been emerged first in 1980s in
the United States. The farmers in America were trying to maximize the
production by investing the money only in areas where there is need of
fertilizers. This idea rises the concept of adding fertilizers with varying rate
according to the need as identified the GPS guided Grid or zone sampling.
Therefore, the need of fertilizers was optimized. Around the nation, the
development of precision and regenerative agriculture developed at varying
time period and the leading countries in this step were United States, Canada
and Australia. The use of precision concept in livestock farming resulted in
better monitoring of animal productivity, human and environmental health.
Introduction
Agricultural sector has embraced the use of precision agriculture to gain
production and environmental and economical benefits. As this is the new and
evolving area to study, several definitions have been provided for the precision
agriculture. It may also be called as site-specific crop management, smart agri-
culture or information-rich agriculture. Precision agriculture, in wider sense,
is the application of management decisions in space and time based on
identifying, quantifying and responding to variability. Precision
agriculture (PA) is the management strategy that is based on observing,
measuring and responding to temporal and spatial variability to sustain the
agricultural production, and that can be used for both crop and livestock
production. The precision agriculture research is aimed at optimizing returns
on inputs with preservation of resources by working as a decision support
system (DSS) for whole farm management. The precision agriculture is highly
linked with the sustainability. The term ‘sustainability’ was originally used to
refer to agricultural and industrial technologies that reduced the environmental
degradation which are associated with economic activity.

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 Doing right things at right time and right place is the basic concept of
precision agriculture (Bongiovanni and Lowenberg, 2004)/site-specific
management (SSM) fig 1.

Fig 1: Sustainability as described by the intersection of three disciplines: Ecology,

economics and sociology.
Precision agriculture opens the way for site-specific management with the
help of information technology and therefore makes it practically feasible for
commercial agriculture. The pecision agriculture involves the all production
practices of agriculture which rely upon information technology to obtain the
excepted outcomes and also to monitor those outcomes such as yield, variable
rate application (VRA). The site-specific management (SSM) is defined as the
electronic monitoring and control that are applicable for data gathering,
information processing and decision support for the temporal and spatial
allocation of inputs for crop production (Lowenberg-Deboer and Swinton,
1997). Their main focus was on agronomic crops, but some contradicts were
also there regarding production of horticultural crops and livestock
management. Furthermore, reliable detection, accurate identification and
proper quantification of pathogens and other factors affecting both plant and
animal health, are crucial to be kept under provision in order to minimize
economic expenditures, trade disruptions and even human health risks.
Precision farming and measurements have already fixed blueprint in order
to increase farm productivity and quality, as well as improving working
conditions through reduction of manual labour. All these factors are essential
for sustainable farm production. Also, several modern farmers already use
scientific solutions, e.g., digitally-controlled farm plants and also unmanned
aerial vehicles (UAVs) for monitoring and forecasting. Daponte et al. (2019)
studied the use of drones in precision agriculture as drones are available at

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reasonable prices and are capable of imaging ground data with corresponding
geographic locations to have a complete and clearer picture of the ground
information. Drone images and ground sensor data will provide wide room for
scientific research and development. Furthermore, several meteorological
aspects have to be considered for developing such platforms, from the sensors
embedded on them up to the instrumentation and the calibration procedures
for their testing. Regardless of their effectiveness and usefulness, the main
flaw lies on the fact that these systems are calibrated only for a specific task
(e.g., classifying different kinds of vegetation, water bodies, urban, bare soil,
etc.), without the ability of creating a holistic view of agricultural processes.
This lack of interoperability causes additional work for the human operators,
since they have to manually feed the output data from one system to another.
For all such reasons, software modules, drones and other equipment are object
of research in order to develop a common information middleware and
application interface. The aim is to reduce monotonous and time-consuming
work.

Fig 2: The monitoring operations of a crop by means of multi-spectral camera

embedded on drone (Source: Daponter et al., 2019).
Major components of precision agriculture
The major tools of precision agriculture are Global Positioning System
(GPS), geographic information system (GIS), miniaturized computer

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components, mobile computing, in-field and remote sensing, automatic
control, advanced information processing, and telecommunications etc. But
including GPS, GIS, Seelan et al. (2003) has also involved yield monitoring
devices, soil, plant and pest sensors, remote sensing, and variable-rate
technologies (VRA) for applicators of inputs in PA tools.

System elements of precision farming

A) Global Positioning System (GPS)
Global positioning system has revolutionized positioning concept, though
it started primarily as a navigation system. Today, the Global Positioning
System (GPS) possess wider utility on national as well as international level.
Despite of its ease of use and worldwide all-weather operation, GPS owes its
popularity to the dependable high accuracy with which position, time and
direction can be determined. (Ajai, 2002 from shanwad et al). As a component
of precision Agriculture, Global Positioning System satellites broadcast
signals that allow GPS receivers to calculate their position. This information
is provided in real time, meaning that continuous position information is
provided while in motion. Having precise location information at any time
allows crop, soil and water measurements to be mapped. GPS receivers, either
carry to the field or mounted on implements allow users to return to specific
locations to sample or treat those areas.

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 Global Positioning System is a satellite-based radio navigation system
(with a complete set of at least 24 satellites orbiting the around earth in a
defined pattern) that provides accurate 3- dimensional location data (latitude,
longitude, and elevation) data worldwide at any time, in any weather. The GPS
works with two modes, one is, single receiver mode which collects the timing
information, timing is processed into position; and another one is differential
mode (DGPS) that utilize two receivers out of which one receiver is elevated
in a stationary position and the other is on the machine/implement. The
farmers are able to collect soil samples from a specific location in the field
accurately, every year, to monitor crop conditions so, the macro- and micro-
scales spatial variability of soils could be observed using GPS navigators.
Inspite of this, the complete planning (field boundaries, acreage for field areas,
roads, irrigation systems, distances between points and weed and disease
affected areas) could be generated by scrutinize the agricultural land.

Fig 3: GPS segments

B) Geographical Information System (GIS)
A geographic information system (GIS) is a computer system for
capturing, storing, querying, analyzing, and displaying Geo-spatial data
(Chang, 2014). Geospatial data explains both the location and the attributes of
spatial features. For example, to describe a road, its location (i.e., where it is)
and its attributes (e.g., length, name, speed limit, and direction) as GIS has the
capacity to store and analyze large amount of data at high speeds. A GIS
permit the user to manage road data and many other geospatial data, thus
differentiating it from business management systems that deal with non-spatial

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data. Geographic information system is considered as the brain of Precision
Agriculture. A GIS comprises hardware, software, people, and organization,
in addition to geospatial data. GIS hardware includes computers for data
processing, data storage, and input/output; printers and plotters for hard-copy
maps; digitizers and scanners for digitization of spatial data; and GPS and
mobile devices for fieldwork. The technical method for studying variability
aspects of a piece of land is through interpolation technique. GIS operations
usually involve data acquisition, data management, data query, vector data
analysis, raster data analysis, and data display. GIS has been important in
resource management, land-use planning, natural hazards assessment, wildlife
habitat analysis, riparian zone monitoring, and timber management. GIS has
also been used for emergency planning, crime analysis, public health, land
records management, transportation applications, precision farming, and
military operations. GIS maps are helpful to evaluate environmental health
also e.g. ambient air pollution study.
C) Remote sensing
Remote Sensing (RS) is the science of obtaining and interpreting
information from a distance, using sensors that are not in physical contact with
the object being observed (Jensen, 1996). The science of remote sensing
includes aerial, satellite and spacecraft observations of the surfaces and
atmospheres of the planets in our solar system, while the Earth is noticeably
the most frequent target of study. Remote sensing is often restricted to
methods that detect and measure electromagnetic energy including visible and
non-visible radiation that interact with surface materials and the atmosphere.
Yie et al. (2019) have divided RS system components in to three broad
categories namely spatial foundation system, ground base system and remote
sensing data storage system.
• This technology is beneficial in getting information about earth’s
surface without any destruction.
• It systematically obtains information about large geographical data.
• It is beneficial in getting data about locations which are not accessible
to human interventions.
• The data obtained through remote sensing is independent of other
technology such as GIS.

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 Remote sensing process
Remote sensing technology has been used as a key component of site-
specific management/precision agriculture and is being used by an increasing
number of scientists, engineers and large-scale crop growers. During the last
two century, evolution in RS data collection capabilities, data processing and
interpretation of ground based, airborne and satellite observations have made
it possible to combine RS technologies and precision crop management
systems (Waheed et al., 2006). Currently, there is a wide range of satellite data
that differs in (i) technique (active/passive, radiomete), (ii) spatial resolution
from sub meter to kilometers (iii) spectral range, and (iv) viewing geometry
(Oza et al., 2008). Different types of sensors such as multispectral and
hyperspectral sensors are able to generate large amount of data in effective
manner by using remote sensing. Kurtza et al. (2009) used multi-temporal
Landsat imagery in order to classify land cover types and grazing intensity.
The use of remote sensing in agriculture has led to making a number of policy
level decisions for food security, poverty alleviation and sustainable
development using RS imagery. Worldwide mapping of land use, land cover,
soil and wasteland have helped in expansion and intensification of agricultural
activities and also in identification of land capability classes and crop
suitability indices (Venkataratnam, 2002). Although the remote sensing
cannot capture all types of agricultural information, it can possibly give
accurate and timely information to guide agronomic and economic decision-
making.
D) Variable Rate Technology (VRT)
Addition of inputs precisely in variable type and quantity at different sites
of field/area according to the present status and requirement is the practice of

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variable rate technology (VRT) in precision agriculture. This is highly
beneficial for successful crop production with low-inputs. In VRT (variable
rate technology) and VRA (variable rate applications), in conjugation with
sprayers are available to deliver the correct amount of chemicals depending
on farm conditions like crop growth stage, crop condition and previously
collected crop growth data. It involves two types of methods basically which
are map based and sensor-based.
In case of map-based VRT, the application rates are adjusted on basis of
electronic/ prescription maps. By knowing the field position from a GPS
receiver and a prescription map of desired rate, the concentration of input is
adjusted as the applicator moves through the field.
Whereas the sensor-based VRA don not need map or positioning system.
Sensors that are present on the applicator will evaluate the soil properties or
crop characteristics. Based on this collected information, a control system then
measures the input needs of the soil or plants and transfers the information to
a controller, which delivers the input to the location measured by the sensor.

Fig 4: VRA spraying system that is a flow-based control system of application rate.
The VRA is able to improve economic return by optimizing inputs in each
management zone. VRA allows us to focus inputs on management zones that
provide the highest return, while reducing inputs in lower productivity zones
or where previous management has resulted in a situation for reduced input
need.

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Soil monitoring and yield maps
A yield map is a useful starting point for visualizing and locating areas of
variation. Modern grain harvesters are generally equipped with a yield monitor
capable of logging yield information every 1-2 second. The yield map can
provide the basis for locating areas in which to take soil samples and can also
be used to determine replacement nutrient maps for the following crop. While
a yield map provides a useful synopsis of variation, it gives no indication of
the cause of the variation.
Optical satellite imagery is widely available and can also provide a
valuable starting point for locating changes in soil properties, especially color,
which can relate to soil texture and class. Proximal tools for on-the-go soil
sampling continue to evolve from those measuring a single parameter, for
example, pH or organic matter, to multiple physical and chemical
characteristics being recorded in a single pass. A widely used on-the-go soil
sampling method has been to measure electromagnetic induction. This can be
done aerially but for agriculture is generally done proximally using a
transmitting and receiving coil in contact with the soil surface.
Regenerative agriculture
The continuously increasing population is putting a pressure on global
food system. The over-dependence on fertilizers, soil degradation, loss of
biodiversity, ecosystem imbalance etc. has led to various ill effects. All these
aspects have given rise to regenerative agriculture, or in other words,
regenerative farming. The word ‘regenerative’(adjective) is also called for
renewal, transformation and revitalization of the global food system by civil
society groups and corporations (Duncan et al., 2021). From this context, the
use of soil, plant and ecosystem should be in sustainable manner. Now the
question arises that what are the evidences that motivates to proceed towards
the regenerative agriculture and what are the agronomic solutions to approach
these evidences. In view of this concept, some perceptive that mainly focuses
on regenerative agriculture are emerging food crisis. The terms Regenerative
Agriculture and Regenerative Farming came into wider existence in the early
1980s when they were picked up by the US-based Rodale Institute. Rodale, R
(1983) defined Regenerative Agriculture as ‘one that, at increasing levels of
productivity, increases our land and soil biological production base. It has a
high level of built-in economic and biological stability’. It provides for the
productive contribution of increasingly large numbers of people during a
transition to minimal reliance on non-renewable resources.
Philosophy of regenerative agriculture as presented by Harwood (1983)
are described as under

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 • Agriculture should produce highly nutritional food with high yield
and that should not be contaminated with biocides.
• Agriculture should increase the soil productivity in spite of decline,
by increasing the depth, fertility and physical characteristics of the
upper soil layers.
• It should maintain the nutrient-cycling by maintaining the flora and
fauna of soil.
• Crop production should rely on biological interactions for stability,
eliminating the need for synthetic pesticides.
• Agricultural production should generate increased levels of
employment.
• The integrated system should be utilized that will maintain the
ecosystem balance and give better crop production.
• Substances that leads to disruption in biological structure of the
farming system should not be used.
RA is soil centralized instead of self-centred and is based on the premise
that “health of soil, plants, animals, and humans is one and indivisible”
(Howard 1943; Howard and Howard 1945) from rattan lal. The aim of RA is
to apply the concept of more from less (McAfee 2019) to agriculture and
produce more from less (Lal 2013): less land area, less input of chemicals, less
use of water, less emission of greenhouse gases, less risk of soil degradation,
and less use of energy-based inputs (Table 1). Increasing availability of green
water by conserving precipitation, reducing losses by runoff and evaporation,
moderating soil temperature, and encouraging deep root systems is one of the
premises of regenerative agriculture. It involves the practices that help in
reduction/prevention of wind and water erosion so as to improve the soil
health and get better output from the farm. The reduction in hard pan,
compaction by using cover crop, residue retention are important practices
involved in regenerative agriculture.
Table 1: Potentials and aims of regenerative agriculture (Source: Lal, R 2020).

Parameter Expectations and outputs

Agronomic yield and
Optimum and sustainable
productivity
Inputs of chemicals Supplemental, as and when needed
Resource use Produce more per unit of land, water and energy
Global warming Positive soil/ecosystem carbon budget in accord with 4 per

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 1000 initiative, resilience to drought/heat waves and extreme
events, minimal emissions of methane and nitrous oxide
Profitability Optimal and sustained over time
Soil degradation, land
Reversed, and focused on degradation neutrality
desertification
Food quality Nutrition sensitive agriculture
Making farming integral to restoring and enhancing the
Environmental quality
environment
Legislation Soil quality act to complement the clean air & water act

Conclusion
There are tremendous benefits of Precision and regenerative agriculture,
but we cannot ignore the limitations associated with these technologies
because they create the hindrance for its adoption in the practical field
situation. Small land holdings, heterogeneity of cropping systems, high cost
technology, lack of local technical expertise and knowledge like computer
analysis and decision making are probably greater constraint in its success
path because technological gaps seen in the farmer community is making them
reluctant to rely on precision agriculture technology. Developments are going
on for rapidly mapping insect infestations, disease spread pattern study via
GPS and GIS receivers, variable rate spray operators are able to provide a
permanent record of where and when the treatment took place back to the field
manager by GPS data logger. Before the application of PA management, the
degree of variation needs to be assessed and the return on varying inputs
established. The justification for managing variation will depend on the
attribute, the magnitude of the variation, and cost-to-benefit ratio compared to
a uniform treatment. The use of precision agriculture is highly correlated with
regenerative agriculture and a much better step towards sustainability of the
ecosystem and crop production.
References
1. Ajai, (2002). GPS and its applications, Training course on Remote
Sensing and GIS Applications in Agriculture, May 27th -7th June, 2002,
RRSSC- Bangalore, pp: 89-99.
2. Bongiovanni, R., and Lowenberg-DeBoer, J. (2004). Precision
agriculture and sustainability. Precision agriculture, 5(4), 359-387.
3. Chang, K.T. (2008). Introduction to geographic information
systems (Vol. 4). Boston: McGraw-Hill.
4. Daponte, P., De Vito, L., Glielmo, L., Iannelli, L., Liuzza, D., Picariello,
F., and Silano, G. (2019). A review on the use of drones for precision

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 agriculture. In IOP conference series: earth and environmental science,
275 (1): 012022.
5. Duncan, J., Carolan, M. S., & Wiskerke, J. S. (Eds.), (2021). Routledge
handbook of sustainable and regenerative food systems. London, UK:
Routledge.
6. Harwood, RR (1983). International overview of regenerative agriculture.
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Tanzania, Morogoro, Tanzania, 16-20 May 1983, Faculty of Agriculture,
Forestry, and Veterinary Science, University of Dares Salaam, Morogoro,
TZ: Rodale Press
7. Howard, A. 1943. An agricultural testament. Nature 147(3733):590-591.
8. Howard, A., and L.E. Howard, (1945). Farming or Gardening for Health
or Disease (Published later as The Soil Health: A Study of Organic
Agriculture, 2010.) London, UK: Faber and Faber.
9. Jensen, J.R. (2009). Remote sensing of the environment: An earth
resource perspective 2/e. Pearson Education India.
10. Kurtza, D. B., Schellberg, J., and Braun, M. (2010). Ground and satellite-
based assessment of rangeland management in sub-tropical
Argentinaq. Applied Geography, 30: 210-220.
11. Lal, R. (2013). Food security in a changing climate. Ecohydrology &
Hydrobiology, 13(1): 8-21
12. Lal, R. (2020). Regenerative agriculture for food and climate. Journal of
soil and water conservation, 75(5): 123A-124A.
13. Lowenberg‐DeBoer, J., and Swinton, S. M. (1997). Economics of site‐
specific management in agronomic crops. The state of site-specific
management for agriculture, 369-396.
14. McAfee, A. (2019). More from Less: The Surprising Story of How We
Learned to Prosper Using Fewer Resources-And What Happens Next.
New York: Scribner
15. Oza, S. R., Panigrahy, S., & Parihar, J. S. (2008). Concurrent use of active
and passive microwave remote sensing data for monitoring of rice
crop. International Journal of Applied Earth Observation and
Geoinformation, 10(3): 296-304.
16. Rodale, R. (1983). Breaking new ground: The search for a sustainable
agriculture. Futurist, 17(1): 15-20.

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17. Seelan, S. K., Laguette, S., Casady, G. M., and Seielstad, G. A. (2003).
Remote sensing applications for precision agriculture: A learning
community approach. Remote sensing of environment, 88(1-2): 157-169
18. Shanwad, U. K., Patil, V. C., Dasog, G. S., Mansur, C. P., and Shashidhar,
K. C. (2002, October). Global positioning system (GPS) in precision
agriculture. In Proceedings of Asian GPS conference (Vol. 1)
19. Venkataratnam, L. (2002). Remote sensing and GIS in agricultural
resources management. In Proceedings of the First National Conference
on Agri-Informatics (NCAI), Dharwad, India, 3-4 June 2001 (pp. 20-29).
[Link]
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Measuring performance in precision agriculture: CART—A decision tree
approach. Agricultural water management, 84(1-2), 173-185.
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Remote Sensing. In IOP Conference Series: Materials Science and
Engineering, 490(6): 062014. IOP Publishing.

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Page | 290
 Chapter - 19
Future of Nanotechnology in Agriculture

Authors
Sonal Kumari
Department of Soil Science and Agricultural University,
Institute of Agricultural Science, Banaras Hindu University,
Uttar Pradesh, India
Ved Prakash
Department of Agronomy, Institute of Agricultural Science,
Banaras Hindu University, Uttar Pradesh, India

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Page | 292
 Chapter - 19
Future of Nanotechnology in Agriculture
Sonal Kumari and Ved Prakash

Abstract
Numerous scientists have really been drawn to nanotechnology because
of its diverse physical, chemical, and biological features, therefore, refers to
the creation and utilization of materials, devices, and systems through the
control of their properties and structure at a nanometric scale. Nanomaterials
had demonstrated numerous significant advantages for agricultural
applications including the ability to manipulate crop genetics and develop
nano-fertilizers and nano-complexes, nano-biosensors, nano-pesticides, and
nano-herbicides. Nanotechnology has allowed for the detection and
treats plant diseases, as well as enabled plants to absorb nutrients and
photosynthesize more quickly. Active substances of nano-fertilizer can also
be delivered to precise locations, and water can be treated using a variety of
nanomaterials. Nano-sensors also give accurate soil moisture updates at the
root zone and soil temperature. Nano-pesticides are three-fold more efficient
than conventional pesticides in controlling pests. Ultimately with proper and
controlled use of nanotechnology, it can support sustainable agriculture by
using optimum resources and ultimately enhancing crop productivity in near
future.
Keywords: Nanotechnology, nanomaterials, nano-fertilizers, nano-
herbicides, nano-pesticides
1. Introduction
“Nanotechnology is the art and science of manipulating matter at the
nanoscale.” Nanotechnology is the manipulation or self-assembly of
individual atoms, molecules, or molecular clusters into structures to create
materials and devices with new and vastly different properties. “Nano” refers
to a size scale between 1 to 100 nm at least in one dimension (NNI, 2005).
Agriculture is the backbone of most developing countries, with more than
60% of the population reliant on it for their livelihood. Agricultural scientists
are facing a wide spectrum of challenges such as stagnation in crop yields, low

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nutrient use efficiency, declining soil organic matter, multi-nutrient
deficiencies, climate change, shrinking arable land and water availability and
shortage of labour besides the exodus of people from farming. Despite the
immense constraints faced, we need to attain sustainable growth in agriculture
at the rate of 4% to meet the food security challenges. To address these
problems, there is a need to explore one of the frontier technologies such as
‘Nanotechnology’ to precisely detect and deliver the correct quantity of
nutrients and pesticides that promote productivity while ensuring
environmental safety and higher use efficiency. Nanotechnology can be
exploited in the value chain of the entire agriculture production system
(Subramanian and Tarafdar, 2011). Nanotechnology is emerging as the sixth
revolutionary technology in the current era after the Industrial Revolution of
the mid-1700s, the nuclear energy revolution of the 1940s, the green
revolution of the 1960s, the information technology revolution of the 1980s
and the biotechnology revolution of the 1990s. It is now an emerging and fast-
growing field of science which is being exploited over a wide range of
disciplines such as physics, chemistry, biology, material science, electronics,
medicine, energy, environment and health sectors.
Nanotechnology deals with matter at nanoscale (1-100 nm) dimensions.
These materials when reduced to the nanoscale show some properties which
are different from what they exhibit on a macro scale, enabling unique
applications. Nanoscience has brought a revolution in different fields by
helping develop processes and products that are hardly possible to evolve
through conventional methods. The nanotechnology-aided applications have
the potential to change agricultural production by allowing better management
and conservation of inputs for plant and animal production. A survey by
Salamanca-Buentella et al., (2005) predicted several nanotechnology
applications for agricultural production in developing countries within the
next 10 years. These included -
i) Nanoforms zeolites for slow release and efficient dosage of water
and fertilizers for plants; drugs for livestock; nanocapsules and
herbicide delivery.
ii) Nanosensors for soil quality and for plant health monitoring;
nanosensors for pest detection.
iii) Nanomagnets for removal of soil contaminants.
iv) Nanoparticles for new pesticides, insecticides, and insect repellents.
The recent statistics suggest that about 90% of the nano-based patents and
products have come from just seven countries, namely, China, Germany,

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France, Japan, Switzerland, South Korea and the USA while India’s
investments and progress is far from satisfactory. However, to take advantage
of the fascinating field of nano-science, the Government of India has invested
Rs. 1000 crore through the Nano Mission Project during the 11th Five Year
Plan and the investment is likely to be several folds higher during the 12th Five
Year Plan period.
The Indian Council of Agricultural Research (ICAR) has opened up an
exclusive platform to target Nanotechnology Applications in Agriculture. The
ICAR - Nanotechnology Platform encompasses major themes such as the
synthesis of nano-particles for agricultural use, quick diagnostic kits for early
detection of pests and diseases, nano-pheromones for effective pest control,
nano agri-inputs for enhanced use efficiencies, precision water management,
stabilization of organic matter in the soil, nano food systems and biosafety
besides establishing the policy framework. Green synthesis and microbial
synthesis of nanomaterials for their agricultural use may be very important as
they are naturally encapsulated with mother protein, therefore, more stable and
safer for a biological system. At present in India research is mainly
concentrated on nanoparticle synthesis, the smart release of nutrients from
nano-fertilizers, nano-induced polysaccharide powder for moisture
retention/soil aggregation and C build-up, regulated release of active
ingredients from nano-encapsulated herbicides, nano-seed invigoration, and
slow and steady release of pesticides, nano-film for extended shelf-life of
perishables and nano-remediation of soil and aquatic pollutants. These are
cutting-edge researchable areas which are expected to expand in the years to
come. However, if the nano products and the processes for creating them are
not managed judiciously, there could be a serious health and environmental
risks.
Background of nanotechnology

2000 years
Sulfide nanocrystals were used by Greeks and Romans to dye hairs
ago
1000 Years Gold nanoparticles of different sizes are used to produce different
ago, colours in stained glass windows
1959 “There is plenty of room at the bottom” by R. Feynman
1974 Taniguchi uses the term nanotechnology for the first time
1981 IBM develops Scanning Tunneling Microscope
“Buckyball” - Scientists at Rice University and the University of
1985 Sussex discover C60
“Engines of Creation” - First book on nanotechnology by K. Eric
1986
Drexler.

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 Atomic Force Microscope invented by Binnig, Quate and Gerbe
1989 IBM logo made with individual atoms
1991 Carbon nanotube discovered by S. Iijima
st
1999 “Nanomedicine” - 1 nanomedicine book by R. Freitas
2000 “National Nanotechnology Initiative” launched

Global scenario
The global nanotechnology market had touched, approximately US$ 29
billion by 2010 (Subramanian and Tarafdar, 2011). The exponential growth of
global investments in nanotechnology research can directly correspond with
the number of patents filed related to technology and products developed from
nanotechnology and nanoscience. Global market forecasts indicate USD1880
billion in investments in nanotechnology industries by 2015.
Nanoparticles
• Arranged or assembled into ordered layers, or mine layers.
• Revolutionize agriculture including pest management shortly.
• Possess distinct physical, biological and chemical properties
associated with their atomic strength.
Properties of nanoparticles
• Below about 100 nm the rules that govern the behaviour of the
elements of our known world start to give way to the rules of
quantum mechanics, and everything changes with Quantum effects.
Things behave differently in nano-world
• Carbon in the form of graphite (i.e. pencil lead) is soft, at the nano-
scale, can be stronger than steel and is six times lighter.
• Nano-scale copper is a highly elastic metal at room temperature,
stretching to 50 times its original length without breaking.
• Shiny orange yellow Gold changes its colour to brownish black on
reducing the size.
2. Methods of nanoparticle production
There are two processes in nanomaterial creation including "bottom-
up" processes (such as self-assembly) that create nanoscale materials from
atoms and molecules, and "top-down" processes (such as milling) that create
nanoscale materials from their macro-scale counterparts. Nanoscale materials
that have macro-scale counterparts frequently display different or enhanced

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properties compared to the macro-scale form (Petit et al., 1993). Such
engineered or manufactured nanomaterials will be referred to as “intentionally
produced nanomaterials,” or simply “nanomaterials.” The definition of
nanotechnology does not include unintentionally produced nanomaterials,
such as diesel exhaust particles or other friction or airborne combustion
byproducts, or nanosized materials that occur naturally in the environment,
such as viruses or volcanic ash (Roco, 2003). Information from incidentally
formed or natural nanosized materials (such as ultrafine particulate matter)
may aid in the understanding of intentionally produced nonmaterial.
Table 1: Following the development of complexity in nanotechnology (Tarafdar et
al., 2013).

Foremost infuse area of applications in

Generation Period Theme
which advancement happens
Up to Passive The top-down approach, nano-structured
First
2001 nanotechnology metal, polymer, ceramics, catalyst, MEMS.
The bottom-up approach, adaptive
2001- Active
Second nanostructure, solar cell transistors, sensor,
2005 nanotechnology
diagnostic assay, NEMS
Biomimetic nanostructure, novel
2005- Nanosystem
Third therapeutics, targeted drug delivery,
2010 technology
nanochips, agriculture.
2010- Molecular Atomic manipulations and design of
Fourth
2015 nanotechnology nanoscale architecture

3. Type of nanomaterials
3.1 Carbon-based nanomaterials
These nanomaterials are composed mostly of carbon, most commonly
taking the form of hollow spheres, ellipsoids, or tubes. Spherical and
ellipsoidal carbon nanomaterials are referred to as fullerenes, while cylindrical
ones are called nanotubes. These particles have many potential applications,
including improved films and coatings, stronger and lighter materials, and
applications in electronics (Oberdorster et al., 2006).
3.2 Metal-based nanomaterials
These nanomaterials include quantum dots, nanogold, nano silver and
metal oxides, such as titanium dioxide, zinc oxide, magnesium oxide, iron
oxide etc. A quantum dot is a closely packed semiconductor crystal comprised
of hundreds or thousands of atoms, and whose size is on the order of a few
nanometers to a few hundred nanometers. Changing the size of quantum dots
changes their optical properties (Dreizin, 2009).

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3.3 Dendrimer
These nanomaterials are nanosized polymers built from branched units.
The surface of a dendrimer has numerous chain ends, which can be tailored to
perform specific chemical functions. This property could also be useful for
catalysis. Also, because three-dimensional dendrimers contain interior
cavities into which other molecules could be placed, they may be useful for
drug delivery (Astruc et al., 2010).
3.4 Nanocomposites
Combine nanoparticles with other nanoparticles or with larger, bulk-type
materials. Nanoparticles, such as nanosized clays, are already being added to
products ranging from auto parts to packaging materials, to enhance
mechanical, thermal, barrier, and flame-retardant properties (Ajayan et al.,
2003).
Table 2: Beneficiary concentration (s) of nanoparticles for plants.

Beneficiary Part of
Nanoparticle (s) Plant Reference (s)
concentration (s) plant/process
Anjum et al.
Graphene oxide 400 and 800 mg/L Vicia faba L. Germination
(2014)
Germination
Lycopersicum Morla et al.
CNTs 40 µg/mL and seedling
esculantum (2011)
growth
Medicago
saliva, Root Miralles et al.
75 wt% CNTs
Triticum elongation (2012)
aestivum
Allium cepa,
9, 56, 315, and Root Cañas et al.
SWCNTs Cucumis
1,750 mg/L elongation (2008)
sativus
Hordeum
vulgare L., Lahiani et al.
MWCNTs 25-100 µg/mL Germination
Glycine max, (2013)
Zea mays
Lycopersicon Plant height
50 and 200 Khodakovskaya
esculentum and number of
µg/mL et al. (2013)
Mill flowers
5 up to 500 Nicotiana Khodakovskaya
Growth
µg/mL tabacum et al., (2012)
Root growth,
Triticum Wang et al.,
o-MWCNTs 10-160 µg/mL vegetative
aestivum (2012a)
biomass
wsCNTs 6.0 µg/mL Cicer Growth rate Tripathi et al.,

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 arietinum (2011)
Uptake
Lycopersicon
nutrients (K, Tiwari et al.,
MWCNTs, 40 µg/mL esculentum
Ca, Fe, Mn (2013)
Mill
and Zn)
Nutrient
Pristine Tiwari et al.,
20 mg/L Zea Mays transport,
MWCNTs (2014)
biomass
Micronutrients
Cucumis Zhao et al.,
ZnO NPs 400 mg/kg (Cu, Mn and
sativus fruit (2014)
Zn)
1.5 ppm (foliar Cicer Shoot dry Burman et al.,
spray) arietinum L weight (2013)
20 ppm
Dhoke et al.,
(suspension, foliar Vigna radiata Biomass
(2013)
spray)
Arachis Prasad et al.
1,000 ppm Germination
hypogaea (2012)
Stem, root
Arachis Prasad et al.,
1,000 ppm growth and
hypogaea (2012)
Yield
500, 1,000 and Vigna radiate Patra et al.,
Dry weight
2,000 L. Wilczek (2013)

10 and 80 Arabidopsis
GNPs Germination Kumar et al., (2013)
µg/mL thaliana
10 and 80 Arabidopsis
Root length Kumar et al., (2013)
µg/mL thaliana
Shoot and root
Arabidopsis
10 µg/mL system(longer), early Kumar et al., (2013)
thaliana
flowering, yield
Boswellia Germination and Savithramma et al.,
AgNPs 10-30 µg/mL
ovaliofoliolata seedling growth (2012)
Phaseolus
60 ppm vulgaris L., Root length Salama (2012)
Zea mays L.
Phaseolus
60 ppm vulgaris L., Shoot length Salama (2012)
Zea mays L.
Phaseolus
Dry weight of root
60 ppm vulgaris L., Salama (2012)
and shoot
Zea mays L.
Antagonize inhibition
Karuppanapandian et
100 µM Vigna radiata by 2,4- D at 500 μM
al. (2011)
of plant growth

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 500, 1,000,
Sulfur
2,000 and Vigna radiata Dry weight Patra et al., (2013)
NPs
4,000 ppm
Yuvakkumar et al.,
SiO2NPs 15 kg/ha Zea mays L. Growth parameters
(2011),
TiO2 Arabidopsis
400 mg/L Root length Lee et al., (2010)
NPs thaliana,
Foenicutum
60 ppm Germination Feizi et al., (2013)
vulgare
lower than
Lemna minor Plant growth Song et al., (2012)
200 mg/L
Triticum Mahmoodzadeh et
1,000 mg/L Chlorophyll content
aestivum al., (2013)
Hill reaction, non-
cyclic
Spinacia Hong et al., (2005a,
0.25 % photophosphorylation,
oleracea b)
protect chloroplasts
from aging
Net photosynthetic
Lycopersicon rate, conductance to
0.05-0.2 g/L esculentum H2O, and transpiration
Qi et al., (2013)
Mill rate, Regulation of
photosystem II (PSII)

0.25%
Nano-anatase Spinacia Rubisco activase (rca) Ma et al.,
(foliar
TiO2 oleracea mRNA expressions, (2008)
spray)
Oxygen evolution,
0.25% Rubisco carboxylation,
Spinacia Gao et al.,
(foliar Rubisco Activase, rate
oleracea (2006)
spray) of photosynthetic
carbon reaction
Spinacia Several enzymes Yang et al.,
0.25%
oleracea activities induction (2006)
Aluminum oxide 400-4,000 Arabidopsis Lee et al.,
Root length
NPs mg/L thaliana, (2010)
Juhel et al.,
Alumina NPs 10 mg/L Lemna minor Root length
(2011)
Juhel et al.,
0.3 g/L Lemna minor Biomass accumulation
(2011)
nZVI Zero-Valent
Iron (nanoscale Arabidopsis Kim et al.,
0.5 g/L Root elongation
particles) Iron thaliana (2014)
oxide NPs
0.5-0.75 Glycine max Yield and quality Sheykhbaglou
Iron oxide NPs
g/L50 ppm Vigna Biomass et al., (2010)

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 (foliar radiata
spray)
50 ppm
ZnFeCu-oxide Vigna Dhoke et al.,
(foliar Biomass
NPs (suspension) radiata (2013)
spray)
Arabidopsis Ma et al.,
CeO2 NPs 250 ppm Biomass
thaliana (2013)
Raphanus Wu et al.,
CO3O4 NPs 5 g/L Root elongation
sativus L. (2012)
500 mg/kg
Triticum Dimkpa et al.,
CuO NPs (sand Biomass
aestivum (2012)
culture)
Hydroxyapatite 100-2,000 Lactuca Wang et al.,
Root length
suspension mg/L sativa (2012b)

Why should agriculture scientists think of nanotechnology?

1) Crop yield stagnation
2) Declining organic matter
3) Multinutrient deficiencies
4) Climate change
5) Shrinking arable land and water availability
6) Resistance to GMOs crops
7) Shortage of labour
4. Application of nanotechnology in agriculture and allied science
Agriculture is the backbone of most developing countries, with more than
60% of the population reliant on it for their livelihood (Brock et al., 2011). As
well as developing improved systems for monitoring environmental
conditions and delivering nutrients or pesticides as appropriate,
nanotechnology can improve our understanding of the biology of different
crops and thus potentially enhance yields or nutritional values. In addition, it
can offer routes to added-value crops or environmental remediation.
Nanotechnology, as a new enabling technology, has the potential to
revolutionize agriculture and food systems. Agricultural and food systems
security, disease treatment delivery systems, new tools for molecular and
cellular biology, and new materials for pathogen detection and protection of
the environment are examples of the important links of nanotechnology to the
science and engineering of agriculture and food systems (Welch and Graham,
1999). Nanotechnology operates at the same scale as a virus or disease-
infecting particle, and thus holds the potential for very early detection and

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eradication. Nanotechnology holds out the possibility that “Smart” treatment
delivery systems could be activated long before macro symptoms appear. For
example, a smart treatment delivery system could be a miniature device
implanted in an animal that samples saliva regularly. Long before a fever
develops, the integrated sensing, monitoring and controlling system could
detect the presence of disease and notify the farmer and activate bioactive
systems such as drugs, pesticides, nutrients, probiotics, nutraceuticals and
implantable cell bioreactors.

Fig 1: Application of nanotechnology in agriculture (Shang et al., 2023)

4.1 Nanotechnology on food
The term “nanofood” may be defined as Food that has been cultivated,
produced, processed, or packed using nanotechnology tools or techniques, or
to which artificial nanoparticles have been introduced, is now referred to as
"Nanofood" (Meghani et al., 2020). It does not mean atomically modified food
or food produced by nanomachines. Although there are ambitious thoughts of
creating molecular food using nanomachines, this is unrealistic in the
foreseeable future. Instead, nanotechnologists are more optimistic about the
potential to change the existing system of food processing and to ensure the
safety of food products, creating a healthy food culture. They are also hopeful

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of enhancing the nutritional quality of food through selected additives and
improvements to the way the body digests and absorbs food. Although some
of these goals are further away, the food packaging industry already
incorporates nanotechnology in products. In Agriculture, nanotechnology is
used in the entire field to enhance productivity by using smart delivery
systems of fertilizer, insecticide, herbicide and growth regulators. The related
fields are as under
4.1.1 Food science and technology
Nanotechnology has been used to create nanosensors for the quick and
sensitive detection of potentially harmful ingredients in food as well as a smart
packaging system. The most important technology in food science is
nanoencapsulation, particularly for bioactive ingredients and flavours. After
oral administration, targeted delivery systems using nanoencapsulation can
improve the bioavailability of bioactive substances. Additionally,
nanoencapsulation makes it possible to regulate the timing of flavour release
and guard against flavour deterioration during processing and storage (Yu et
al., 2018). Flavours are regarded as essential components in all foods since
they have a big impact on sensory quality and how much people eat. The
quality and acceptance of the food are related to the growing interest in the
stability of flavours in many forms of food. However, controlling and
stabilizing tastes can be challenging, particularly during the manufacturing
and storage procedures. It is advantageous to encapsulate the flavour before
using it in food, boosting chemical stability and offering controlled release, to
reduce flavour degradation or loss during processing and storage. A protective
carrier that is encapsulated prevents oxidation, light-induced reactions, and
taste interactions. Biopolymers such carbohydrates (such as starch,
maltodextrins, and dextrose), gums (such as gum arabic, alginates, and
carrageenan), proteins (such as whey proteins and gelatin), and chitosan are
frequently used as transporters. (Chang et. al., 2019). Nanotechnology for
enhancing food security in India (Sastry et al., 2011). Hybrid polymer: Smart
packaging with Nano silicon-embedded durethan polymer to enhance the shelf
life of the food materials Bayer polymers.
4.2 Crop improvement
The capacity to sequence DNA in nanofabricated gel-free systems, which
would enable substantially faster DNA sequencing, is being researched in
nanobiotechnology. DNA sequencing data of the crop germplasm, including
the cultivated crop gene pool and the wild relatives, when combined with
potent methods like association genetic analysis, has the potential to yield

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highly valuable information about molecular markers associated with
economically and agronomically significant traits. Thus, molecular marker-
assisted breeding for crop improvement can advance more quickly thanks to
nanobiotechnology (Verma et al., 2021). Gene therapy for plants: use of 3-nm
mesoporous silica nanoparticle (MSN) for smuggling foreign DNA into cells.
(Peterson et al.,2014). Shown the feasibility of DNA sequencing using a
fluidic nanochannel functionalized with a graphene nanoribbon (kyu min et
al., 2013).
4.3 Seed technology
Seed is the most important input determining the productivity of any
crop. Conventionally, seeds are tested for germination and distributed to
farmers for sowing. Although seed testing is done in well-equipped
laboratories, it is hardly reproduced in the field due to inadequate moisture
under rainfed conditions. In India, more than 60% of the net area sown is
rainfed; hence, developing technologies for rainfed agriculture is quite
appropriate. A group of research workers is currently working on metal oxide
nanoparticles and carbon nanotubes to improve the germination of rainfed
crops. Khodakovskaya et al., (2009) have reported the use of carbon
nanotubes for improving the germination of tomato seeds through better
permeation of moisture. Their data show that carbon nanotubes (CNTs) serve
as new pores for water permeation by penetration of the seed coat and act as a
passage to channel the water from the substrate into the seeds. These processes
facilitate germination which can be exploited in rainfed agricultural systems.
The use of carbon nanotubes increases germination through better penetration
of moisture.
The effective process of seed nano-priming can alter the metabolic and
signalling pathways of seeds, which has an impact on all stages of plant life,
including germination and seedling establishment. Studies have demonstrated
several advantages of seed nano-priming, including enhanced plant growth
and development, increased productivity, and better food quality in terms of
nutrients. Nano-priming modulates biochemical pathways and the balance
between reactive oxygen species and plant growth hormones, resulting in the
promotion of stress and disease resistance outcoming in the reduction of
pesticides and fertilizers (Do et al., 2021).
4.4 Precision farming
Precision farming offers significant possibilities for using
nanotechnology. It is simple to create nanoparticles with distinctive features
from many biological sources, and these particles can be used in agriculture.

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Plant extracts (leaves, flowers, stems, and roots) from a variety of plant species
have been effectively exploited as biological sources to create nanoparticles.
Metal ions are converted to nanoparticles in one step by biomolecules found
in plant extracts. This green synthesis process is environmentally benign,
simple to carry out, can be done at room temperature without the use of
complicated equipment, and is simple to scale up or modify to meet different
needs. Alkaloids, phenolic compounds, terpenoids, and other water-soluble
plant metabolites as well as co-enzymes are reduced to nanoparticles in this
process. With the use of controlled-release formulations of nanoparticles, the
nanotechnology-based distribution of nano-particles has shown promising
outcomes for plant disease resistance, increased plant development, and
nutrition via site-specific delivery of fertilizers and other critical nutrients (Yu
et al., 2018).
4.5 Smart field system
Smart field sensing systems are increasingly important applications for
the real-time monitoring of crop growth and field conditions including
nutritional status, light, temperature, moisture level, soil fertility, insects,
weeds and plant diseases to maximise yields for sustainability in changing
climatic conditions and to maximise yield to feed a growing world population
(Chen and Yada, 2011). Bio-Nanotechnology has created sensors that are
more sensitive, quicker to react to environmental changes, and connected to
GPS. These cover large regions and keep an eye on crop growth and soil
conditions. Such sensors have already been used in UASD Plant pathology in
the US and Australia. At 10 ppm of the nanosized silica-silver13, Rhizoctonia
solani, Botrytis cinere, Magnaporthe grisea, Colletotrichum gloeosporioides,
and Pythium ultimum all displayed 100% growth inhibition. QDs have become
an essential instrument for the highly accurate identification of a specific
biological signature. (Madhuri et al., 2010).
4.6 Soil remediation
Nanotechnology's use in soil remediation to get rid of toxins in the soil
environment has become a major global problem. The fate of contaminants in
soil environments, the mechanisms of nanotechnology with various types of
nanomaterials for soil remediation, the benefits and drawbacks of
nanomaterials for terrestrial organisms, human health, and the soil
environment, as well as the difficulties of applying nanotechnology for soil
remediation were all covered in this review. The detrimental impact of
nanoparticles on bacteria is one of nanotechnology's main problems.
Nanomaterial toxicity will have an impact on microorganisms and prevent
enzyme activity in the soil environment (Kristanti, et al., 2021).

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4.7 Removal of heavy metals
In recent decades, the amount of heavy metal water contamination has
significantly increased. Therefore, it is necessary to create strategies,
regulations, technologies, and materials to address this issue and prevent its
potentially disastrous impacts on the environment and human health. One
practical method for addressing heavy metal water pollution is
nanotechnology. Engineered nanoparticles have special physicochemical
characteristics that make them effective at removing hazardous metal ions
from contaminated water. Different types of nanoparticle-structured materials,
including carbonaceous, zeolite, polymer-based, magnetic and metal oxide,
carbon materials, and magnetic and silica-based materials, demonstrated a
high removal efficiency for various types of heavy metals with excellent
adsorption capacity and selectivity even at very low concentrations.
Temperature variation, pH change, adsorbent dose, starting concentration, and
ionic strength are major factors impacting the adsorption process. Toxic metal
ions can be quickly removed and adsorption equilibrium can be reached by
optimizing these parameters. There are several drawbacks associated with the
usage of sorbents; occasionally, they congregate and are unstable, which
reduces the effectiveness of their removal. Because of their small size, they
need to be effectively detached from metal ions. Given that heavy metals have
a strong absorption tendency, ligand-based nanocoating can be used to remove
them effectively (Ethaib. et al., 2022).
4.8 Nano-based water management
For the treatment of surface water, groundwater, and wastewater
contaminated by harmful metal ions, organic and inorganic solutes, and
microbes, nanotechnology holds the promise of innovative nanomaterials.
Many nanomaterials are being researched and developed for application in
water purification due to their distinct action towards resistant pollutants.
Pathogens in water need to be quickly and accurately identified to ensure
public health. Traditional laboratory testing, however, takes a lot of time.
Enzyme-based, immunological, or genetic test-based faster procedures are
being developed (Elizabeth et al., 2019). Nanofiber membranes and nano
biocides, which seem to be promisingly successful, may help with water
filtering. Mats of bacteria encased in natural polymers, known as biofilms,
contaminate drinkable water and are challenging to remove with
antimicrobials or other chemicals. They can only be cleaned up mechanically,
which requires a lot of labour and downtime. Enzyme therapies that could be
able to dissolve such biofilms are being developed. Arsenic removal from
tainted water using magnetite (iron oxide) nanocrystals. Market-available

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water purifiers use a variety of methods, including distillation, filtration,
boiling, chlorination, sedimentation, and oxidation. Currently,
nanotechnology is essential to water filtration methods. Nanoscale atom
manipulation is the practice of nanotechnology. Nanomembranes are used in
nanotechnology to soften water and remove impurities like chemical,
biological, and physical pollutants. Numerous methods in nanotechnology use
nanoparticles to effectively and safely provide safe drinking water. Some
methods have been used by the industry. Nanotechnology is chosen for
improved water purification or treatment procedures. Processes for treating
water use a wide variety of nanomaterials and nanoparticles. Regarding
cleanup, desalination, filtration, purification, and water treatment,
nanotechnology is helpful.
4.9 Nano-fertilizer technology
Fertilizers, together with good seed and irrigation, were mostly
responsible for India's increased food grain production, which increased from
55 mt in the 1960s to 254 mt in 2011 and coincided with a dramatic rise in
fertilizer consumption from 0.5 mt to 23 mt. It has been unequivocally proven
that fertilizer accounts for between 35 and 40 per cent of any crop's
productivity. Given its significance, the Indian government extensively
subsidises the price of fertilizers, notably urea. As a result, fertilization has
become unbalanced, and in certain places, excessive nitrogen application has
led to groundwater nitrate pollution. In the past few decades, the utilization
efficiency of N, P and K fertilizers have remained stable around 30-35%, 18-
20% and 35-40%, respectively, leaving a substantial amount of additional
fertilizers to build in the soil or enter into the aquatic system causing
eutrophication. It is critical to develop a nano-based fertilizer formulation with
multiple functions to address issues with low fertilizer use efficiency,
unbalanced fertilization, multi-nutrient deficiencies, and decline of soil
organic matter.
Although incredibly revolutionary, nano-fertilizer technology has not
been extensively covered in the literature. But several data and patents
strongly imply that there is a huge window of opportunity for the creation of
nano-fertilizers. Nanoparticle fertilizer applied topically has been found to
significantly enhance yields (Tarafdar, 2012; Tarafdar et al., 2012a). In an arid
climate, it was demonstrated that 640 mg ha-1 of foliar nano phosphors (at a
40-ppm concentration) produced an 80 kg ha-1 P equivalent yield of cluster
bean and pearl millet. The development of nano-composites to deliver all the
necessary critical nutrients in the right amounts via an intelligent delivery
system is currently being researched. Initial findings indicate that

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nanotechnology may be used to provide balanced fertilization (Tarafdar et al.,
2012b). In fact, it is necessary to determine the metabolic uptake of metals,
such as micronutrients, delivered as Nano-formulations by soil-borne and
foliar application or in other ways, inside the plant biomass. Furthermore, it's
important to carefully consider the Nano-composites being considered to
provide all the nutrients in the correct amounts through "Smart" delivery
methods. Due to the loss of 50-70% of the nitrogen provided by conventional
fertilizers, the nitrogen usage efficiency is currently low. By boosting plant
uptake, new nutrient delivery techniques that take use of plants' porosity
nanoscale structures could decrease nitrogen loss. The uptake of nutrients will
increase when fertilizers are contained in nanoparticles (Tarafdar et al.,
2012c). The release of the nutrients in the subsequent generation of nano
fertilizers may be prompted by an environmental factor or may simply occur
at a predetermined period. Cluster bean and pearl millet yielded equally after
receiving foliar fertilizer applications of nano phosphors (640 mg ha-1) and
phosphorous (80 kg ha-1) in an arid environment (Tarafdar et al., 2012).
Fertilizers play a pivotal role in agriculture production up to 35 to 40% of the
productivity. To enhance nutrient use efficiency and overcome the chronic
problem of eutrophication, nano fertilizer might be the best alternative.
Attempts have been made to synthesize nano fertilizer to regulate the release
of nutrients depending on the requirements of the crops, and it is also reported
that nano fertilizers are more efficient than ordinary fertilizers (Liu et al.,
2006).
4.10 Nanotechnology-based weed management
The threat of weeds in agriculture. Weeds may endanger the entire harvest
in the fragile agroecosystems because two-thirds of Indian agriculture is
rainfed cultivation, which uses relatively little herbicide. Market-available
herbicides are made to kill or control the weed plants' above-ground growth.
None of the herbicides prevents the growth of rhizomes or tubers, which are
active belowground plant portions that serve as a seed for new weeds the
following season. Lower yields than soils where weeds are controlled are
likely to result from weed infestations and weed seeds. Crop productivity may
increase if nanotechnology is used to increase the effectiveness of herbicides.
The encapsulated nano-herbicides are pertinent in light of the requirement to
develop and construct a nano-herbicide that is protected by the natural
environment and functions only during a period of rainfall, which accurately
mimics the rainfed system. The development of a target-specific herbicide
molecule enclosed in a nanoparticle is directed at a particular receptor in the
roots of the target weeds; this receptor allows the herbicide to enter the root

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system and translocate to areas that inhibit the glycolysis of food reserves in
the root system. This will cause the particular weed plant to run out of food
and be killed. (Chinnamuthu and Kokiladevi, 2007). Presently accessible
adjuvants for herbicide application purport to contain nanoparticles. When
used with a "nanotechnology-derived surfactant," one nano surfactant based
on soybean micelles has been found to make crops that are resistant to
glyphosate sensitive to it. According to reports, a soy-based nano surfactant
makes glyphosate-resistant crops sensitive to it (Kokiladevi et al., 2007).
4.11 Nanotechnology in pesticides
Pest populations can be reduced to the point at which management is no
longer effective by continuing to use pesticides during the early stages of crop
growth. Therefore, one of the most practical and economical ways to manage
insect pests is to utilise active substances on the surface that is being treated.
A nanotechnology strategy called "nano-encapsulation" can be utilised to
increase the insecticidal value by shielding the active ingredient from harmful
environmental factors and fostering persistence. Nanoencapsulation involves
sealing tiny amounts of the active substances inside a sac or shell with a thin
wall (protective coating). Insecticide encapsulation has been the subject of
several recent research publications. Insecticides, fungicides, and nematicides
can be nano-encapsulated to create formulations that effectively control pests
while limiting the buildup of residues in the soil. A nanotechnology approach
of "controlled release of the active ingredient" may be used to improve the
efficacy of the formulation, which may significantly reduce the amount of
pesticide input and associated environmental risks. This approach aims to
protect the active ingredient from degradation and to increase persistence.
Because the amount of product that is actually effective is at least 10-15 times
lower than that administered with conventional formulations, nano-pesticides
will minimise the rate of administration. As a result, a significantly smaller
amount may be needed to achieve much better and extended management.
Many pesticide producers are working on producing pesticides that are
enclosed in nanoparticles (OECD and Allianz, 2008). These insecticides may
release gradually over time or immediately in response to an environmental
trigger (for example, temperature, humidity, or light). It is uncertain if these
pesticide products will be readily available on the market shortly. Crop
production is significantly impacted by plant diseases. The identification of
the precise stage of prevention is where the disease management challenge
lies. The majority of the time, the crop is treated with the proper plant
protection chemicals as a preventative step, which can result in avoidable
environmental dangers. Alternatively, treatments are carried out once disease

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signs start to manifest, which can result in some crop losses. Viral infections
are among the most challenging to manage because they must be stopped
before they spread through their vectors. However, applying pesticides won't
be very effective after it starts exhibiting its symptoms. Therefore, the key to
successfully controlling viral infections is the precise stage detection, such as
the stage of viral DNA replication or the generation of early viral protein.
Nano-based viral diagnostics, including the creation of multiplexed
diagnostics kits, have gained traction to identify the precise virus strain and
the stage at which a particular treatment is being used to halt the sickness. An
additional developing field of research in bio-Nanotechnology is the
identification and application of biomarkers that reliably identify disease
stages. The identification of various proteins that are produced throughout the
infection cycle is made possible by measuring differential protein production
in both healthy and sick states. Clay nanotubes (halloysite) have been
developed as carriers of pesticides at low cost, for extended-release and better
contact with plants, and they will reduce the number of pesticides by 70-80%,
thereby reducing the cost of pesticides with minimum impact on water
streams.
4.12 Nano-scale carriers
The effective distribution of fertilizers, insecticides, herbicides, plant
growth regulators, etc. can be accomplished by using nanoscale carriers.
Encapsulation, trapping, polymers, dendrimers, surface ionic and weak bond
attachments are only a few of the mechanisms that contribute to effective
delivery, better storage, and controlled release. These contribute to increased
resistance to environmental deterioration and, ultimately, need less
application, which lessens chemical runoff and resolves environmental issues.
These carriers may be constructed to allow plant roots to be fastened to the
organic matter and soil particles in the vicinity. This is only conceivable if we
understand the molecular and structural relationships between soil fractions,
targeted nanoscale structures, and these relationships. As and when they are
made, these developments will aid in decreasing the absorption of active
substances, hence lowering the number of inputs needed and the waste
generated.
4.13 Nutrients and contaminants detection through biosensors
Rapid, sensitive, and precise molecular detection of contaminants and
pathogens is necessary for environmental and soil health protection. The same
set of protocols has been used for evaluating soil fertility for the past 60 years,
however, they may be out of date in light of modern production systems and

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precision farming techniques. For in-field real-time monitoring of vast areas,
precise sensors are required for remote sensing, small portable devices, and in
situ detection. These tools can shorten the time needed for protracted
immunoassays and microbiological testing. These tools are used for a variety
of purposes, including the identification of pollutants in various bodies,
including water supplies, raw food materials, and food items. Because they
connect to specific biomolecules in a highly specific way, enzymes can
function as a sensing component. A pattern of reaction across many gas
sensors is employed by the electronic nose (E-nose) to recognize various
odours kinds. Similar to how the human nose might detect an odour, it can
recognize the odorant, calculate its concentration, and discover its
distinguishing characteristics. It mostly consists of nanoparticle-based gas
sensors like ZnO nanowires. With the passage of a certain gas, its resistance
modifies, causing a change in an electrical signal that creates the fingerprint
pattern for gas identification. High-performance capabilities are offered by
biosensors for application in identifying pollutants in food or environmental
media. At a modest price, they provide great specificity and sensitivity, quick
reaction, straightforward operation, and small size (Amine et al., 2006). The
direct enzyme inhibition sensors may be useful as a screening tool to identify
when a sample contains one or more contaminants, even though they currently
lack the analytical ability to distinguish between multiple toxic substances in
a sample (such as the simultaneous presence of a heavy metal and a pesticide).
These techniques can be used with single-use test strips (making them useful
to those in the field). According to Hu et al., (2010), a nanomagnetic particle
was used in an enzyme-linked immunosorbent assay (ELISA) test to detect
various residues of organophosphorus pesticides. According to the authors,
ELISA is more economical than analytical techniques that demand high
degrees of expertise and pricey laboratory equipment.
4.14 Nanotechonogy-based agricultural engineering issues
Numerous uses of nanotechnology exist in the world of agricultural
engineering. The application in machine structure and agricultural tools to
increase their resistance against wear and corrosion and ultraviolet rays;
producing strong mechanical components with the use of Nano-coating and
use of bio-sensors in smart machines for mechanical-chemical weed control;
production of Nano-cover for bearings to reduce friction. It is also important
to mention how nanotechnology is used to produce alternative fuels and
reduce environmental pollution. Tools used in agriculture can be nanocoated
to increase their resistance to wear and corrosion.

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4.15 Nanotechnology in animal sciences
Food products, veterinary care, prescription drugs, and domestic animal
immunizations are just a few of the problems that can be solved with the help
of nanotechnology. When employed at the nano level, specific drugs including
antibiotics, vaccinations, and probiotics are beneficial in treating infections,
nutritional deficiencies, and metabolic problems. The use of drugs at the
nanoscale allows for the removal of biological barriers for greater therapeutic
efficacy. The key benefits of using nanotechnology in drug application are an
appropriate time for drug release and self-regulatory capacities.
The bucky ball-shaped C-60 carbon particle is a spherical molecule with
a diameter of about 1 nm. Its nature makes it biocompatible and non-toxic to
living cells. Drugs and peptides that are water soluble can be delivered using
it as a carrier. Understanding particular drug behaviours in an animal's body
can be aided by nanotechnology. The nanoparticles are believed to be
employed as sensors to identify altered cell behaviour because they may
reportedly penetrate the skin through minor abrasions. The synthetic three-
dimensional macromolecules known as dendrimers have a central particle that
is encircled by branches to resemble a tree. Because they are biocompatible
and quickly removed from circulation through the kidney, they can be
conjugated with the target molecule, such as a medication. It was found that
dendrimer-methotrexate delivered in vivo reduced tumour size ten times more
than free methotrexate (Hong et al., 2007).
The usage of nano-magnets as a drug delivery mechanism is particularly
useful for treating malignant growths without causing any damage to nearby
tissues. To create a nanoparticle-based medication delivery system, many
proteins can be employed, including albumin, gelatin, gliadin, and legumin.
Racing horses' osteoarthritis-causing antigen was neutralised using inert
nanobeads. Less antibiotic is used and less antibiotic residue is left behind
when treating animal infections with nano-based antibiotics. Chromium
supplementation based on nanoparticles improves muscle tissue chromium
concentration while improving growth performance and body composition.
Due to low bioavailability, iron insufficiency is a prevalent issue in animals,
particularly during the early stages of life, gestation, and parasitic infestation.
The addition of ferric phosphoric nanoparticles can improve bioavailability.
Large amounts of chicken or goat meat are produced in laboratories using
nanotechnology while preserving the same nutritional content, flavour, and
texture without posing any risks (vegetarian meat). Even vegetarians can eat
it. It may end hunger by addressing the issue of food scarcity. It is commonly
known that designer eggs are produced using nanotechnology. It can yield

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eggs with the necessary antibodies, more nutrients, fewer yolks, and reduced
cholesterol. Additionally, nano-based sensors can aid in the early detection of
infections that affect eggs (Verma et al., 2012). The effectiveness of utilising
nutrients in the feed is improved by complexes made of nanomicelles,
nanoliposomes, and nanoemulsions.
4.16 Nanotechnology in fisheries and aquaculture
Nanotechnology can completely transform the fisheries and aquaculture
industries. Numerous mysteries surrounding fisheries nutrition, health, and
reproduction could be resolved with the help of nanotechnology techniques
such as nanomaterials, nanosensors, DNA nano vaccines, gene delivery, and
smart medicine delivery. By detecting germs in packaging, providing strong
flavour, high-quality colour, and safety, nanotechnology will aid the fish
processing industry in producing high-quality products. The 40 nm
lanthanum-based chemical NanoCheck removes phosphates from the water
and inhibits the growth of algae. Iron nanoparticle feeding resulted in a 30%
higher development rate in younger carp.
An abstract model for timing the discharge of ZnO nanoparticles to match
crop demand Specific root chemical signals are bound to a nano biosensor
contained in a thin polymer film (blue), which coats ZnO-fertilizer
nanoparticles (yellow and red, respectively) (dark grey). The release,
dissolution, plant uptake, and aggregation of ZnO NPs (white spheres) in the
soil solution of crop rhizospheres are caused by the selective signal-biosensor
binding process.
5. Health and environmental concerns
Researchers discovered that rats exposed to nanoparticles typically
experience particle settling in the brain and lungs, which resulted in markedly
elevated levels of biomarkers for inflammation and stress response. They also
discovered that nanoparticles can cause skin ageing through oxidative stress
in hairless mice. Nanofibres, or extremely small fibres, can cause lung damage
comparable to that caused by asbestos. The Royal Society report identified a
risk of Nanoparticles or Nano-tubes being released during disposal,
destruction, and recycling, and it advised manufacturers of products covered
by extended producer responsibility regimes, such as end-of-life regulations,
to publish procedures outlining how these materials will be managed to
minimize potential human and environmental exposure.” The Institute for
Food and Agricultural Standards has suggested that standards for
Nanotechnology research and development be integrated across consumer,
worker, and environmental standards to reflect the difficulties in ensuring

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responsible life cycle regulation. They also suggest that NGOs and other
citizen organizations take an active part in creating these standards.
Unfortunately, the same property that gives nanoparticles their enormous
value also has some negative impacts and can pose serious risks to the
environment, animals, people, and plants when used carelessly. The following
list of potential risks is provided:
1) When airborne, nanoparticles from pesticides, fertilizers, or other
formulations may settle on a plant's above-ground portions. They
might close stomata and form a thin, poisonous barrier layer on the
stigma that prevents pollen tubes from penetrating. The movement of
water, minerals, and photosynthate may also be hampered by their
entry into the vascular tissue.
2) Animals may inhale nanoparticles, which can cause a range of
diseases and problems. The particles could go into the blood.
3) Nano-pesticides may lessen environmental contamination by
reducing pesticide application rates, but because of their accelerated
transport, prolonged persistence, and increased toxicity, they may
also introduce new types of contamination into soils and streams.
4) Airborne nanoparticles can enter the body through the respiratory
system and pose some specific health risks to people. There is a
chance that inflammation, protein fibrillation, and the production of
genotoxicity will occur as a result of nanoparticles entering the
bloodstream and lungs.
The use of nanotechnology in fertilizers and pesticide formulations must
be approached extremely cautiously due to these concerns, which necessitates
the requirement for critical analyses and assess the dangers associated with the
nano-formulations.
6. Conclusion
Nanotechnology has the potential to be employed in agricultural products
that safeguard plants, track their progress and spot diseases. Researchers are
still looking for new ways to use nanotechnology in agriculture and the food
sector. In fact, there will be significant improvements to the agriculture and
food industries in the upcoming years.
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