1

What is the effect of Nickel (II) Chloride concentration on the germination rate
(%) of Triticum aestivum seeds?
Introduction

With the ongoing industrialization of the word, abiotic stress factors due to urbanization have
led to the incorporation of pollutants such as pesticides, household chemical waste products,
and heavy metals in natural resources such as soil and water, degrading not only the
environmental quality but entire ecosystems as a whole. Soil is a valuable and non-renewable
resource essential for the growth of plants, thus supporting the majority of ecosystems in the
world. Agriculture is something all human life is dependent upon. Early cities were built around
rivers, as they provided the most fertile soil that would lead to an increase in crop yield. The
presence of various by-products of human activity, such as mining, can have detrimental
impacts on organic life through bioaccumulation. Growing up surrounded by the effects of the
growing climate crisis, the implications of human activities on the environment has been a field
of investigation that I have been interested in for quite a while. Researching the effect of
pollution on crops came to me when my mother told me about how she read an article that said
that the food we eat today is nearly ten times less nutrient-dense than the food that people ate
a decade ago. These figures aren’t very scientifically accurate but reading about how different
factors such as the distance between the crops and the nearest urban region can impact the
nutrient quality and growth of plants inspired me to conduct this experiment.

Investigation

1. Background information:
Bioaccumulation can be defined as the net accumulation of contaminants in organisms, relative
to that in the environment. It is resultant of all uptake and loss processes, including respiration
and dietary uptake, egestion, and metabolism1. The presence of heavy metals, such as nickel,
mercury, and copper in air, soil and water result in bioaccumulation, which ends up affecting
the entire ecosystem, as plants act as primary producers. The ecological impacts that the
bioaccumulation of toxins present include damage to vegetation, soils composition, the
inhibition of revegetation, and the disruption of aquatic ecosystems.

The major sources of heavy metal ion pollution are the overburdening of mines, and the usage
of industrial effluent, fertilizers and pesticides. Metal ions are toxic to the metabolic processes
of plants, mainly through their interaction with the functional groups of molecules in the cell,
especially proteins and polynucleotides. A high concentration of metal ions in soil has been
attributed to reduced shoot and root elongation, oxidative damage, membrane alterations, and
nutrient loss. Germination is the resumption of active growth of the embryonic axis in a seed.

1
Borgå, K. “Ecotoxicology: Bioaccumulation.” Reference Module in Earth Systems and Environmental
Sciences, 2013, doi:10.1016/b978-0-12-409548-9.00765-x. Accessed 15th May 2020.
.
 2

This is indicated by the rupture of the encasing and a visible plant root and is essential to the
propagation of a plant.

Nickel has been shown to be highly toxic to most plant species, as it affects the enzymatic
activity of various essential enzymes, such as amylase, protease, and ribonuclease thus has
been shown to cause retardation of seed germination and growth of many crops2. Furthermore,
the presence of nickel ions has been attributed to reducing plant height, root length, chlorophyll
content3. Nickel stress has also been reported to cause the accumulation of electrolyte ions in
plant stores and impact photosynthetic machinery.4 Triticum aestivum or common wheat is the
most widely grown of all crops and is the cereal with the greatest monetary yield. It has been
widely used in clinal phytotoxicity studies as it is rapidly growing and is relatively easy to
analyse.

2. Aim:
To investigate the effects of varying concentrations of Nickel (II) Chloride (0, 30, 60, 90, 120
mg/L) on the germination rate of Triticum aestivum by observing the number of seeds
germinated over a period of 5 days in solutions of 0-5 days.

3. Hypothesis:
Alternate hypothesis (H1): As nickel ion concentration increases, the germination rate of
Triticum aestivum decreases. This is as a higher nickel ion concentration leads to a greater
interference in the activity of essential ribonuclease and protease enzymes, primarily due to an
accumulation of electrolyte ions in seed stores. Additionally, the bioaccumulation of nickel is
associated with a decline in membrane stability and the mobilisation and digestion of food
reserves in germinating seeds.

Null hypothesis (H0): As nickel ion concentration increases, there is no change in the
germination rate of Triticum aestivum.

4. Experimental variables:
Independent variable: Nickel (Ni+) ion concentration (ppm) (± 0.01ppm)
Solutions of 0, 30, 60, 90, 120 mg/L were used.

Hydrous nickel (II) chloride was used, as this is the most common soluble form of nickel. These
values were chosen in accordance with the nickel ion concentration found in Indian rivers, from

2
Baruah, Nijara, et al. “Influence of Heavy Metals on Seed Germination and Seedling Growth of Wheat, Pea,
and Tomato.” Water, Air, & Soil Pollution, vol. 230, no. 12, 2019, doi:10.1007/s11270-019-4329-0. Accessed
15th May 2020.
3
Chen, Cuiyun, et al. “Functions and Toxicity of Nickel in Plants: Recent Advances and Future Prospects.”
CLEAN - Soil, Air, Water, vol. 37, no. 4-5, 2009, pp. 304–313., doi:10.1002/clen.200800199. Accessed 20th
May 2020.
4
Ghosh, Shyamasree, and Sunilkumar Sethy. “Effect of Heavy Metals on Germination of Seeds.” Journal of
Natural Science, Biology and Medicine, vol. 4, no. 2, 2013, p. 272., doi:10.4103/0976-9668.116964. Accessed
16th May 2020.
 3

a report published by the Central Water Commission5. Concentrations from 0 to 120 mg/L
further allowed for the investigation to explore the effect of varying levels of nickel ion
presence. A concentration of 0 mg/L provides a control group, which can be used as a point of
comparison for the other groups. The control group values were also used in the calculation of
the germination rate.

Dependent variable: The number of seeds successfully germinating over a period of 5 days.

This was used to calculate the germination rate of the Triticum aestivum samples. This was
chosen as the germination rate is used as a measure of the time course of seed germination and
allows us to effectively compare the rates of each batch of seeds. This was calculated using the
following germination rate formula

% 𝐶𝑜𝑛𝑡𝑟𝑜𝑙 𝑔𝑟𝑜𝑢𝑝 𝑔𝑒𝑟𝑚𝑖𝑛𝑎𝑡𝑖𝑜𝑛 − % 𝑇𝑒𝑠𝑡𝑒𝑑 𝑔𝑟𝑜𝑢𝑝 𝑔𝑒𝑟𝑚𝑖𝑛𝑎𝑡𝑖𝑜𝑛

𝐺𝑒𝑟𝑚𝑖𝑛𝑎𝑡𝑖𝑜𝑛 𝑟𝑎𝑡𝑒 =
% 𝐶𝑜𝑛𝑡𝑟𝑜𝑙 𝑔𝑟𝑜𝑢𝑝 𝑔𝑒𝑟𝑚𝑖𝑛𝑎𝑡𝑖𝑜𝑛

Controlled variables:
Variable Reasoning Method of control
100ml of water was measured and provided
The germination of seeds is dependent on the to the initial seed set-up. The same amount
Amount of water
availability of ample amounts of water. (10 ml) of water was additionally added
every day.
Waters from differing sources have different
Distilled water was used from a singular
electrolyte ion concentration and will differ
source to prevent disparity. In addition to
in their hardness, which has been shown to
Source of water this, the same distilled water used for the
impact seed germination.6 Distilled water
control was used to make the NiCl2
lacks additional minerals and electrolytes
solutions.
that could interfere with nickel activity.
The temperature can impact seed
germination by affecting metabolic enzyme Seeds were kept at a constant temperature of
Temperature activities. 25o C was chosen as it has been 25o C by conducting the experiment in a
shown to be the ideal temperature for T. closed room with an air conditioner.
aestivum germination7.
The amount of available light can impact the All seedlings were kept in the same area,
Light available
germination of seeds resulting in no variation between groups.

5
River Data Directorate, Ministry of Water Resources, Government of India, 2014, Status of Trace and Toxic
Metals in Indian Rivers - 2014. Accessed 12th August 2020.
6
Maity, Moumita, et al. “Water Quality Demonstrates Detrimental Effects on Two Different Seeds during
Germination.” Current Agriculture Research Journal, vol. 7, no. 3, 2019, pp. 377–389.,
doi:10.12944/carj.7.3.14. Accessed 12th August 2020.
7
Ceccaroni, Dayana, et al. “Effect of the Time and Temperature of Germination on the Phenolic Compounds of
Triticum Aestivum, L. and Panicum Miliaceum, L.” LWT, vol. 127, 2020, p. 109396.,
doi:10.1016/[Link].2020.109396. Accessed 12th August 2020.
 4

Different sources of T. aestivum seeds will

All seeds were sourced from the same plant
Source of seeds possess differing genetic characteristics
supply store, minimizing this disparity.
which may impact the rate of germination.
The seeds were visually examined, and any
Some seeds may be damaged or have a lower
seeds that appeared to be discoloured,
Quality of seeds mass, which may impact their germination
damaged, or abnormally small were
rate.
removed.

5. Pilot study: Before conducting the final experiment, a pilot study was conducted with
Nickel (II) chloride solutions of 80 mg/L, 120 mg/L, 160mg/L, and 210 mg/L in order
to ascertain the value at which germination activity was entirely inhibited. This was
found to be the solution at 160 mg/L; thus 120 mg/L was chosen as the maximum
solution concentration.

A self-taken image showing the setup for the pilot study

Methodology

1. Materials:
- Glassware:
Uncertainties
Least count Number used in
Name of apparatus Volume (ml) (if used for
(ml) one trial
measurement) (ml)
Petri dishes - - - 10
250ml graduated
250 2 ±0.400 5
cylinder

1l beaker 1000 - - 5

50ml graduated
50 5 ±0.100 5
cylinder
 5

- Apparatus:
× Graduated pipette (1)
× 10.0 cm ruler (±0.050 mm)
× Filter paper
× Electric mass balance (±0.001g)
× Stirring rod (1)

- Chemical and biological materials:

× Triticum aestivum seeds (360 seeds)
× Nickel (II) Chloride (840 mg)
× Distilled water (15 litres)

2. Procedure:
Preparation of seeds and beakers
a) Using the ruler, measure the length of the Triticum aestivum seeds and pick 10 seeds
around 0.7cm in width.
b) Leave all seeds in a petri dish filled with distilled water overnight.
c) Fill 6 1l beakers with distilled water up to the 1l marked line
d) Label the beakers for each of the concentrations required (0, 30, 60, 90, 120 mg/L)

Preparation of solutions
The following steps e) to g) were repeated for each NiCl2 solution.
e) Weigh 30 mg of anhydrous NiCl2 using the mass balance.
f) Transfer the weighed powder to its cognate 1l beaker.
g) Using a stirring rod, stir the solution until no visible sediment can be seen.

Preparation of Petri dishes

h) Label 5 Petri dishes from A to E, corresponding to all 5 concentrations.
i) Measure 25ml of each NiCl2 solution. Pour these into labelled 50ml beakers
numbered from 1-5.
j) In a petri dish, cut filter paper to size and layer 4 sheets of filter paper to provide a
base for seed germination.
k) Pour the corresponding solution into each petri dish. Using a stirring rod, press
down the sheets so that the entire surface is saturated with the NiCl2 solution.
l) Place 10 Triticum aestivum seeds on the surface of each petri dish.
m) Place the 5 dishes in a sunny area close to each other.

Data collection
n) Wait for 5 days. Each day add 10ml of the corresponding solution to each petri dish.
o) Count the number of seeds with a visible shoot longer than 10mm in length.
p) Repeat the experiment thrice with each concentration to improve the reliability of
data.
 6

The seeds were observed for a period of a week, to allow for sufficient time to gauge the
effectiveness of each solution of varying Ni+ ion concentration on the growth rate of the
seedlings. Since the Petri dishes were placed next to each other, and trials were conducted
simultaneously, they are assumed to be receiving equal amounts of light. One method of data
collection for this experiment was to count the number of seeds that germinated successfully.
This was done by observing whether the seed coat of each seed was broken and whether the
seedling had emerged. The statistical significance between Ni+ ion concentration and seed
germination were found using the χ2 test.

3. Risk assessment:
Safety considerations: Nickel (II) chloride is toxic if ingested and can be a
respiratory/dermatological irritant, thus the experiment was conducted using latex gloves and
a lab coat to ensure safety.

Environmental considerations: Nickel (II) chloride has long term adverse impacts on aquatic
ecosystems; thus, it was not emptied down a sink and was disposed of following hazardous
waste protocols.

Ethical considerations: There were no ethical considerations to take into account.

Raw Data

1) Qualitative Data:
• The solutions with a higher concentration of nickel (II) chloride had a slight blue
tint.
• The seed coats of seeds in the 0 and 30 mg/L solutions showing signs of germination
as soon as the second day.
• The seeds in the 0 mg/L solution were already showing steady shoot growth at the
end of the week.
• The shoots of the seeds in the 90 and 120mg/L groups showed minor irregularities
in form compared to those in the 0 mg/L groups.
• The seeds in the higher concentration groups appeared to take on the blue tint of the
nickel (II) chloride solutions by day 4.
 7

2) Quantitative Data:

Data Table 1: A raw data table containing the number of seeds germinated in each trial
Concentration of Number of seeds germinated (/20)
Nickel ions
Trial 1 Trial 2 Trial 3 Trial 4 Trial 5 Trial 6
(mg/L)
0 19 20 20 21 19 20
30 16 19 17 16 18 16

60 14 16 14 15 13 14

90 13 11 10 12 11 12

120 9 12 10 10 8 9

Data Processing

Table 1 shows an inverse correlation between the concentration of the NiCl2 solution and the
number of germinated seeds. In order to better understand this relationship and coherently
display the collected data, the mean of the values from each trial will be determined.
Furthermore, the standard deviation of these values will be determined in order to ascertain
the dispersion within the dataset.

The average number of seeds germinated was calculated using the following formula:

𝑆𝑢𝑚 𝑜𝑓𝑠𝑒𝑒𝑑𝑠 𝑔𝑒𝑟𝑚𝑖𝑛𝑎𝑡𝑒𝑑 𝑖𝑛 𝑒𝑎𝑐ℎ 𝑡𝑟𝑖𝑎𝑙

= 𝐴𝑣𝑒𝑟𝑎𝑔𝑒 𝑛𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑠𝑒𝑒𝑑𝑠 𝑔𝑒𝑟𝑚𝑖𝑛𝑎𝑡𝑒𝑑
120

The standard deviation of values was calculated using the following formula:
∑!"# ' "
$%! ($$ %&)
𝜎=# )*+

Data Table 2: A processed data table containing the averages and standard deviation of the
number of seeds germinated at each nickel ion concentration.
Concentration of Nickel Average number of Standard deviation of
ions (mg/L) seeds germinated (0/20) values
0 19.83 0.687
30 17.33 1.105
60 14.34 0.942

90 10.23 1.061

120 9.01 0.816

 8

Graph 1: A graph showing the average number of seeds germinated at each nickel ion
concentration showing the standard deviation through error bars
25

20
Mean number of seeds germinated

y = -0.0959x + 19.898
R² = 0.9812
15

10

0
-20 0 20 40 60 80 100 120 140
Concentration of Nickel (II) Chloride solution added (mg/L)

This graph, in addition to the data depicted in Table 2, supports the alternative hypothesis, as
they showcase a negative correlation with the concentration of Nickel ions. To better
represent the effects of Nickel ion concentration on seed germination, the germination rate
will be calculated.
𝑁𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑠𝑒𝑒𝑑𝑠 𝑔𝑒𝑟𝑚𝑖𝑛𝑎𝑡𝑒𝑑
𝐺𝑒𝑟𝑚𝑖𝑛𝑎𝑡𝑖𝑜𝑛 𝑟𝑎𝑡𝑒 =
𝑇𝑜𝑡𝑎𝑙 𝑠𝑒𝑒𝑑𝑠

Data Table 3: A processed data table containing the calculated germination rate of Triticum
aestivum seeds at each nickel ion concentration
Concentration of Nickel ions Average number of seeds
Germination rate (%)
(mg/L) germinated
0 19.83 99.15
30 17.33 86.65
60 14.34 71.70
90 10.23 51.15
120 9.00 45.00

The calculated germination rates further support the alternative hypothesis, in order to better
depict the relationship between nickel ion concentration and germination rate, a scatter graph
is plotted, with error bars at a standard value of 5%.
 9

Graph 2: A graph of the germination rate of Triticum aestivum seeds at each nickel ion
concentration
120

100
y = -0.4793x + 99.49
R² = 0.9812
Germination rate %

80

60

40

20

0
0 20 40 60 80 100 120 140 160
Concentration of Nickel (II) Chloride solution added (mg/L)

To ascertain the variability of the data collected, the percentage error derived from the
equipment utilized will be calculated.

Margin of error in the mass balance: ±0.001g

Margin of error in the 250 ml graduated cylinder: ±0.5ml
Margin of error in the 50 ml graduated cylinder: ±0.4ml

25ml of each NiCl2 was measured using the 50ml graduated cylinder, thus the percentage
uncertainty for the same would be the following:

0.4
× 100 = 1.60 %
25

The mass balance used to weigh the NiCl2 required to make each solution had a margin of
error of ±0.001g. The smaller the measured value, the higher the uncertainty. Substituting the
minimum measured mass (30mg) into the same we get the following:

0.001
× 100 = 3.33%
0.03

The 250ml graduated cylinder was used to adjust the NiCl2 solutions to the final volume of 1
litre. The 250ml graduated cylinder has a margin of error of ±0.5ml.

0.5
× 100 = 0.2 %
250
Adding together all the uncertainties, we get the maximum percentage uncertainty (%) of
5.12%
 10

Data Analysis

A maximum percentage uncertainty of 5.12%, as well as the standard deviation values, suggest
an acceptable level of variance and uncertainty within the dataset. The R2 value of both Graph
1 and 2 is 0.981. The R2 value is a measure of how well a linear regression model fits a dataset.
These values range from 0 to 1. A value of 0.98 indicates a strong relationship between the
model and the dependant variable. Graph 2, as well as the difference in percentage germination,
falls in line with the alternative hypothesis, showing a negative correlation between nickel (II)
chloride concentration and percentage germination, however, to test the statistical significance
of this relationship, the ANOVA test was conducted.

The following working hypotheses were used in the conduction of the ANOVA test:
H0: There is no difference in the means of the five concentration groups.
H1: There is a significant difference in the means of the five concentration groups.

Table 10 (From Microsoft Excel): A table displaying the input and results of the ANOVA test.

The p-value was found to be below the accepted significance limit of 0.05, which shows that
the differences in the averages of the five sets of values are similar, further, a significance level
below <0.001 suggests that the relationship between the variables are highly statistically
significant. Thus, the null hypothesis can be rejected and the alternative hypothesis, stating that
there is a significant difference between the means of NiCl2 concentration and the germination
of Triticum aestivum seeds.
 11

Evaluation

Table 11: A table displaying possible sources of errors and improvements

Error Type of Significance Method of Improvement
error
Process used Germination could have also been
The criteria of a visible shoot longer than
to determine Systematic monitored through enzyme activity as
10mm in length were used as a standard in
germination error a more accurate means of determining
order to ensure accurate data.
of seeds germination activity.
Cotton was used as a substrate for the
Substrate Systematic germinating seeds. Soil and treated potting Usage of a treated potting mixture for
utilised error mixtures contain essential nutrients that germinating seeds.
assist in seed germination.
The NiCl2 solutions were created using
distilled water, which lacks essential
Usage of Bottled drinking water could have
minerals which play a role in regulating
distilled Systematic been used in order to have a source of
hydrolytic enzyme activity8. However, the
water in the error essential minerals at a controlled
usage of tap water may have impacted the
solution concentration.
activity of nickel ions and would have been
difficult to standardise.
Measurement The approximate diameter of the seeds was
of the measured with a ruler prior to usage in the The mass of the seeds could have been
Systematic
Triticum study, which is not a very accurate method measured in order to reduce any
error
aestivum of determining the viability of the seeds disparity between the seeds.
seed size utilised.
Germination is reliant on light availability The experiment could have been
and enzyme activity, which is sensitive to conducted in a room with more
Control of temperature. All samples were placed in an ventilation and a better temperature
Random
external area receiving the same amount of natural control system. Furthermore, lamps
error
variables light and ventilation at a controlled could have been used as a light source
temperature, the experimental conditions in order to reduce any disparity due to
could have been further controlled. light availability.
This paper only utilised primary data as a Secondary data taken from databases
source of evaluation. Although the data showcasing regional germination rates
acquired is proved to be of statistical value, or crop yields could have been
Sources of
Limitation the usage of secondary data could have compared with soil data gathered from
data
assisted in understanding the relationship agricultural surveys in order to
between nickel ion concentration and seed ascertain whether the same correlation
germination. is present in an in-vivo setting.
As discussed in the Background Research
section, nickel stress has been attributed to
the reduction of plant height, root length,
Studying the growth of the seeds over
Frame of and chlorophyll content. Studying these
Limitation the period of 2-3 weeks instead of a
observation variables would also allow us to gain a
singular week.
deeper understanding of the impact of
nickel stress on long-term plant welfare,
instead of just seed germination.

8
Ali, Awatif S., and Alaaeldin A. Elozeiri. “Metabolic Processes During Seed Germination.” Advances in Seed
Biology, 2017, doi:10.5772/intechopen.70653. Accessed 16th August 2020.
 12

Conclusion

The experiment shows that nickel stress can dramatically impact Triticum aestivum seed
germination rates. The germination rate observed for the control was 99.15%, whereas the rate
observed for the highest nickel concentration used (120mg/L) was 45.00%. Although some
improvements could have been made to improve the quality of the data obtained, an assessment
of the measures of uncertainty and variance, as well as the ANOVA test show that the data is
of statistical significance, allowing us to reject the null hypothesis and accept the alternative
hypothesis. As illustrated in graph 2, the relationship follows an inverse correlation, with an
increase in NiCl2 concentration resulting in a decreased germination rate, in line with the
alternative hypothesis presented. The standard deviation values are shown in Table 2 suggest
a low level of random error in the experiment, which evidences the accuracy of the data
collected. The maximum percentage uncertainty was found to be 5.12%, which is just over the
value of 5% typically accepted within scientific research. This can be attributed to the low level
of precision of the mass balance used. The high R2 value of graph 2 indicates a strong inverse
correlation between the concentration of NiCl2 in solution and the germination of Triticum
aestivum seeds. This can be attributed to the toxicological effects that that nickel ions have on
the enzyme activity and photosynthetic machinery required for seed germination. Nickel stress
has also been found to affect the digestion and mobilisation of energy reserves in germinating
seeds, and the membrane stability of growing plants. Furthermore, the bioaccumulation of
nickel ions has been known to impact the activity of ribonuclease and protease enzymes,
retarding seed germination and causing a decline in growth, membrane stability, and lipid
peroxidation4. This can be seen in the data collected since the rate of germination of seeds in
the 0 mg/L groups was 54.15% higher than that of seeds in the 120 mg/L group. This
additionally explains why the shoots emerging from the seeds in the 60-120 mg/L groups
showed slight irregularities in form.

Further Scope: Nickel is an essential nutrient for plants, as it plays a role in regulating tissue
urea levels and antioxidant metabolism9, however understanding its impact as a phytotoxin as
a side effect of mine exhaustion and industrial pollution is important to consider in a time of
great ecological strain, where many countries are facing devastation due to agricultural issues
and a lack of nutrients. Broadening the scope of the experiment, studying the impact of various
heavy metal salts on seed germination, would be a means of delving deeper into the interactions
of these ions with seeds, allowing us to better understand how researchers can manipulate
pathways to prevent impact on agricultural produce as well as human health in the long-term
in order to combat the strain put on agricultural sectors by the reduction in soil quality.

9
Fabiano, Caio C., et al. “Essentiality of Nickel in Plants: a Role in Plant Stresses.” Frontiers in Plant Science,
vol. 6, 2015, doi:10.3389/fpls.2015.00754. Accessed 16th August 2020.
 13

Works Cited
Ali, Awatif S., and Alaaeldin A. Elozeiri. “Metabolic Processes During Seed Germination.” Advances
in Seed Biology, 2017, doi:10.5772/intechopen.70653. Accessed 16th August 2020.

Baruah, Nijara, et al. “Influence of Heavy Metals on Seed Germination and Seedling Growth of Wheat,
Pea, and Tomato.” Water, Air, & Soil Pollution, vol. 230, no. 12, 2019, doi:10.1007/s11270-019-
4329-0. Accessed 15th May 2020.

Borgå, K. “Ecotoxicology: Bioaccumulation.” Reference Module in Earth Systems and Environmental

Sciences, 2013, doi:10.1016/b978-0-12-409548-9.00765-x. Accessed 15th May 2020.

Ceccaroni, Dayana, et al. “Effect of the Time and Temperature of Germination on the Phenolic
Compounds of Triticum Aestivum, L. and Panicum Miliaceum, L.” LWT, vol. 127, 2020, p.
109396., doi:10.1016/[Link].2020.109396. Accessed 12th August 2020

Chen, Cuiyun, et al. “Functions and Toxicity of Nickel in Plants: Recent Advances and Future
Prospects.” CLEAN - Soil, Air, Water, vol. 37, no. 4-5, 2009, pp. 304–313.,
doi:10.1002/clen.200800199. Accessed 20th May 2020.

Fabiano, Caio C., et al. “Essentiality of Nickel in Plants: a Role in Plant Stresses.” Frontiers in Plant
Science, vol. 6, 2015, doi:10.3389/fpls.2015.00754. Accessed 16th August 2020.

Ghosh, Shyamasree, and Sunilkumar Sethy. “Effect of Heavy Metals on Germination of Seeds.” Journal
of Natural Science, Biology and Medicine, vol. 4, no. 2, 2013, p. 272., doi:10.4103/0976-
9668.116964. Accessed 16th May 2020.

Maity, Moumita, et al. “Water Quality Demonstrates Detrimental Effects on Two Different Seeds
during Germination.” Current Agriculture Research Journal, vol. 7, no. 3, 2019, pp. 377–389.,
doi:10.12944/carj.7.3.14. Accessed 12th August 2020.

Parlak, Kadiriye Uruç. “Effect of Nickel on Growth and Biochemical Characteristics of Wheat
(Triticum Aestivum L.) Seedlings.” NJAS - Wageningen Journal of Life Sciences, vol. 76, 2016,
pp. 1–5., DOI: 10.1016/[Link].2012.07.001. Accessed 24th July 2020.

Pokorska-Niewiada, Kamila, et al. “Acute Lethal Toxicity of Heavy Metals to the Seeds of Plants of
High Importance to Humans.” Bulletin of Environmental Contamination and Toxicology, vol.
101, no. 2, 2018, pp. 222–228., doi:10.1007/s00128-018-2382-9. Accessed 24th July 2020.

River Data Directorate, Ministry of Water Resources, Government of India, 2014, Status of Trace and
Toxic Metals in Indian Rivers - 2014. Accessed 12th August 2020.