Brittany Rockey

4/15/10

Section 26

The Effects of Gibberellin and Paclobutrazol Hormones on Pea Plants Pisum sativum

Introduction

A hormone is a molecule that produces a signal in one part of an organism and transports

it to another part where it binds to a specific receptor and elicits a response within the cell.

Major plant hormones are auxins, cytokinins, gibberellins, brassinosteroids, abscisic acid, and

ethylene. The hormones that were explored specifically in this lab were gibberellins and

paclobutrazol. Gibberellins effect stem elongation, fruit growth, and seed germination, and in

this lab, the focus was on stem elongation. Paclobutrazol is a growth retardant that inhibits

gibberellins.

This purpose of this experiment was to look at the effects of different hormones on

plants. A pea plant was used in this experiment because it grows relatively fast and therefore

growth changes are easier to measure. Gibberellic acid is responsible for stem elongation and

therefore it will increase the length between nodes (internode length). Paclobutrazol hormone is

the antagonist of gibberellins. Paclobutrazol will slow plant growth, thereby decreasing changes

in internode length. The goal of this experiment is to explore the effects of these two hormones

on pea plant growth over a two week time frame.

It is predicted that when the pea plants are exposed to gibberellins, the internode length

will increase and when they are exposed to Paclobutrazol, the internode length will decrease.

Also, a plant that receives a higher concentration would be expected to show a larger increase in

internode length changes than a plant that was exposed to a lower concentration of that hormone.

The null hypothesis is that the different hormone treatments do not affect plant growth, so there
is no difference between growth in the control and in the other experimental plants. The

alternative hypothesis is that the hormones do have an effect on plant growth, and therefore the

different exposures should cause some kind of changes between the two experimental groups of

plants.

Materials and Methods

In this experiment, the class was set up into small groups of 3-4 students. Each group

received 5 plants-1 control plant, 2 that would be exposed to gibberellins first, and 2 that would

be exposed to Paclobutrazol first. It was the students’ decision if they wanted to have a high or

low concentration of the hormone applied to the plants. The plant height and internode lengths

were measured on the first day, and after that, they were measured on the next two Mondays,

Wednesdays, and Thursdays. The experiment was conducted for two weeks total, however, after

the first week, the plants initially exposed to gibberellins were switched to paclobutrazol, and the

plants that were first exposed to paclobutrazol were exposed to gibberellins. The same

concentrations (high or low) that were applied in the first week were also applied in the second

week.

The plants were all kept in the greenhouse and the greenhouse workers watered the plants

and treated the plants with the appropriate hormones for consistency. Data was collected by

visiting the greenhouse and using a ruler to measure the internode lengths and the height of the

plants. If needed, yarn was used to measure the lengths of the plants that did not grow straight

and were difficult to measure. On each of the days, the number of nodes, the length between the

nodes, and the total height of the plants were measured.

To calculate new internode length between days 0 and day 7, the following equation was

used:
(sum NEW Int L on Day2 + sum NEW Int L on Day4 + sum NEW Int L on Day 7)
( Total # NEW Int L measured on Day2 + Day4 + Day7)

This equation can also be used to find the internode on the other days by substituting data

different days into the equation. 1 To get better results, instead of using independent data from

each group, a set of pooled data from many different groups was used. The purpose of doing this

was to try to eliminate some person mistakes in measurements that may have made and to get a

more representative set of data. Due to poor data, some of the data was removed. Some of the

data had missing measurements for some of the days, or there was a large variance between the

values on two successive days that did not seem plausible.

To analyze the data, graphs were used to visually look for comparisons and rough trends

between the data. To simplify the labels for the data, a high concentration is denoted with an

“H” and a low concentration with an “L.” Also, gibberellins labels are shortened to “gib” and

paclobutrazol labels to “pac.” A t-test was used between many different groups to show if there

was any significant data that could be extracted from the lab. In a t-test, a p-value less than .05

indicates that the null hypothesis can be rejected and that the data is significant. This also

implies that the alternative hypothesis can then be supported. If a p-value is greater than .05, the

null hypothesis fails to be rejected. This suggests that there is support for the null hypothesis,

and also means that the data has no significance.

Results

Table 1: T-test Results

Groups Day 0 Day 7 Day 14
Control and gibH/pacH 0.161005 0.249984 0.288601

Control and gibL/pacL 0.843966 0.030972 0.132604

Control and pacH/gibH 0.210791 0.679113 0.029784

Control and pacL/gibL 0.883166 0.743512 0.667883

gibL/pacL and gibH/pacH 0.0984 0.846758 0.002373

pacL/gibL and pacH/gibH 0.139883 0.906614 0.097545

Table 1 shows the t-test values between different groups. Values highlighted in red indicate that the p-value
was below .05. The abbreviation in the label indicates the treatment given in the first week and the second
abbreviation indicates the treatment given in the second week.

Table 2: T-Test Week Results

gibH week one and gibH week 0.007696
2
gibL week one and gibL week 0.258683
2

pacH week one and pacH 0.000934

week2

pacL week one and pacL week 0.165442

2

Table 2 compares data between 2 weeks. Again, data highlighted in red represent a p-value
below .05.
 Figure 1 (above) and Table 3 (below) show the internode lengths at weekly intervals of the different treatments. The pacH/gibH and
pacL/gibL treatments decreased in internode length between week 0-1 and increased in length from week 1-2. GibH/pacH decreased in
internode lengths initially, but there was little difference in internode length from week 1-2. GibL/pacL increased the first week and
decreased the second week. The control showed a slight, steady decrease in internode lengths over the 2 weeks.

Table 3: Average Internode Lengths

Week 0 Week 1 Week 2
Control 16.8 11.6 10.3
gibH/pacH 16.8 27.6 32.63
gibL/pacL 12.5 15.5 11.7
pacH/gibH 16.6 11.8 30.3
pacL/gibL 12.3 8.1 11.6

Table 4: Average Total Height

Week 0 Week 1 Week 2
Control 68 79.4 111.3
pacH/gibH 46.4 77.9 232.9
gibL/pacL 57.5 108.9 138.4
pacL/gibL 55.7 70.6 128.1
gibH/pacH 49.1 116.3 287.7

Figure 3 compares the average total heights of the plants from both weeks with error bars showing the standard error. The label on the x-axis
represents the first treatment given to the plants. All of the treatments and the control show an increase in height during both weeks.
GibH/pacH grew and
Figure 2 (above) the most
Tableof4 all the treatments
(below) show the and the two
average totallow concentrations
height of the plantstreatment
in weeklygrew to almostThe
increments. the same
heightheight. The pacH/L
of the control increased
treatments
slightly at agrew therate.
steady leastGibH/pacH
in the first week.
steadilyAfter 2 weeks,
increased overthethetreatments
2 weeks. exposed
PacL/gibL to higher concentrations
increased slightly fromofweek
the hormones wereincreased
0-1, and then taller than
those given the treatments
more dramatically with1-2.
from week lower hormone followed
PacH/gibH concentrations.
the same pattern, except the increase in height was more extreme. GibL/pacL
increased between weeks 0-1, and kept increasing between week 1-2, but the increase from week 1-2 was less extreme.
Discussion
 In Table 1, most of the p-values exceed .05, however, those highlighted in red (gibL/pacL

and gibH/pacH and the control and gibL/pacL) have p-values that are within the alpha level of

significance. This means that for every piece of data above .05, we fail to reject the null

hypothesis and conclude that there is no real difference between the two groups being compared,

which makes the data not statistically significant. For the other two values, whose p-value was

below .05, we can reject the null hypothesis and conclude that the data for the groups was

significantly different and that there was an effect on the plant from the hormone. Notice that for

the groups that did have significant results, the gibberellins was the first hormone used.

In Table 2, half of the data turned out to be statistically significant. The t-test for GibH

week 1 and week 2 and pacH week 1 and week2 both yielded a t-test with p-values<.05. This

means that we reject the null hypothesis and conclude that the results are statistically significant

and that there is a relationship between the growth from week 1 to week 2 in these two

treatments. In the low concentrations of the hormones, the p-value was greater than .05,

therefore we fail to reject the null hypothesis and conclude that there is no relationship between

the growths of the plants from week 1 to week 2. The t-test points out that the higher

concentrations of the hormone produced significant results, which implies that the hormone

concentration also plays a role in growth. In both Tables 1 and 2 we should have seen more

statistically significant data, especially in the groups that measured high/low concentrations.

In Figures 1-3, it is expected that when the plants are exposed to gibH/L from week 0-1,

the plant internode length and overall plant height will increase and when exposed to pacH/L

from week 1-2, the internode length and plant height will decrease. Likewise, when exposed to

pacH/L from week 0-1, the internode length/height should decrease and when exposed to gibH/L

in week 1-2, it should decrease. Also, higher concentrations of the hormone should lead to more

dramatic results. So, when exposed to pacH, there should be more of a decrease in internode
length/height than when exposed to pacL and when the plant is treated with gibH, growth should

increase more than it does when it is treated with gibL. The control should grow steadily

throughout the 2 weeks of the experiment.

In Figure 1 and Table 3, the pacH/gibH and pacL/gibL treatments decreased in length

from week 0-1 and increased from week 1-2 and the results from the higher concentration groups

had more dramatic changes than the results from the lower concentrations. These two treatments

turned out the way they were expected to. In the gibH/pacH treatment, the length decreased

initially when it should have increased, and then after the first week, the internode length stayed

fairly constant, where it should have decreased. GibL/pacL increased the first week in internode

length, and decreased the second week. The control showed a slight, steady decrease in

internode length over the 2 weeks, and it was hypothesized to have a slight, steady decrease over

the 2 weeks.

In Figure 2 and Table 4, the average total heights of the plants are shown in weekly

intervals. The height of the control increases slightly over the 2 weeks at a steady rate, which

was the same as the expected results. GibH/pacH growth also increased over the 2 weeks, but

the growth was more dramatic than the control’s growth. It was expected that during the first

week, with exposure to gibH, the plant should have grown, but during the second week, growth

should have slowed down or stopped. PacH/gibH height increased slightly during week1, and

then increased dramatically during the second week, which was the expected growth. Also,

pacL/gibL followed a similar pattern, except the growth during the gibberellins treatment was

not as dramatic of a change in height as with the higher concentration of gibberellins.

GibL/pacL increased more during week 1 or treatment, but then continued to increase a little

more in height during week 2 as well. The results from the total average heights of the plants

were typical of the expected results, with the exception of the growth of gibH/pacH. Also, the
results expected as far as concentrations received were also in accordance with the expected

values (except for gibH/pacH) where the higher concentrations of the treatments were expected

to experience greater changes in plant height and the lower concentrations were expected to

experience smaller changes in height over the 2 week period.

Figure 3 and Table 4 compares the height differences of the two treatments during the 2

weeks (The initial heights were also included so growth during the first week could be compared

to the plant’s beginning height.). All of the heights increase over the two week period, where the

height was expected to increase when exposed to gibberellins and decrease or stay the same

when exposed to paclobutrazol. As expected, the plants treated with the higher concentrations of

the hormones had the greatest changes in height, and ended up being taller than those treated

with lower concentrations. The two lower concentration plants grew to approximately the same

height, which suggests that the concentration of the hormone had some kind of role in the

growth. The gibH/L treatments grew more in the first week than did the pacH/L treatments and

in the second week the pacH/gibH and pacL/gibL both had dramatic growth. Most of the data in

the figure had outcomes that were expected, with one of the few differences from expectations

being that gibH/pacL experienced a lot of growth during the second week, when growth should

have been decreasing or slowing down because of the paclobutrazol.

Many other experimenters have also used pea plants as their experimental organisms. In

a 2008 study, there was a group who tested the effects of plant hormones in seedlings. In the

study, there were 4 different sets of seeds which were placed in gibberellic acid for 24 hours, and

there was a control group that was just placed in water. After 14 days, pea shoot length was

measured. In the end, 2 of the 4 groups showed statistically significant results. The researchers

believed that all of the groups should have shown differences in growth and attributed their

results to poor seed quality and premature seed harvesting. The researchers say that experiments
with premature harvesting of dwarf pea plant seedlings and the effects of gibberellic acid on

them are part of their current areas of research.2 Similar to the results found in the lab, the

experimenters found 2 of 4 groups statistically significant (see Table 2). The results found in

this lab may also be attributed to poor plant quality (as well as other reasons), and it was

hypothesized that all of the groups should have shown statistically significant data.

In another experiment involving banana plants, different dosages of paclobutrazol were

applied to the soil for 10 months and different measurements were taken, including: height,

internode length, circumference of the stem, and the number of leaves. At the end of the study,

there was a 26% average reduction of plant height. This study showed similar trends with the

laboratory exercise. However, the height actually decreased over the course of the study,

whereas in the lab the growth only slowed. In the study however, they reported that, “growth

reduction could be observed 17 days after application of the product,”3 so it may be that the

plants in the lab just were not exposed to the paclobutrazol long enough for it to have a large

effect on the plant’s growth.

The results of this experiment were fairly useful, with a few significant p-values, and

much more data that followed trends of the expected outcomes (these can be found in Figures 1-

3). As with any experiment, better results could result with a longer period of time used for

growth, which is obviously pointed out in the study above, where it took 2.5 weeks of one

treatment to see any changes in growth. More groups pooled together to make the final data set

may yield more significant data, since the data should have more of a normal shape and should

be less skewed due to fewer outliers. Also, if each individual had their own group of plants, the

data would be more useful because it would eliminate that extra factor of measurement error

when different people find the height of the plant. Another variable that was unavoidable, yet

may have affected the growth of the plants was the fact that some plants just do not experience
growth as much as other plants, or it is possible that throughout the course of the 2 weeks that the

plant became damaged and growth was affected because of that. Even if the experiment was

carried out for one more week, any trends in the results may have been clearer, so future

experiments could have 3 weeks of observations with two measurements taken per week so that

trends in the data may be more obvious.

This experiment and other similar experiments may be useful in further exploring the

effects of plant hormone and the way hormones work that are inside of plants. Since not much is

currently known, and research on plant hormones is a new and growing trend, further exploration

with plant hormones and plant growth may help us further understand how plants work.

Research in this area may also help suggest ways that plants can be

manipulated/changed/selected to be more useful agriculturally or recreationally. Having plants

that can grow taller or shorter or just grow in different ways in general may be able to support

more edible items and therefore may give off more products. Also, this alteration in growth may

be something that people are interested in having around their homes just for aesthetic reasons.

The study of plant hormones and growth could be beneficial to humans in many ways.
References

1.Nelson K, and Burpee D (2010) Control of Plant Hormones and Tropisms. (Department
of Biology, The Pennsylvania State University).

2.Baumgartner S, Shah D, Schaller J, Kämpfer U, Thurneysen A, Heusser P (2008)

Reproducibility of dwarf pea shoot growth stimulation by homeopathic potencies of

gibberellic acid. Complement Ther. Med. 2008;16(4):183-91.

3.Maia E, Siqueira DL, Salomão LC, Peternelli LA, Ventrella MC, Cavatte RP (2009)
Development of the banana plants 'Prata Anã' and 'FHIA-01' under the effect of

paclobutrazol applied on the soil. 81(2):257-63.