Heaney - Pisarski 1

Introduction
The farming industry accounts for over two hundred billion dollars of the United States
economy (Melillo). With global warming sweeping the nation, the growing season is severely
affected. People tend to focus on the here and now. The problem is that, once global warming is
something that most people can feel in the course of their daily lives, it will be too late to prevent
much larger, potentially catastrophic changes. says Elizabeth Kolbert, global warming
researcher. In a nutshell, global warming refers to the average rise in temperature of the Earths
atmosphere and oceans. This is caused by an abnormal amount of greenhouse gasses produced in
the form of smog and other air pollutants that damage our atmosphere and cause the suns heat to
intensify("Seriously, Just What Exactly Is Global Warming."). Climate changing, in particularly
heating up, droughts are popping everywhere scientists look ("How Global Warming
Works." ).This makes growing any type of crop harder than it should be.
With the attempt to speed up germination time citric acid solutions will be added to
radish seeds under varying temperatures. Varying amounts of citric acid (crystalline powder
form) will be added to water to create a solution that will be evenly distributed to the seeds. The
effect of temperature comes into play by placing the different concentrations of citric acid
solution and seeds into varying incubators creating constant growing temperatures.
The process of germination: the seed contains an immature plant (embryo) that resembles
an adult plant, complete with leaves and a root. The seed's leaves are called the cotyledons, seeds
that contain one embryonic leaf are known as monocotyledonous or monocots, whereas seeds
with two embryonic leaves are termed as dicotyledonous or dicots. The food found in the seed
which nourishes the embryonic seedling during its early stages of development, is known as
endosperm. With all the nourishment, the embryo becomes too large, and bursts open through the

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seed coat, in search of light to start its process of photosynthesis, and thus, the growing plant
emerges. During the same time, even the roots sprout and head down in search of more food
from the soil. Both the root and plant shoot move downwards and upwards, simultaneously and
respectively. In no time then, the seedling will force its way through the soil (Rajeev).
Radish seeds were chosen as the test subject mainly because of time restraints. They
germinate in 4-7 days naturally so results will show quicker than a seed that takes 2-4 weeks to
germinate. Citric acid, a solution with a 6% concentration will remove hard water stains from
glass without scrubbing. At room temperature, citric acid is a white crystalline powder; sold from
cookeries or brewing supply shops. (Ellis), commercially produced citric acid is mainly used for
preserving soft fruit drinks, and in home wine making. The citric acid will speed up the
germination time. This happens when the citric acid solution is added. It serves as the water that
any person would water their newly planted seed with. When the nourishment fills up the
embryo and pops the seed coat germination begins. With the added citric acid, the water goes in
as nourishments but the seed wants the citric acid out, making the embryo act as its filled up
and then bursting the seed coat faster than normal. The temperature effect is a naturally occurring
factor, flowing back to the issue of global warming. By monitoring the temperatures data will be
collected and analyzed. With rising temperature across the globe it was important that a high
temperature was tested. Originally in pretrials, even higher numbers were used but tests proved
that they were too high and were frying the seeds. The interaction of the two under the correct
circumstances should create a surge in germination time, resulting in a quicker growing crop.

Problem Statement

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Problem:
The effect citric acid and incubation temperature on the germination of Cherry Belle
radish seeds.
Hypothesis:
The lowest pH of citric acid and the highest temperature will help the Cherry Belle radish
to germinate the fastest.
Data Measured:
The independent variable are the amounts of citric acid and the temperature ( C) that are
being applied to our radish seeds. The dependent variable is the reaction of the Cherry Belle
radish seeds to the independent variable, affecting their germination. The growth rate of the
germination is going to be measured in millimeters (mm). The time is going to be measured in
days. A Cherry Belle radish seed is said to germinate in around 4-6 days. By using different
temperatures and different amounts of citric acid, it should increase the speed of germination
which will cause the plant to grow quicker.

Experimental Design
Materials:

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2 Raphanus Sativus seed packets with 100

seeds per packet
3 Imperial 3 lab line Incubators
900 grams citric acid powder
1 Paper towel roll
900 mL distilled water
1 Standard ruler with metrics
1 Permanent marker

(25) 616.51x14.92 cm plastic bags

(1) 100 mL graduated cylinder
(1) 2 mL dropper
1 Carolina analytical balance
(3) 250 mL beakers
1 Pencil
(1) 21x27 cm recording notebook

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Procedure:
Pre-Preparation:
1.

Label each bag for easy identification (name, date, treatment and group (ex. +,+).

2.

Rip off five paper towel sheets and place on clean working surface.

3.

Open one package of the radish seeds and place five radish seeds on each paper towel
spread equally apart.

4.

Fold the paper towel in thirds so that the seeds are enclosed with paper towel on each
side.

Figure 1. Diagram of the Setup of Radish Seeds On the Paper Towel.

Preparation of Solutions:
5.

Using the balance weigh out 0.1 grams of citric acid.

6.

Using the Graduated cylinder, fill to 100 mL.

7.

Make the first treatment (low) by combining the citric acid with the water inside one of
the beaker.

8.

Make the second treatment (standard) by using the same procedures 5-7 but instead of 0.1

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grams of citric acid use 0.2 grams.

9.

Make the third mixture (high) using procedures 5-7 instead using 0.3 grams of citric acid.

Incubation Samples:
8.

Taking a dropper add the 20 mL of the low mixture to the first five seeds previously
placed on the paper towel, moisten enough that it is clearly wet but not dripping.

9.

Repeat step 8 with the standard mixture and the high mixture.

10.

(Low) Gently place the paper towel with the five seeds and treatment in the bag labeled
(-,-) Place in the incubator with the lowest temperature (23 C).

11.

(Low) Gently place a second paper towel with five seeds and treatment in the bag labeled
(-,+) Place in the incubator with the highest temperature (36 C).

10.

(Standard) Gently place the third paper towel with five seeds and treatment in the bad
labeled (s) Place in the incubator with the standard temperature (30 C).

11.

(High) Gently place the fourth paper towel with five seeds and treatment in the bag
labeled (+,-) Place in the incubator with the low temperature (23 C)

12.

(High) Gently place the fifth paper towel with five seeds and treatment in the bad labeled
(+,+) Place in the incubator with the high temperature (36 C).

13.

Leave the seeds in the incubator for two days.

14.

Remove paper towel from bag and unwrap being careful not to knock off any seeds or
tamper with the results.

Table 1
Data Collection
First Day

Trial

Second Day
Amount of
Germinated
Seeds

Trial

(+,+)

(+,+)

(-,-)

(-,-)

Amount of
Germinated
Seeds

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Standard

Standard

(+,-)

(+,-)

(-,+)

(-,+)

Table 1 is used for the collection of data for each trial.

15.

Record data by observing what seeds germinated and how much growth occurred.

16.

Repeats for 5 trials; one trial every 2 days. Repeat all of the steps and record data.

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Data and Observations

Data:
Table 2
Design of Experiment Values
Factors

(-) Values

Standard

(+) Values

Citric Acid
(grams)

1/10

2/10

3/10

Temperature
(C)

23

29

36

Table 2 shows the high, low, and standard values for the different factors. The
values were chosen based on prior research and testing. Originally starting with 36C as
the low noticing that this produced germination but anything higher resulted in wilting
and frying of the seeds. The citric acid levels were also found by researching pH levels
and testing water pH with varying amounts of citric acid added. After testing, the results
leaned more to putting the least amount of citric acid possible seeing the differences
made. This is why 1/10 grams 2/10 grams 3/10 grams were chosen, as well as why 23 C
29 C 36 C were chosen.

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Table 3
Germination Data
Amount of Seeds Germinated
Citric Acid/Variable 1 Temperature/Variable 2

DOE
(-,-)

(-,+)

Standard

(+,-)

(+,+)

Standard

4 1/3

3 1/3

4 1/3

4 1/3

4 1/2

Average

Table 23shows the amount of seeds that germinated in each of the three trials.
Also showing the average amounts of seeds germinated per trial type.

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Observations:
Table 4
Observations
Trial
DOE

Standard

Seed germinations began splitting from dark seed coating

(-,+)

Trial went as expected

(+,+)

Small white fuzz appeared on 2 seeds, slight smell

(+,-)

Trial went as expected

(-,-)

Trial went as expected

seeds outer layer turned a lighter brown in color but still all
germinated
Seeds outer layer turned a lighter brown in color

(-,+)

The 1 seed that did germinate was small, just starting

(+,+)

Trial went as expected, slight smell

(+,-)

Seeds outer layer turned a lighter brown in color

Standard

Standard

(-,-)

Trial went as expected

seeds outer layer turned a lighter brown in color but still all
germinated
Seeds outer layer turned a lighter brown in color

(-,+)

Germination rate larger than normal

(+,+)

Mold grew on paper towel and seeds, slight smell

(+,-)

Trial went as expected

Standard

Trial went as expected

Standard

Trial went as expected

(-,-)

Standard

Observations

Trial Type

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Table 4 shows observations made after each trial took place. Looking at results for
anything that could be affecting the data.

(-,
+)
Figure 2. Trial as Expected
Figure 1shows an example of when the trial went as expected. Some or all of the
seeds germinated at a reasonable pace. This trial being (-, +) . Most standard tests also
came out with the trial going as expected. Almost every time 4-5 seeds germinated.

(+,
+) 3. White Fuzz Appeared
Figure
Figure 2. Shows when the white fuzz appeared in trial (+, +). The seeds shown on
the far left feature a fine coating. Another sign of defect was the mold growing on the
paper towel of a few of the trials performed.

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Data and Analysis

After prior research, it was decided to perform a series of trial runs on the effects
of citric acid and temperature on the germination of Raphanus Sativus radish seeds.
Table 5
Design of Experiment Values
(-)
Factors
Values Standard
Citric Acid
(grams per
100 mL)
1/10
2/10
Temperature
(C)
23
29

(+)
Values
3/10
36

Table 5 shows that factors were chosen to see how they affected the germination
of radish seeds. The amount of Citric Acid (grams) and temperature (C) were the two
factors. The table shows the high amount, low amount, and standard amount for each
factor. The values for Citric acid (grams per 100 mL) are as follows: 1/10 (low), 2/10
(standard) and 3/10 (high). The values for temperature (C) are 23 (low), 29 (standard)
and 36 (high).
Table 6
Averages
Trials

First
Run

Secon
d Run

Third
Run

Averages

Citric Acid Temperature

(+)
(-)
(+)

(+)
(-)
(-)

2
6
2

0
4
5

1
4
4

1
4 1/3
4 1/3

(-)

(+)

3 1/3

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Table 6 shows the amounts of the seeds germinated and each DOE. It also shows
the averages of each two factors for all three trials. The grand average is all four averages
added together and divided by four. Our grand average is 3.25 seeds germinated.

Table 7
Effect of Citric Acid
Citric Acid
+
4 1/3
1
3 1/3
4 1/3
Avg. 3.83333 Avg. 2.66667

6
4
Seeds Germinated

2
0
-1

Citric Acid (grams per 100 mL)

Figure 4. Effect of Citric Acid

Table 3 shows the amount of seeds that germinated when citric acid was applied.
It shows both the high and low amounts and the averages for both. When the low amount
was applied, an average of 3.8334 seeds were germinated. When the high amount was
applied, an average of 2 seeds germinated
Figure 4 shows the effect of citric acid. The effect is found from subtracting the low
amount (3.8333) from the high amount (2 2/3). The effect of Citric Acid on the
germination of seeds is -1.1667. This means that as citric acid increases, the seeds
decrease by 1.1667 seeds.

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Table 8
Effects of Temperature
Temperature
+
4 1/3
1
4 1/3
3 1/3
Avg.
4.33333 Avg.2.166667

5
4
3
Seeds Germinated 2
1
0
-1

1
Temperature

Figure 5. Effect of Temperature

Table 8 shows how many seeds germinated on average with the high and low
temperatures. When the high temperature was applied, an average of 4 seeds
germinated. When the low temperature was applied, an average of 2.1667 seeds
germinated. Figure 5 shows the effect of temperature on the germination of radish seeds.
This is found from subtracting the low amount (4 ) from the high amount (2.1667). The
effect is -2.167. This means that as the temperature increases, the seeds germination

decreases by 2.167.

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Table 5 is a table comparing the total averages between Citric Acid and Temperature when
adding them together in the experiment. Figure 3 shows the interaction effect of the citric acid
and temperature. The solid line segment represents the high amount for citric acid; the dashed
line segment represents the low amount for citric acid. The slope of the solid line segment is -5/3
and the slope of the dashed line segment is -1/2. By subtracting the low (dotted segment) from
the high (solid segment) you will get the interaction effect. The interaction effect of the two
factors is -1.1667.
In figure 2 when citric is held high it allows 2 2/3 seeds. If looking at the solid segment in
figure 4, citric acid only allows 1 seed. This represents the high citric acid with the interaction of
temperature. When temperature is held low, it allows 4 seeds to germinate. This is higher than
2 2/3 seeds. When temperature is low, the seeds germinate much better; when it is held high, the
seeds barely germinate.
In figure 2, when citric acid is held low, 3.833 seeds are expected to germinate. In figure
4 the dashed line represents the low of citric acid with the interaction of temperature. When
temperature is held high, the low amount of citric acid germinates 4 seeds. When the
temperature is held high, the low amount of citric acid is 3 seeds. This means that when the
citric acid is held low, no matter what temperature, it is still higher than the average amount of
seeds expected.

Table 10
Standards
5

Standard (seeds germinated)

5
5
5

Table 6 shows the germination of the standard seeds (2/10 grams citric acid and 29 C).
The range was from 3 to 5 seeds which means the range of standards was 2 seeds.

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Standard
s

Figure 7. Standards Plot

Figure 7 shows how many seeds germinated when applying the standard amount. The
lowest amount was 3 seeds and the highest was 5 seeds. During the last trial the amount of
standard seeds decreased because the seeds began to mold, causing them to germinate less than
the other trials. The range of standards is 2 seeds and when doubled it is 4 seeds.

Y= 3.25 + noise
Figure 5. Parsimonious Prediction Equation
Figure 5 shows the parsimonious prediction equation. Similar to the prediction equation
except using only the significant effects. None of the effects were significant making it just Y=
3.25 + noise

Y= 3.25
Figure 6. Parsimonious Prediction Figure 6 shows the parsimonious prediction which
predicts 3.25 seeds will germinate. This shows if the experiment was done again about 3.25
seeds would germinate on average. Every time the experiment was done, similar results appeared

Heaney Pisarski 17

which shows that there wasnt much variation. Being that there was no significant effects, it is
not possible to plug any factors into the equation.

2.167
T

CT

Figure 8. Dot Plot of Effects

Figure 8 shows the effects of the Citric Acid (C) and, Temperature (T), and their
interaction (CT). When thinking about significant factors that affect any experiment, a dot plot of
effects can be made. These effect values can be compared to twice the range of standards to see
if there are any significant factors based on the data. The dotted lines represent the range of
standards doubled, which in this case has an absolute value of 4. If the effects are outside the
dotted lines, they are said as statistically significant. This is because the effect exceeds the
standard even when it is doubled, making it significant. None of the effects were significant in
this case.

Interpretation:

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After doing the experiment over three trials, the evidence suggests that temperature
played a more significant role. Although it wasnt a statistically significant factor as shown in
figure 6 of the DOE. Citric acid did affect the data in a positive way during the standard runs. In
the process of the experiment and previous research, the researchers have concluded that citric
acid will indeed help the growing process when used to a specific amount able to change the pH
level in the seed. When combining the high amount of citric acid with the high temperature, the
seeds began to dry out. On the other hand when combining the low amount of citric acid with the
high temperature, many seeds germinated. Evidence shows that temperature did play the most
important role.

Conclusion

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It was hypothesized that the seeds with the highest amount of citric acid would germinate
the least amount of seeds. This hypothesis has been accepted throughout the data collection. The
point of the experiment is to add citric acid to radish seeds in order to change the pH. When the
pH changes it would stunt the growth properties of the seed. Varying amounts of citric acid were
added to groups of 5 seeds, each set of seeds were put into different temperatures to see the
differences in the growth. After prior research the experiment was decided because it contributes
to the growing problem of global warming. With a shorter, and harder to work with growing
season the quicker society can come up with ways to improve, the better.
The results came to conclude that the seeds combined with the high temperature and the
high amount of citric acid dried out. The seeds with the low citric acid and the low temperature
germinated a range of 5 to 4 seeds. Seeds with low citric acid and high temperature germinated a
range of 5 to 2 seeds and seeds with high citric acid and low temperature germinated a range of 4
to 2 seeds. All of the standard trials usually came out as 5 seeds germinated but during one trial
of the experiment the seeds began to mold which prevented the growth of the seeds and the seeds
did not germinate. This may have been because we reused the bags and exposed them to air
which created the mold. Citric acid was supposed to help the germination of our seeds which it
did because the radish seeds were supposed to germinate in 4-6 days but by adding the citric
acid, the seeds began to germinate in as little as 1 day. Citric acid helps to inhibit the formation
of mold and other bacterial growth, an all-too-common occurrence when sprouting in a closed
area due to high temperatures and humidity. Citric acid is used to prevent mold so it could help
the seeds to grow when higher temperatures were being used. ("Citric Acid - 4 oz..") .Our
experiment compares to other research found from adding tomato juice to germinate seeds.

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Tomato juice contains acid and sugar just like citric acid and when added to seeds it speeds up
germination. (Koning, Ross).
After searching the internet for a very long time the researchers couldnt find anything
like the experiment that was being performed so it could not be compared to other peoples work
and findings. Design flaws were present in the process. Starting with the bags, the same bags
were used over and over a few times after each trial. After observing the runs mold was present
on the high temperature bags which queued us to throw them away and get news bags. This
could have tampered our results if the mold was constantly growing, just not made present.
Another flaw dealt with the temperature. The low temperature was at 23 degrees Celsius. This
was the room temperature so the seed bags were placed on a table in the room. This could have
produced inconsistent results because of the varying amount of sun, body movement in the room,
etc. No errors were made when following the procedures. Further research could be conducted to
improve the experiment on mass production growing.
This would benefit the world wide problem of world hunger, and working to improve
harvesting time. If mass production of crops is done the same way that garden or small scaled
growing is done. a major design flaw had to do with the amount of seeds chosen to conduct the
trials and the standards dot plot. 5 seeds were placed on each paper towel, seed germination
ranged from 1-5 seeds depending on the trial number. according to the standards dot plot, after
doubling the range of standards it came to be -8 - 8. looking back to the amount of seeds there
were only 5 each. there was no possible way for there to be more or less than 8 for the
significance value, so it was impossible for any effect to be significant.
The information from the experiment can further be used in the real world to help the rapidly
increasing issue of global warming. At this time no one really knows about global warming and

Heaney Pisarski 21

what exactly its doing to our globe. By speeding up the process of germination times, the
growing season doesn't necessarily extend, but it makes it possible for more crops to be grown in
the allotted time. Benefits include a faster produced crop. The data proves that the combination
of citric acid and a target temperature the growth rates speed up. Faster growing crops mean
faster harvesting. The process is rather simple in that farmers would only have to water the seeds
with the citric acid solution once during the initial plant of the seed. The positive results are
plentiful. Not only will more crops be made, giving out more food, but farmers will make more
money due to more crops to sell.

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Acknowledgements
We would like to give a huge thanks to Mrs. Gravel for everything. She has been nothing
but a positive source always giving out critical help. Understanding we are under a lot of
pressure and making us smile any time given the chance. We would also like to thank each other.
The entire process of choosing the topic and getting everything to work out took a lot out of us.
We tried many different approaches and after fail after fail, we got through it. We constantly
pushed each other to make mental due dates and kept each other going the entire time.

Heaney Pisarski 23

Works Cited
Ellis-Christensen, Tricia . "What is Citric Acid?." . O. Wallace, 1 Jan. 2003. Web. .
<http://www.wisegeek.org/what-is-citric-acid.htm >.
"How Global Warming Works.". HowStuffWorks, 15 May 2012. Web. 21 May 2014.
<http://www.google.com/url?q=http%3A%2F%2Fscience.howstuffworks.com
%2Fenvironmental%2Fgreen-science%2Fglobalwarming1.htm&sa=D&sntz=1&usg=AFQjCNFXocCuuzJdr1YPMN-uBJrDveUgeA>.
KOLBERT, ELIZABETH. "QUOTES ON GLOBAL WARMING.". N.p., 25 Apr. 2005.
Web. 21 May 2014. <http://www.google.com/url?q=http%3A%2F%2Fwww.notable-quotes.com
%2Fg%2Fglobal_warming_quotes.html&sa=D&sntz=1&usg=AFQjCNHuu6tr02pTBEeAlTs7xnDIvXTUw>.
Melillo, J.M. "Agriculture and Food Supply.". US Environmental Protection Agency, 9
Sept. 2013. Web. 21 May 2014. <http://www.google.com/url?q=http%3A%2F%2Fwww.epa.gov
%2Fclimatechange%2Fimpacts-adaptation
%2Fagriculture.html&sa=D&sntz=1&usg=AFQjCNEn22oORLmbwvFvIwFHK_V_gEIz9Q>.
Rajeev, Loveleena. "Seed Germination Process.". Buzzle, 27 Nov. 2009. Web. 21 May
2014.<http://www.google.com/url?q=http%3A%2F%2Fwww.buzzle.com%2Farticles%2Fsegerminationprocess.html&sa=D&sntz=1&usg=AFQjCNGeHGo11DgSLh8OT394bOr_TG_jhw>
"Seriously, Just What Exactly Is Global Warming.". N.p., 10 Mar. 2012. Web. 21 May
2014.<http://www.google.com/url?q=http%3A%2F
%2Fwww.epa.govhttp://www.google.com/url?q=http%3A%2F

Heaney Pisarski 24

%2Fwww.climateexchangeplc.com%2F2012%2F03%2F10%2Fseriously-just-what-exactly-isglobal-warming%2F&sa=D&sntz=1&usg=AFQjCNEyn7YR9fFcdqyQIKaMEnlCKgZdjA
%2Fclimatechange%2Fimpacts-adaptation
%2Fagriculture.html&sa=D&sntz=1&usg=AFQjCNEn22oORLmbwvFvIwFHK_V_gEIz9Q>.