Density and Competition Effects on Brassica oleracea and Raphanus sativus

Schyler Kidd
November 11th, 2012

Abstract

A plant’s growth ability is dependent on its ability to acquire the resources it needs to
survive. Competition such as interspecific and intraspecific, limiting resources, and
population density affect the fitness level of a plant. This experiment was conducted in
order to test the capability of collards and radishes to grow in manipulated densities under
interspecific and intraspecific competition. I hypothesized that both collard and radish
plants will grow more efficiently in single species pots under low density conditions. I
also hypothesized that in the mixed species pots the radishes will be more fit to survive
and grow better than the collard plants in both the high and low density pots. Both high
and low density, and single and mixed species pots were planted and results were
observed. There was a significant difference found in the in the low and high-density
single species pots; however, there was not a significant difference found in the low and
high density mixed species pots. Experiments are needed to further analyze these results.

Introduction

The effects on plants such as overcrowding and population density have sparked the
interest of scientists over the past several decades. New plant and animal species are
being discovered, and it is vital that we continue to learn about the effects of growing
populations and how they contribute to their advancement. It has been shown by
previous experiments that the limiting amount of resources and nutrients affect the
growth rate and number of populations (Hansen et al 1999). As natural selection has
showed us, some plants and animals are more fit when competing for food, water, light,
space, and nutrients in order to survive (Ricklefs et al 2010). A determining factor in a
plant’s survival rate is its ability to battle for resources. Interspecific competition is when
two species that are not alike are competing for the same resources while intraspecific
competition is when species that are alike are competing for the same resources (Tilman
et al 1997). A plant’s capacity to compete against other species is directly proportional to
its fitness level. If a plant is more fit, it will be able to obtain more nutrients, and thus
grow at a suitable rate (Brown et al 2008).

The main purpose of this experiment was to test the effects of interspecific competition
and intraspecific competition between two species of plants. These species are collard
plants, Brassica Oleracea, and radish plants, Raphanus sativus. Intraspecific competition
was tested among separate species in both high and low-density pots in order to
determine conditions that were favorable for each species. For intraspecific competition, I
hypothesized that both collards and radishes would produce more biomass in low-density
pots versus high-density pots because there will be fewer plants to compete against. The
limiting resources that are manipulating my hypothesis are water and nutrients that each
plant obtains from the soil.

Interspecific competition was tested among species by making mixed species pots that
were both low and high density. I hypothesized that radishes will outgrow the collard
plants due to the size of their root area. Below ground biomass will be overtaken by the
radishes, which will in turn prevent the collards from obtaining the nutrients they need
and therefore allowing the radishes to grow more sufficiently (Hansen et al 1999).

The limiting resources that affect my hypothesis are the water and nutrients the plants
obtain from the soil. In pots with intraspecific competition, the same species will be
competing for the below ground nutrients. This hypothesis was chosen because in the
low-density pots plants will have more space the attain nutrients that are necessary for
their growth and development. In the interspecific competition pots, species that are
different will be competing for the same nutrients and therefore one species will obtain
more nutrients than the other which will be disposed of their level of fitness (Tilman et al
1997). Radishes have a larger root area and will be capable of obtaining nutrients at a
faster rate than collards (Ricklefs et al 2010).

This competition experiment is exploited because it is important to analyze the different

conditions, which determine the success of different types of species of plants. Their
ability to contend against others for resources gives us an idea of their capabilities in the
wild amongst all other types of species. We hope to accumulate more knowledge about
the different effects of plant competition on growth. These results can have high yields
for not only farming techniques, but also for knowledge about plant competition amongst
various species.

Materials and Methods

Field Procedure

The majority of this experiment took place at the Greenhouse 2, located on the corner of
South Main Street at the University of South Carolina. We planted pots using the seeds of
Raphanus sativus (radish plants) and Brassica oleracea (collard plants) to test the effects
of competition on two different types of species. This competition was explicitly
intraspecific and interspecific. The pots were planted using three different levels of
species; collard only, radish only, and radish-collard mix. In addition, two density levels
were used; high density with 64 seeds and low density with 8 seeds.

High and low density pots were planted using radishes, collards, and a radish-collard mix.
The low-density radish-collard mix pots contained four seeds of radishes and four seeds
of collards. The high-density radish-collard pots contained 32 seeds of each species.
While our group replicated this 3x2 design four times to total 24 pots, we incorporated
the whole class data. Therefore, there were 16 replicates for each treatment. For each pot,
we filled soil up until about one inch from the top. We placed the seeds in the pot, and
piled on around 2 or 3 cm of soil on top. In 3 species levels, seeds were spaced as evenly
as possible. In the mixed species pot, the two species were alternated so that each one had
the same access to space and nutrients at the other. For each pot, we wrote down our
section number, group name, and the contents of the pot. Our group worked at the first
bench in the greenhouse and also contained our pots that were spread out evenly in four
rows. Our pots stayed in the greenhouse for about five weeks, captured as much sunlight
as they could, and got their water source from sprinklers that automatically came on
twice a day.

Data Acquisition

After the duration of growth of the plants, our lab returned to the Greenhouse 2 to collect
data. The total number of plants, number of stems, average length (cm) of 5 randomly
chosen stems, weight of stems (g), the total number of leaves, weight (g) of the leaves,
and the weight (g) of the roots in each pot were measured.

The plants were handled very carefully when calculating measurements. The majority of
the plants was extremely fragile and had to be handled with a gentle hand. The plants
were removed from each pot by squeezing the sides of the pot to loosen the dirt. Once the
plant was ejected from each pot, we removed excess dirt off the tips of the roots while
still being careful not to rip the root tips off. A bucked of water was placed next to our
station to help the removal of the dirt off the root tips. Once the cluster of plants was out
of each pot, we counted the total number of plants for each pot, making sure to focus on
one pot at a time. Each leaf for each plant was cut off at the base, and weighed. Michelle
was recording all of the data while the rest of the group members worked in an assembly
line. The underground biomass was removed, weighed, and recorded. The stems were
removed, weighed, and recorded. For each pot, five randomly selected stems were
selected and measured. An average of all five measurements was recorded. This data was
recorded for all 24 pots. Data was recorded, and then shared amongst group members
after our station was cleaned up.

Data Analysis

A single and two factor ANOVA test was run on both species to compare their
mechanisms of survival and rate of success. This comparison was carried out using excel.
Specifically, the single factor tests were used to identify distinct differences between the
underground biomass and the above ground biomass of the two different species in pots
with different levels of density. Both of these tests also demonstrated significant
differences in the mechanisms and successes among density levels when under the
influence of intraspecific competition. The two factor ANOVA tests were used on the pots
that contained mixed species in order to quantify the effects of two species within the
same pot under different levels of densities. These results made clear whether or not there
was a distinct difference in the mechanisms and successes among density levels when
under the influence of interspecific competition.

The p-values yielded by the single and double factor ANOVA tests allowed us to say
whether there was a compelling difference in biomass in biomass between high density
and low density pots. This difference was only significant if the p-value was smaller than
0.05. These values specifically tell us if an effect of density or intercommunication is
present in the three different species levels of pots.

Results

In order to compare the intraspecific competition between collard plants and radish plants
in high and low density pots, the average root biomass was measured. For most plants,
water and nutrients are limiting resources that must be absorbed from their soil. In table
1, we see a p-value that is less than 0.05. From this data, we can conclude that a
significant difference is present in root biomass of radish plants in high and low-density
pots. From this data, we cannot conclude which exact pot contained a greater root
biomass. However, in Figure 2 we see that radish plant species had an overall greater root
biomass than the collard species. In table 2, we see a p-value that is greater than 0.05.
From this data, we can conclude that a significant different is not present in root biomass
of collard plants in high and low density pots.

Table 1. Single factor ANOVA test performed on the average root biomass of radish
plants in high and low density single species pots. The average root biomass is measured
in grams.

Anova: Single Factor

SUMMARY
Groups Count Sum Average Variance
Low 16 9.4025 0.583906 0.148885
High 16 2.369849 0.131866 0.016005

ANOVA
Source of Variation SS df MS F P-value F crit
Between Groups 1.624726 1 1.534726 17.67326 2.17E-04 4.160877
Within Groups 2.663346 30 0.082445

Total 4.308072 31

Table 2. Single factor ANOVA test performed on the average root biomass of collard
plants in high and low density single species pots.

Anova: Single Factor

SUMMARY
Groups Count Sum Average Variance
Low 16 5.080131 0.328133 0.201259
High 16 6.266865 0.381054 0.540466
 ANOVA
Source of Variation SS df MS F P-value F crit
Between Groups 0.04264 1 0.03254 0.126288 7.25E-01 4.160877
Within Groups 10.96438 30 0.375813

Total 11.02692 31

Figure 1 shows the results from the two single factor ANOVA tests more explicitly. A
greater difference between the high and low-density pots of the radish plants is shown.
The collard plants do not show a difference in density levels, which coincides with our
generated p-value. In the radish plants, the low-density pots had the higher average root
weight.
Average Root Weight

Radish
Collard

Low High
Plant Species

Figure 1. Density and species effects on root biomass in pots with single species. The bar
graph signifies underground biomass of both collard and radish plants in low and high-
density pots. High density represents pots with 64 plants while low density represents
pots with only 8 plants.

After observing the different in below ground biomass among the collard and radish
plants, it is vital that we observe the differences in total biomass between the two plants
as well. In table 3, we see a p-value that is smaller than 0.05. This value shows a
significant difference in the total biomass of the radish plants in both high and low
density pots. In table 4, we see a p-value that is also smaller than 0.05. Here, we are also
able to say that there is a significant difference in the total biomass of the collard plants in
both high and low density pots. Figure 2 tells us exactly which pots had a higher biomass.
The high-density pots had a greater biomass in both radish and collard plants.
Table 3. Single factor ANOVA test performed on the average above ground biomass of
radish plants in high and low density single species pots. The average biomass is
measured in grams.

Anova: Single Factor

SUMMARY
Groups Count Sum Average Variance
Low 16 194.5445 12.16965 24.02981
High 16 369.1223 23.0602 135.575

ANOVA
Source of Variation SS df MS F P-value Fc
Between Groups 952.3186 1 952.3097 11.82625 2.67E-03 4.1
Within Groups 2395.623 30 79.84742

Total 3348.032 31

Table 4. Single factor ANOVA test performed on the average above ground biomass of
collard plants in high and low density single species pots. The average biomass is
measured in grams.

Anova: Single Factor

SUMMARY
Groups Count Sum Average Variance
Low 16 39.66019 2.468137 2.329428
High 16 130.3804 8.139398 56.91376

ANOVA
Source of Variation SS df MS F P-value Fc
Between Groups 257.3156 1 257.3156 8.58642 5.15E-03 4.1
Within Groups 888.6378 30 29.61159

Total 1145.935 31
 Average Total Biomass

radish
collard

Low High
Plant Species

Figure 2. This bar graph shows the species and density effect on biomass that is above
ground in pots with single species. This graph also compares the average total biomass
calculated among radish and collard species in high and low-density pots. This graph
displays a higher average biomass in high-density pots in both collards and radishes.

Interspecific Competition Outcome:

Consideration must be given to the mixed species pots as well. In table 5, we see a two-
factor ANOVA test for the root biomass of the mixed species pots. This test yielded three
different p-values that must be taken into consideration. The p-value in relation to species
is greater than 0.05: this tells us that there is no underlying difference between species.
Radish and collards are the same regardless of the density. The p-value in relation to
density is less than 0.05: this tells us that there is a significant difference between the high
and low-density pots of mixed species. The p-value in relation to interaction is
approximately equal to 0.05: this tells us that there is no correlation between high and
low-density pots and type of species.

Table 5. Two factor ANOVA test performed on the average root biomass of collard and
radish plants in high and low density mixed species pots. The average biomass is
measured in grams.

Anova: Two-Factor with Replication

SUMMARY low high Total
Radish Mix
Count 16 16 32
Sum 17.48833 2.264469 19.7628
Average 1.083646 0.132154 0.5179
Variance 2.338807 0.017548 1.474823

Collard Mix
Count 16 16 32
Sum 4.738332 2.626286 7.465619
Average 0.296156 0.170293 0.232269
Variance 0.180732 0.155284 0.156766

Total
Count 32 32
Sum 22.22667 5.000654
Average 0.684896 0.155274
Variance 1.383743 0.074366

ANOVA
Source of Variation SS df MS F P-value F crit
Species 2.367053 1 2.367053 3.513679 6.57E-02 4.001181
Density 4.641833 1 4.641823 6.890374 0.010977 4.001181
Interaction 2.727377 1 2.727277 4.048548 0.048705 4.001181
Within 40.41007 60 0.673668
Total 50.14632 63
Biomass (g)

radish
collard

low high
Plant Species
 Figure 3. Density and species effect on the root biomass in the mixed species pots. The
radish biomass among the high and low density pots yields a notable difference; however,
this same trend is not seen in the collard species pots.

Consideration must be given to the total biomass in the mixed species pots as well. In
table 6, we see a two-factor ANOVA test for the total biomass of the mixed species pots.
This test yielded three different p-values that must be taken into account. The p-value in
relation to species is less than 0.05: this tells us that there is an underlying difference
between species. The p-value in relation to density is greater than 0.05: this tells us that
there is not a significant difference between the high and low-density pots of mixed
species. The p-value in relation to interaction is also greater than 0.05: this tells us that
there is no correlation between high and low-density pots and type of species.

Table 6. Two factor ANOVA test performed on the average total biomass of collard and
radish plants in high and low density mixed species pots. The average biomass is
measured in grams.

Anova: Two-Factor with Replication

SUMMARY low high Total

Radish Mix
Count 16 16 32
Sum 155.1822 203.2058 358.499
Average 9.698745 12.70655 11.20276
Variance 38.23245 24.94371 32.90836

Collard Mix
Count 16 16 32
Sum 17.62 14.97662 32.68552
Average 1.1065 0.170293 1.021635
Variance 1.205015 1.347758 1.381999

Total
Count 32 32
Sum 172.9032 217.2804
Average 0.684896 6.81216
Variance 38.16348 48.51892

ANOVA
Source of Variation SS df MS F P-value F crit
Species 1658.992 1 1658.992 100.9687 1.83E-13 4.001181
Density 32.19452 1 32.13267 1.955622 0.151722 4.001181
Interaction 40.42855 1 40.55724 2.65488 0.133373 4.001181
Within 987.6003 60 16.40155
Total 2717.53 63
 Biomass (g)

radish
collard

low high
Plant Species

Figure 4. Density and species effect on above ground biomass in mixed species pots. The
radish biomass among the high and low density pots yields a notable difference; however,
this same trend is not seen in the collard species pots.

Discussion

Table 3 shows a p-value of less than 0.05; this yields a notable difference in the total
biomass of the radish plants in high and low density pots. In table 4, we note a p-value
that is also smaller than 0.05. This shows us that there is also a significant difference in
total biomass between the low and high density pots among collard plants. Just
referencing the p-values may lead one to think that my hypothesis of both species
producing larger plants in low density pots than in high density pots is correct. However,
when you take a look at Figure 1, you note that high density pots yielded more total
biomass than plants in the low density pots. Therefore, I must reject my hypothesis. In the
high density pots, perhaps a portion of the seeds planted did not germinate in time. This
may have allowed the seeds that did germinate more room to grow and prosper, which
allowed them overall better and more access to nutrients.

When looking at the two factor ANOVA tests, both p-values related to interaction were
larger than 0.05. This tells us that there is not a notable interaction between density and
species type. We can accept our second hypothesis of the radish plants outgrowing the
collard plants below ground due to radishes having a larger root area and potentially
blocking the collards from obtaining the nutrients they need. In figure 3, we see a large
difference in the underground biomass of the radish plants in the high and low density
pots; however, we don’t see this difference in the collard plants. In figure 4, we see a
similar correlation in the different species among density levels. The difference in total
biomass of the radishes and collards in low and high denisty pots is almost equal. For this
very reason, the collard plants could potentially have a limiting resource that was not
observed in this experiment. For example, the interspecific competition could have not
effected the collard plant’s limiting resources. These resources could have been out of the
range that the pots resided in for the duration of the experiment. Or perhaps the collard
and radish plants could not out do one another, and were both drained access to the
limiting resources.

Future experiments could possibly include testing light as a limiting resource. Light is
very versatile, widespread, and the source of the majority of growth on earth. Minerals
and nutrients within the soil that was provided to these plants could also be tested. The
rate of water absorbance by radishes and collards, in relation to rate of growth in each
plant would be a great study to compare the size of a plant to it’s rate of prosperousness. I
feel this was a great introductory experiment to plant competition and how plants
compete to utilize resources to the best of their ability. We must continue to study the
effects of plant competition and breakdown, for their preservation allows us to live and
breathe everyday.

Acknowledgments

I would like to take the time to thank my lab partners that helped me in
conducting this experiment; Christina Watson, Michelle Saleeby, and Alexandra Mele.
Christina helped our group place the soil in all 24 pots during preparation. Alexandra
helped out a tremendous amount by labeling the pots initially with our section number
and plant information. Alexandra also removed dirt from the roots during the second part
of experiment, cut off leaves, measured stems, and weighed the under and above ground
biomass of each pot. Michelle helped out by recording the data as Alexandra and I
worked on each high and low density pot. I would also like to thank my TA, Shanna, for
guiding me through this experiment and being patient with all of her students. Shanna has
been great about explaining plant competition/breakdown and how it relates to what we
are studying. Likewise, I would like to thank the biology department of the University of
South Carolina for allowing me to have the opportunity to take this lab and for
administering to me the proper materials needed to conduct this experiment in a timely
fashion. Lastly, I would like to thank the owner of the Greenhouse 2 for allowing our
section to utilize space to carry out this experiment.
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