Lecture 16.

11. BIOFERTILIZERS

In a broader sense, biofertilizers consist of both microbial biofertilizers and fertilizers

of biological origin such as compost and vermicompost. However, in general usage, the term
‘biofertilizer’ is restricted to refer to microbial biofertilizers only. A microbial biofertilizer is
a preparation containing live or latent cells of efficient strains of microorganisms applied to
seed, soil, or composting areas with the objective of increasing their population and
accelerating certain microbial processes to augment the extent of availability of nutrients in a
form that can be easily assimilated by plants.
NITROGEN BIOFERTILIZERS
Rhizobia
The bacteria collectively known as “rhizobia” consist of five genera of bacteria,
Rhizobium, Bradyrhizobium, Mesorhizobium, Sinorhizobium, and Azorhizobium. They6
induces better nodulation and stem nodulation in inoculated plants, and thereby, brings down
the requirement of nitrogen fertilizers for pulses, and leguminous oil seeds and green
manures. It is available as carrier based inoculum. Use Rhizobium cultures at the rate of 250-
375 g/ha. Mix inoculants of a packet with about 200 ml of cold rice water or 2.5 percent
starch solution, and to this add seeds required for 1 ha. Treat the seeds using your hands. Sow
the treated seeds within 24 hours.
In acidic soils, activity of Rhizobium is reduced. To overcome this problem, lime
pelleting of Rhizobium treated seeds is recommended. For 10 kg of seeds, use 0.5 to 1.5 kg
of finely powdered (300-mesh) calcium carbonate depending on the size of seeds. Add
calcium carbonate powder to moist freshly treated seeds, and mix for 1 to 3 minute until each
seed is uniformly pelleted. Spread out the pellets on a clean paper to harden. Although lime
pelleted seeds can be stored up to one week in a cool place, it is better to sow them as soon as
possible.
Azospirillum
Azospirillum is a free-living bacterium that grows on and inside the roots of several
grain crops, forage grasses, and tree crops such as coconut, pepper, and rubber, where it fixes
atmospheric nitrogen. Azospirillum fixes about 20 to 25 kg /ha of nitrogen, therefore, it can
effect a reduction of 25 per cent in the quantity of N fertilizers required. It is considered as a
good bacterium for the tropics as it can withstand high soil temperature. It consumes less
energy and easily colonises on roots and surroundings. It also induces good rooting in target
plants. It is found good for inducing rooting in polybag-raised seedlings of orchards and
plantation crops.
Azospirillum cultures are available in peat or lignite carriers. It is applied as seed
treatment, seedling root dip, and direct soil application. For grain crops, it is applied as
treated seeds. For treating 10 kg seeds, 500g culture is required. Moisten the seeds by
sprinkling water or rice-gruel. Take 500 g culture in a plastic tray and then add moistened


Lecture note by Dr. C. George Thomas , Professor & Head ( Agronomy)& Associate Dean, for the course
Agro 1101 Fundamentals of Agronomy (2+1)

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seeds, mix well, and allow to dry them in shade for 30 minutes. This shall be sown
immediately.
For seedling root dip, slurry of the culture is prepared by adding 50 ml water to 500g
culture and mixing well, and the roots are dipped in the slurry for 15 minutes before
transplanting. For direct soil application, mix the culture with farm yard manure or compost
in the ratio 1:25 and applly directly to the soil.
Azotobacter
Azotobacter is a heterotrophic free-living nitrogen fixing bacterium, which derives
energy from the degradation of plant residues. It fixes about 15 to 20 kg/ha nitrogen under
ideal conditions. A. chroococcum is the most widely distributed species in the tropics in
aerobic soils and plant rhizosphere.
Azotobacter can form beneficial association with most of the crops including cereals,
vegetables, root crops such as cassava, and many tree crops. Requirement of nitrogen
fertilizers can be reduced, if Azotobacter is supplemented, and the quantum depends on the
particular crop. It is often available with carriers. It can be applied in three ways, seed
treatment, seedling root dip and soil application.
Acetobacter
Acetobacter (now Gluconacetobacter) is an endophytic nitrogne-fixing bacteria found
in symbiotic relationships in sugarcane tissues. Gluconacetobacter is the only biofertilizer
capable of surviving in high sugar concentration conditions. Its presence has also been
reported in other plants such as napier grass (Pennisetum purpureum), sweet potato, coffee,
tea, banana, ragi, rice, and pineapple. G. diazotrophicus is the predominant species. This
organism through biological N fixation plays an important role in providing accessible
nitrogen to the plants. G. diazotrophicus carries out nitrogen fixation under aerobic
conditions.
Blue green algae (BGA)
Blue green algae are completely autotrophic. They play a vital role in the nitrogen
economy of rice soils in the tropics. Inoculation of rice fields with BGA has significantly
increased grain yield in many rice cultivars, meeting part of the nitrogen demand. In addition
to nitrogen fixation, BGA secrete vitamins and several growth-promoting substances, which
are beneficial to the crops. BGA fix about 25 to 30 kg of nitrogen in a hectare per season. It
is claimed that about 30 percent of the nitrogen requirement can be met by the use of this
biofertilizer. A soil based mixture of Tolypothric, Aulasira, Nostoc, Anabaena, and
Plectonema is usually used. However, BGA cultures do not perform well in acidic soils with
below 6.0.
AZOLLA CULTURE
Azolla or mosquito fern is a floating aquatic fern (Azolla pinnata) with nitrogen fixing
blue green algae (Anabeana azollae) associated with it. The BGA symbiant is seen in the
central cavity on the dorsal lob of the fronds. The azolla-anabaena association, in addition to
fixing atmospheric nitrogen, gives high yield of biomass too, which can be used as a rich
green manure and good quality feed.
Azolla plant consists of a short, branched, floating stem. The shape is typically
triangular measuring about 1.5 to 3.0 cm in length and 1 to 2 cm in breadth. Roots emanating
from growing branches remained suspended in water. Each leaf is bi-lobed, the upper lobe

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containing green chlorophyll while the lower lobe is colourless. In certain conditions,
anthocyanin pigment also occurs in the plant giving the fern a reddish-brown colour. Often,
this is associated with over-manuring of ponds, pollution, and excess sunlight. Azolla prefers
partially shaded conditions. Azolla is highly productive and is capable of producing 730Mg
of biomass from a pond of one hectare. Azolla multiplies vegetatively at a faster rate.
However, sexual multiplication is also observed.
Azolla is popular as a biofertilizer in South East Asian countries such as China,
Vietnam, and the Philippines. Cultivation of azolla along with paddy and fish is a common
norm in such countries as it increases both rice production and fish production. In the recent
years, there has been a shift in its use away from biofertilizer to a livestock feed, especially as
a substitute for concentrates. In the ponds where it is grown, it can control mosquitoes by
blocking the water-surface. The name “mosquito fern” is because of this mosquito prevention
quality.
MYCORRHIZAE
Some soil fungi called mycorrhizae meaning “fungus roots” is an economically
important, mutually beneficial (symbiotic) association between numerous fungi and roots of
higher plants. This association of fungi and plant roots was first noted on certain forest trees.
It is known to be present in most plant species perhaps excepting the members of family
Brassicaceae and Chenopodiaceae. Mycorrhizal association is of great practical significance,
as it markedly increases the availability of several essential nutrients like P, Zn, Cu, Ca, Mg,
and Fe to plants. Mycorrhizae also increase tolerance of plant seedlings to drought and
extreme soil acidity. Protection from many root pathogens, including fungi and nematodes,
was also reported. In return, plant roots provide the fungi involved with some sugars and
certain organic exudates, which can be used as food.
The most important agriculturally important mycorrhiza is the arbuscular
mycorrhizal fungi or AMF (formerly, vesicular arbuscular mycorrhizae (VAM)). Inside the
cortical cells of the host roots, the fungi form highly branched hyphae called arbuscules.
Mycelia of AMF is both inter and intra cellular and the intercellular mycelia produce
characteristic globose, enlarged, sac-like structures called vesicles, which store reserve food
materials and this later transforms into chlamydospores, the reproductive structures. As these
fungi produce these characteristic structures of arbuscules, they are commonly called
arbuscular mycorrhizal fungi. In Kerala soils, Glomus fasciculatum and G. microspora are
widely seen. The roots of most of the crops such as maize, cotton, wheat, potato, beans,
sugarcane, cassava, upland rice, apples, grapes, citrus, cocoa, coffee, and rubber have AM
associations. AM fungi (AMF) help plants to capture nutrients such as phosphorus, sulphur,
nitrogen, and micronutrients from the soil.
PHOSPHATE SOLUBILIZING MICROORGANISMS
Many soil microorganisms, which include bacteria, fungi and actinomycetes, possess
the ability to bring insoluble phosphates present in the soil into soluble form by secreting
organic acids. This brings about the dissolution of immobilized phosphates. Some of the
hydroxy acids thus produced may chelate with calcium and iron resulting in solubilization
and higher utilisation of soil phosphates. Several members of Bacillus, Pseudomonas,
Micrococcus, Flavobacterium, Penicilium, Fusarium, Sclerotium and Aspergillus were
reported to possess the ability of solubilizing phosphates. They normally grow in a medium
containing insoluble tricalcium phosphates, apatite, rock phosphates, and iron and aluminium
phosphates as sole source of phosphates. In India, Pseudomonas striata, Bacillus polymixa

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and fungi like Aspergillus awamori have been selected for the preparation of carrier-based
inoculants.
Phosphate solubilising microorganisms (PSM) produce both organic (formic, acetic,
citric and oxalic) and inorganic acids (sulphuric and nitric), which convert insoluble
phosphates into monobasic phosphates. As these organisms solubilize more phosphorus than
required for their growth, the surplus will be utilized by the plants. The low grade rock
phosphate available in India can be used directly as a source of low cost P fertilizer along
with phosphate solubilising microorganisms. These are available as carrier based inoculum,
and recommended in neutral and slightly alkaline soils as seed treatment and direct
application. However, in acidic soils, there is no need to use this type of biofertilizer because
of the natural solubilisation of insoluble phosphates happening in such soils.
POTASSIUM AND ZINC MOBILIZING BACTERIA
Potassium exists in soil in different forms but the crop can absorb only what is present
in soil solution. Total K content in soil ranges from 3Mg to 100Mg/ha in the upper 20cm of
soil profile. However, the readily available potassium in soil solution is just 2 per cent out of
the total and the remaining 98 per cent of soil potassium is locked up in soil minerals, and
hence not readily available for plant absorption. This portion of fixed potassium in the clay
minerals could be mobilized by the use of potassium mobilizing bacteria. Potassium
mobilizing bacteria holds much promise for organic farming aspirants. Potassium nutrition is
a great hurdle in organic farming.
A bacterium mobilizing soil potassium (KMB), Frateuria aurentia, has been
identified from the banana rhizosphere soil and advocated for use among the farmers based
on field testing on paddy, groundnut and vegetable crops. Using this KMB, farmers can
mobilize the potassium present in their own field soil and save up to 50 per cent of their
potassium fertilizer requirement. This biofertilizer is suitable for all crops including rice,
sugarcane, fruit crops, vegetables, medicinal and aromatic plants, and many green
houseplants.
Similar to potassium, zinc can also be mobilized by the use of microorganisms. Zinc
mobilising bacteria (ZMP) require inorganic molecules as an electron donor and inorganic
carbon such as carbon dioxide as a source. They obtain nutrients by oxidizing iron and
sulphur with oxygen. Zinc mobilising bacteria of promise is Thiobacillus thioxidans; it
secretes organic acids that help in mobilizing the fixed zinc making it available to the plants.
This also brings down the pH of the soil and helps in reclaiming alkaline oils. It can be used
in crops such as rice, sugarcane, orchards, vegetables, and medicinal and aromatic plants. It is
formulated as carrier based powder. For solubilizing zinc in soil, apply ZMP powder at
25kg/ha in soil with compost.

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