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
Abstract
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 worked to
explore 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.

Introduction
Industrial processes often lead to harmful levels of heavy metal contamination in soil.
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, 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). Peters Professional Water Soluble Fertilizer is a common
household fertilizer containing the compound urea, a source of nitrogen, as well as other
essential nutrients involved in plant growth (Scotts-Sierra, 2003).
This experiment tested the effects of Peters Professional fertilizer on lead toxicity
resistance, measured by biomass. Our hypothesis was that if fertilizer was provided to plants
growing in polluted soil, then the effects of the pollutant will differ from 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 Peters Professional fertilizer would have the least biomass at
the end of the four-week period.
Methods
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 plant containers.
We added the soil to the individual plant containers then added the seeds. Using nine
plant trays and the individualized plant containers, we divided them into three groups containing
no lead, low lead content (200 ppm), or high lead content (500 ppm), and within each of these

groups no fertilizer, a low concentration of fertilizer (150 ppm) or a high concentration of

fertilizer (300 ppm) was applied to the plants.
Table 1. Plant Sample Groups
No Lead

No Fertilizer

Low Lead Content

High Lead Content

(200 ppm)

(500 ppm)

Control

200 ppm lead,

500 ppm lead,

(no lead, no

no fertilizer

no fertilizer

200 ppm lead,

500 ppm lead,

fertilizer)
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 plants water supply with
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.

Results

Table 2.
Tests of Between-Subjects Effects

Dependent Variable:Biomass
Source
Type III Sum of
Corrected Model

Squares
.075a

df
7

Mean Square
.011

F
1.261

Sig.
.298

Lead

.051

.026

3.024

.061

Nutrient

.030

.015

1.796

.181

Lead * Nutrient

.001

.000

.031

.992

Error

.297

35

.008

Total

.617

43

Corrected Total

.372

42

R Squared = .201 (Adjusted R Squared = .042)

Figure 1.
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)
Figure 2.

and lead (F2,35 =

0.61) . 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: a decrease in biomass corresponds to the increase of concentrations of both lead and
fertilizer. 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). oppose
Discussion
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 differ from the plants that did not receive fertilizer. Increased
concentrations of both lead and fertilizer did, however, result in a general trend of decreased
biomass (Figure 1). The chi-square analysis suggests germination was significantly different
between treatment groups. Along with this consideration, not all of the plants that germinated
survived the full four weeks. These observations indicate that fertilizer only played a role in lead
resistance prior to germination.
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 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 (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, 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 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.

Acknowledgements
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. We also express our sincerest gratitude for
those seedlings who chose to overcome the barriers of chemical pollution and fully participate in
our experiment; thank you to those twenty-some seeds.

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.
<http://www.epa.gov/iaq/lead.html>.
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
42:145-151.
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.