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The Effect of Fertilizer on Lead Resistance in Brassica rapa

Brianne Cook, Samantha Eckrich, Micaela Fosdick, and Tyler Gabrelcik
Loras College
1450 Alta Vista St.
Dubuque, IA 52001
Chemical pollution can have detrimental and sometimes fatal effects on grasses, trees and other flora
surrounding sites of accidental and intentional contamination. Through this experiment, we explored the
use of fertilizer to counteract the harmful influence of a pollutant, lead nitrate. The growth of 180
Brassica rapa plants was observed and measured within nine carefully-controlled experimental groups
exposed to varying nutrient and pollutant concentrations. After four weeks, we removed the plants from
the soil and measured the biomass of the individual plants. Our initial hypothesis that plants exposed to
the lowest concentration of lead nitrate and the highest concentration of fertilizer was not supported by
the results of our experiment. Our results suggest that higher concentrations of the fertilizer do not
counteract the effects of the lead pollutant, and that too much could be harmful. We also noticed that the
combination of increasing fertilizer and lead concentration tended to produce plants of lower biomass.
These results, however, may be considered inconclusive because of limited sample size, and thus we have
proposed ideas for improving the experimental design.

Industrial processes often lead to harmful levels of heavy metal contamination in soil.
One such contaminant is lead. Lead is toxic to many organisms, including plants and the animals
that feed on those plants (EPA, 2012). A study done on the plant Arabidopsis thaliana,
comparing cadmium and lead concentrations on biomass, showed lead to be an effective toxin
and significantly reduced plant growth in the experimental group (Yu et al., 2012). Another study
revealed increased lead concentrations contributed to a decrease in total biomass of plants
(Kosobrukhov et al., 2004). Wisconsin fast plant (Brassica rapa) is within the same family as
Arabidopsis, Brassicaceae. It has a similar lifestyle and is a fast-growing plant, making it an
ideal subject for a four-week study examining the effects of a contaminant (such as lead nitrate)
on plant growth. In addition, many studies have shown nutrient supplementation to be a factor in
resistance to heavy metals such as lead in Arabidopsis (Gadepalle et al., 2007). Many
household fertilizers containing the compound urea, a source of nitrogen, as well as other
essential nutrients involved in plant growth (Scotts-Sierra, 2003).
We tested the effects of fertilizer on lead toxicity resistance, measured by plant biomass.
Our hypothesis was that if fertilizer was provided to plants growing in polluted soil, then the
effects of the pollutant will decrease compared to plants without the fertilizer supplement. This
experiment answered the question: What is the influence of fertilizer on Brassica rapa plants
exposed to varying levels of lead nitrate? Our prediction, based on the information gathered from
the sources described above, was that the group of plants exposed to the highest level of fertilizer
and lowest level of lead contaminant would have the greatest biomass at the end of the four-week
period. We also predicted that the plants exposed to the highest level of lead nitrate and no
fertilizer would have the lowest biomass at the end of the four-week period.
Our materials included 180 Brassica rapa seeds, Peters Professional Water Soluble
Fertilizer (150 ppm and 300 ppm), lead nitrate solutions (200 ppm and 500 ppm), distilled and
tap water, potting soil, and 180 individual plastic plant containers.
We added the soil to the individual plastic containers then added one seed to each. We
divided the plants into nine groups with varying fertilizer and lead concentrations (Table 1). Each
group was placed in a different plastic trey for watering purposes.

Table 1. Lead and Fertilizer Treatment Groups (20 plants per treatment)
No Lead

No Fertilizer

Low Lead Content

High Lead Content

(200 ppm)

(500 ppm)


200 ppm lead,

500 ppm lead,

(no lead, no

no fertilizer

no fertilizer

200 ppm lead,

500 ppm lead,

Low Conc. of Fertilizer
(150 ppm)
High Conc. of Fertilizer
(300 ppm)

No lead,
150 ppm fertilizer

150 ppm fertilizer

150 ppm fertilizer

No lead,

200 ppm lead,

500 ppm lead,

300 ppm fertilizer

300 ppm fertilizer

300 ppm fertilizer

We kept the plants in the greenhouse to maintain consistent growing conditions, including
temperature and level of humidity. Every other day, we refilled the water supply in each tray with
pure water for the unpolluted groups and with the corresponding lead solutions for the polluted
groups. Once a week, we added 1 mL of fertilizer to each of the plants requiring either the low or
high concentrations of fertilizer. We observed and recorded the germination of the plants, and
after three weeks we removed the plants and measured their individual masses fresh out of the
Of the 180 Brassica rapa seeds planted, 67 germinated and 43 of these plants continued
to grow until the end of the fourth and final week. Some of the observational differences among
the treatment groups included variance in leaf growth, root systems, and flowering. The control
group had the most complex root systems and three flowering plants; the low lead treatment
group had one flowering plant; the high nutrient treatment group had plants with secondary
leaves; high lead had one flowering plant; low nutrient had two flowering plants. No plants
survived in the high fertilizer/high lead treatment group.
Fertilizer does not have a significant effect on counteracting the lead toxins influence on
the biomass of plants. A two-way analysis of variance tested the biomass of plants treated with
varying levels of fertilizer (F2, 35 = 1.796, p = 0.181) and lead (F2, 35 = 3.024, p = 0.061). Neither

variable had a significant effect on plant biomass (Table 2). However, Figure 1 displays that
although the statistical results were not significant, a general pattern exists between the
concentrations of lead and fertilizer, and biomass: higher concentrations of lead and fertilizer
resulted in lower biomass. Further analysis comparing germination between treatment groups
using a chi-square test reveals a significant difference in germination between treatment groups
( df = 4, 2 = 10.84, 0.01 < p <0.05). The high nutrient and low nutrient treatment groups had
more plants germinate than expected, and fewer plants than expected germinated in the high
lead/high nutrient treatment group.

Table 2. ANOVA Table Comparing the Effects of Lead and Fertilizer Treatments on
Brassica rapa Plants
Tests of Between-Subjects Effects
Dependent Variable:Biomass

Type III Sum of



Mean Square












.030 and Fertilizer
1. The Effect of Lead
Lead * Nutrient










Brassica rapa

a. R Squared = .519 (Adjusted R Squared = .409)



Figure 2. The Effect of Lead and Fertilizer Treatments on Germination ofBrassica rapa

The results of the statistical tests suggest that exposure to high concentrations of fertilizer
does not significantly counteract the effects of lead pollutant. These results do not support our
initial hypothesis that if fertilizer was applied to plants growing in lead-polluted soil, then the
effects of the pollutant would be reduced compared to the plants that did not receive fertilizer.
Increased concentrations of both lead and fertilizer did, however, result in a general trend of
decreased biomass, as observed in Figure 1. The decrease in biomass of plants due to increased
lead concentrations concurs with previous research (Kosobrukhov et al., 2004). However, these
results do not support the findings of studies which suggest that nutrient supplements contribute
to resistance of heavy metals (Gadepalle et al., 2007). This resistance was not observed in our
experiment; fertilizer did not counteract the effects of the lead pollutant, leading to the gradual
decay of some plants.

Fertilizer, however, had an effect on the germination of the plants. The chi-square
analysis suggests germination was significantly different between treatment groups. Treatment
groups exposed to only high and low nutrient concentrations had significantly more plants
germinate than expected (Fig. 2). Fewer plants germinated than expected in the presence of both
lead and nutrient (Fig. 2).These results contradict the findings of previous studies and suggest
that while fertilizer may aid in plant growth independently of a toxin, it does not counteract the
effect of the lead pollutant. Along with this consideration, not all of the plants that germinated
survived the full four weeks. Within all groups except the low lead/high nutrient group, more
plants germinated than survived the four weeks of the experiment. This suggests that fertilizer
may only benefit the plant to a limited degree. This is also indicated in the high lead/ high
nutrient treatment group, in which no plants survived. Furthermore, the higher success rate of the
low nutrient treatment group in comparison to the control also supports the limited benefit of
fertilizer (Fig. 2). The results of this experiment could be considered inconclusive because of the
limited sample size. The chi-square analysis suggests that different treatments did have an effect
on germination. However, possibly because of small sample size, this was not reflected in the
ANOVA test.
Several sources of error may have contributed to the high fatality rate in our experimental
plants. For example, the concentrations of fertilizer, particularly the high-concentrate fertilizer,
and the early, frequent application of the fertilizer may have had a lethal effect. Fertilizers
containing nitrogen may contribute to issues related to nitrate pollution in high amounts (Byrnes,
2009). Furthermore, in the initial week of the experiment, we added lead to the plant trays every
other day and watered the plants inconsistently because of scheduling complications. During the
following three weeks, we arranged more consistent conditions and maintained a continuous
supply of lead nitrate in the trays. This continuous supply of liquid to the plants, however, may
have been detrimental to plant growth by over-watering, and perhaps even contributed to the
number of plant fatalities, and thus ultimately distorting the results of our experiment. Error may
have also been possible in finding the biomass of each of the plants, as we did not allow the
plants to dry before weighing them.
If this experiment were to be repeated, adaptations would have to be made. There would
need to be a decrease of fertilizer concentrations to a more appropriate amount for the plants and
an increase in the amount of time between fertilizer treatments. We would also modify the

technique with which we watered our plants. The instruction booklet supplied with the Brassica
rapa seeds suggests using a water mat and wicks to provide water to the plants rather than
having direct contact between the soil and water source (Carolina Biological Supply Company,
2001). This would be a more effective method of watering and would prevent the possible error
contributed by over-watering.
Further experimentation exploring this topic could include testing various other
pollutants, fertilizer supplements, and species of plants. Many studies have successfully found
organic substrates to assist in heavy metal resistance like manures, sawdust, wood ash, composts,
sewage sludge, and woodchips (Gadepalle et al., 2007). These are other potential experiments
that could supplement our investigation.
We would first like to thank Dr. Daniel Wenny, who helped us design and guide our
experimental procedure as well as obtain the materials necessary for our experiment. We would
also like to extend our gratitude to Mary Jo and Tony for their assistance in producing the lead
nitrate solutions for our experiment. In addition, we would like to recognize Dr. Aditi Sinha, who
offered us advice on making the fertilizer solutions.

Literature Cited
Byrnes, B. 1990. Environmental effect of N fertilizer useAn overview. Fertilizer Research 26:209-215.
Carolina Biological Supply Company. 2001. Wisconsin Fast Plants Growing Instructions.
Burlington, NC: Author.
EPA (United States Environmental Protection Agency). 2012. An introduction to air quality: lead
(Pb). United States Environmental Protection Agency. Accessed September 21, 2013.
Gadepalle,V., S. Ouki, R. Herwijnen., T. Hutchings. 2007. Immobilization of heavy metals in
soil using natural and waste materials for vegetation establishment on contaminated sites. Soil &
Sediment Contamination 16:233-251.
Kosobrukhov, A., I. Knyazeva, V. Mudrik. 2004. Plantago major plants responses to
increase content of lead in soil: growth and photosynthesis. Plant Growth Regulation
Scotts-Sierra Horticultural Products Company. 2002. Peters Professional Water Soluble
Fertilizer 20-20-20 General Purpose. Marysville, OH: Author.
Spittler, M. & W. Feder. 1979. A study of soil contamination and plant lead
uptake in Boston urban gardens. Communications in Soil Science and Plant Analysis 10:9, 1195121.
Yu, B., X. Bian., S. Cao., X. Chen., J. Qian., R. Wang. 2012. Arabidopsis desaturase 2 gene is
involved in the regulation of cadmium and lead resistance. Plant Soil 358:289-300.