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Introduction

With the Earth’s population growing every day, one important issue to humans is food.

How can we efficiently produce food and get it to those who need it while still having enough

land for people to live? Most people around the world eat food that is grown; fruits, vegetables,

and grains. These types of foods need to be grown and prepared in specific conditions in order

to be healthily and good to eat. One such solution to this is “vertical farms”. Vertical farms are

large skyscraper-like buildings with greenhouse-like rooms that have racks with shelves

containing plants stacked on these plants (“Vertical Farms: From Vision to Reality”). These

vertical farms can control important factors of growth that would otherwise be uncontrollable in

the environment outside. Of course, before you starting to grow plants in vertical farms, one

must know the appropriate conditions for each plant for the quickest and best results. This

experiment offers a solution to how these ideal conditions can be found.

Raphanus sativus (radish) seeds were used in the experiment. They were chosen as

they are fast and easy to grow. For effect variables, pH of water and temperature were chosen.

This is because pH and temperature are environmental factors that cannot be changed easily in

nature, so they would definitely need to be controlled in vertical farming.

The first factor of pH is the measure of the concentration of hydrogen ions in a solution,

or of the acidity or alkalinity of a solution (Senese). It is measured on a scale of 0 to 14, with 0

being very acidic, 14 being very high alkalinity (very basic), and 7 being neutral. Most

experiments focusing of the effect of pH on plants talk about the pH of soil. Most plants grow

best in slightly acidic soil, normally something such as 5.5-6.5 pH. This has to do with plant

nutrients which must be dissolved for plants to absorb them (Galloway). The level of pH affects

the solubility of the minerals, so pH influences the level of nutrients a plant receives. The seeds

will not be grown in soil and will only be watered. Whether water pH will have the same effects

as soil pH is to be determined.
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The second factor is temperature. Temperature is the degree of heat present in an

object. Heat is the energy produced by the movement of molecules in a substance. The faster

they move the more heat energy it has (“What is Temperature?”). Radishes ideal temperature

for germination is from about 13 °C to 29 °C. Temperatures of 24 °C, 27 °C, and 30 °C were

used because it is a good representation of natural temperatures. There were also material

limitations that prevented us from using a wider range.

R. sativus is an edible root that is a part of the Brassicaceae family. They are very easy plants

to grow and care for. They take four days maximum to germinate and usually take 3-6 weeks to

fully grow. To measure how water pH and temperature do affect R. sativus seed germination, a

two-factor Design of Experiment (DOE). A DOE tests two factors in low, standard, and high

categories. For pH: low=6, standard=7, and high=8. For temperature: low=24 °C, standard=27

°C, and high=30 °C. The factors will be tested on separate groups seeds as so: (+,+), (+,-), (-,+),

(-,-), and (S,S). Then it will be recorded how many seeds of each group germinated out of 55

seeds. From this it can be determined how each variable alone affected the response along with

how the interaction of the variable affected it. Standard values can be used to determine the

variability of the data and what effect values were statistically significant or not. From all this it

can be determined what conditions of water pH and temperature is best for R. sativus.

Problem Statement

Problem:

How does the pH of water and the temperature of surrounding area affect the rate of Raphanus

sativus seed germination?


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Hypothesis:

If the Raphanus sativus seeds are watered with water at a pH level of 7 and are grown in

27 °C, then they will germinate the most out of 55 seeds.

Data Measured

We will be changing the pH of the water, measured by the pH scale, and the

temperature of the surrounding area, measured in degrees Celsius. The growth rate of radish

seeds is the response variable, which will be measured in how many seeds out of 55 germinate

after one day. Results will be measured using a two-factor DOE. This means the two factors will

be divided into low, standard, and high categories and will each be tested on separate groups

seeds as so: (+,+), (+,-), (-,+), (-,-), and (S,S). For pH: low=6, standard=7, and high=8. This is so

different groups of seeds can be exposed to acidic, neutral, and basic water. For temperature:

low=24 °C, standard=27 °C, and high=30 °C. This is so the seeds can be tested at different

temperatures that are not too far apart that might harm the seed.

Experimental Design

Materials:

Distilled water 1 gal. Citric Acid 15 fl oz


Paper towels 11x6 in. (10) Sodium bicarbonate 1 lb
Champion radish seeds (825) pH test strips
Incubators (2) Dropper 1 mm
Gallon size ziploc bags (5) Box 1 ft. long 1 ft. wide
Beaker 50 mL Tupperware dish 13 cm x 9 cm x 3 cm (3)
Beaker 500 mL
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Procedure:

1. Set incubators to 27 °C and 30 °C

2. Place 300 mL of water in each tupperware dish.

3. Preparation for pH

1. Using dropper, put 115 drops citric acid in one of the dishes of water, pH will be

6.

2. Put 0.2 grams of sodium bicarbonate in another dish of water, pH will be 8.

4. Pull off five sets of two pieces of paper towel.

5. Put 55 seeds in a 5x11 pattern on one piece of paper towel.

6. Fold the other piece of paper towel on top of the seeds.

7. For each set, place 20 mL of water into the 50 mL beaker.

8. Using the dropper, spread the water evenly throughout the paper towel. Use water with a
pH of 6 on two sets, a pH of 8 on two sets, and a pH of 7 on one set.

9. Place the seeds watered with water at 7 pH in the 27℃ incubator.

10. Place one group of seeds with 5 pH in a box (24 °C) and one in the 30 °C incubator.

11. Place one group of seeds with 9 pH in a box (24 °C) and one in the 30 °C incubator.

12. After one day pull out the seeds and record how many germinated.

Diagram:
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Figure 1: Experimental Setup

Figure 1 above represents the seeds laid out on the paper towel in an 11x5 pattern.

Data and Observations

Data

Table 1
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Variable Values
pH (pH scale) Temp (⁰C)

Low (-) 6 24

Standard 7 27

High (+) 8 30

Table 1 shows the assigned values for each of the variables. For pH, the standard level

was set to 7 because this is neutral pH, with 6 being acidic and 8 being basic. The standard of

27 °C for temperature was determined because this is a temperature where the seeds are not at

risk of dying. As detailed in the procedures, pH was changed with citric acid (acidic) and sodium

bicarbonate (basic), and temperature was controlled using incubators (apart from the 24 °C, for

this is room temperature).

Table 2
Number of Seeds Germinated
Number of Seeds Germinated After One Day (Out of 55)
Trial Number
(+,+) (-,+) (+,-) (-,-) Standard

Trial 1 48.00 47.00 18.00 19.00 54.00

Trial 2 53.00 44.00 28.00 17.00 51.00

Trial 3 50.00 49.00 19.00 16.00 51.00

Average 50.33 46.67 21.67 17.33 52.00

Table 2 shows how many Raphanus sativus seeds out of 55 germinated after one day. A

total of three DOE trials were done and the averages of them all were taken.

Table 3
Percent of Seeds Germinated
Percent of Seeds Germinated After One Day (Out of 55)
Trial Number
(+,+) (-,+) (+,-) (-,-) Standard
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Trial 1 87.27 85.45 32.73 34.55 98.18

Trial 2 96.36 80.00 50.91 30.91 92.73

Trial 3 90.91 89.09 34.55 29.09 92.73

Average 91.52 84.85 39.39 31.52 94.55

Table 3 shows the percentages of the seed groups that germinated throughout the

whole experiment along with average values.

Observations

Table 4
Observations
Date Observation

March 9, 2015 Some of the seeds did not germinate after a week of pre-trials. This could
(Pre-trials) mean that they are dead or dormant. If that is true then that could alter our
results for the real trials.

March 11, -When the seeds were placed on the paper towel they were evenly spaced
2015 - March but sometimes when the towel was put in the Ziploc bags the seeds spread
20, 2015 out and were not evenly spaced anymore.
(Happened -Some of the seeds were colored black and may have played a role in the
Throughout the amount of seeds germinated (around 1 to 3 black seed(s) per trial).
Whole -The bags that came out of the 30° C (+) incubator had water droplets
Experiment) forming on the top more than the other two temperatures.

March 13, -When the seeds were looked at to count after several days roots grew
2015 straight through the paper towel.
(Before We -When counting the seeds some were just shells while others were still
Recorded whole seeds. This means that it took more focus to count the seeds.
Data)

Table 4 shows all observations that were noticed while the experiment was going on.

These may have had an impact on the experiment.


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Figure 2. Seed setup example

Figure 2 shows the spacing of the seeds from the (+,+) first trial. As the seeds were

placed they were evenly spaced for the most part but when the paper towel was picked up to be

placed in or on the Ziploc Bag the seeds sometimes moved. Also the seeds that are circled are

examples of some of the black seeds observed.

Data Analysis and Interpretation

Table 5
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Variable Values
pH (pH scale) Temp (⁰C)

Low (-) 6 24

Standard 7 27

High (+) 8 30

Table 5 shows the assigned values for each of the variables. For pH, the standard level

was set to 7 because this is neutral pH, with 6 being acidic and 8 being basic. The standard of

27 °C for temperature was determined because this is a temperature where the seeds are not at

risk of dying. As detailed in the procedures, pH was changed with citric acid (acidic) and sodium

bicarbonate (basic), and temperature was controlled using incubators (apart from the 24 °C, for

this is room temperature).

Table 6
Averages
pH and Temp Level (+,+) (-,+) (+,-) (-,-)

Averages Number of 50.33 46.67 21.67 17.33


Seeds Germinated

Grand Average 34.00

Table 6 shows the average amount of seeds germinated out of 55 throughout the DOE

experiment, along with the grand average (the average of the averages) of 34 seeds.

Table 7
Effect of pH
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pH (pH scale)

8 6

21.67 46.67

50.33 17.33

Average 36 32

Figure 3. Effect of pH

Table 7 and Figure 3 show the effect of pH to be 4 seeds (36-32=4). With this

experiment being out of 55 seeds,

pH most likely did not play a large

role.

Table 8
Effect of Temperature
Temperature (⁰C)

30 ⁰C 24 ⁰C

50.33 17.33

46.67 21.67

Average 48.5 19.5

Figure 4. Effect of Temperature

Table 8 and Figure 4 show the effect of temperature is 29 seeds. (48.5-19.5=29). With

this experiment being out of 55 seeds, temperature most likely did play a large role.
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Table 9
Interaction Effect

6 (-) 8 (+)

pH

30⁰ C (+) Temp 46.67 50.33

24⁰ C (-) 17.33 21.67

Figure 5. Interaction Effect

Table 9 and Figure 5 show the interaction effect of pH and temperature to be -0.33

seeds (46.67-50.33/1-(-1)=-1.83 and 17.33-21.67/1-(-1)=-2.16, so -2.16-(-1.83)=-0.33). The

slopes of the line segments look to be parallel to each other, so there does not appear to be an

interaction.
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Figure 6. Graph of Standards

Figure 6 shows all the values of the standards trials that were collected. The range of the

standards was three, meaning there was not a lot of variability in the data. Although more

standard trials should have been done, the lack of variability in these values indicates that the

data was not random and has a reason for being the way it is.

Figure 7. Test of Significance

Figure 7 shows a dot plot of the three effect values. Any value larger than double the

range of standards, represented by the bars, is significant. The temperature is the only effect

outside of the bars, therefore it is the only significant value.

Y = 34 + 29/2*T + “noise”

Interpolated prediction of (-0.5,-0.5)

Y = 34 + 29/2*(-0.5) + “noise”
Y = 34 + (-7.25)+ “noise”
Y= 26.75 seeds
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Figure 8. Parsimonious Prediction Equation

Figure 8 shows what the data may look like if the experiment was run with a 6.5 pH level

and a 25.5 ⁰C temperature level.

Data Interpretation:

From this data it was found that temperature was our only significant factor. The effect of

pH was small because the different pH levels did not vary much at all due to material limitations.

Weirdly, from research more acidic pH was expected to yield more seeds, yet more basic pH

did. The interaction effect was almost zero but interestingly was negative.

Conclusion

The hypothesis formed for this experiment was if the Raphanus sativus seeds are

watered with water at a pH level of 7 and are grown in 27 °C, then they will germinate the most

out of 55 seeds. This hypothesis was proven to be true. The experiment planned to find out how

different levels of water pH and surrounding temperature affected the germination of R. sativus

seeds. A two-factor DOE was used to compare low, standard, and high values of the two

variables to see which had significant effects on the response. Two factors were each tested on

separate groups seeds as so: (+,+), (+,-), (-,+), (-,-), and (S,S). For pH the experiment used a

low of 6, a standard of 7, and a high of 8. For temperature a low of 24 °C, a standard of 27 °C,

and a high of 30 °C was used. Water pH was made 6 (acidic) from citric acid and 8 (basic) from

sodium bicarbonate. Temperature was controlled by incubators, apart from 24 °C as this is room

temperature.

The standard values germinated the most seeds, 50.33/55. From statistical analysis, it

was found that temperature was the only significant factor for R. sativus seed germination.

Temperature had an effect of 29 seeds, more than half the amount used per run (55/2=27.5).
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The 30 °C and the 27 °C seeds had averages in the 40’s and 50’s, while the 24 °C seeds had

averages in the 10’s and 20’s. This shows that higher temperatures are better for the seeds.

Seeds continually deteriorate and unless they germinate they will die ("Temperature and Seed

Germination"). Moisture content can influence the rate of seed deterioration, and as higher

temperatures hold more moisture, they will help the seeds germinate. This is supported by the

large amount of water drops that were observed in the 30 °C bags.

The other main effect, pH, did not have a significant effect on R. sativus seed

germination. It only had an effect of 4 seeds. Strangely, from research it was found that slightly

acidic pH was the best for seeds, yet a positive effect suggests that slightly basic pH is the best.

Since acidic pH is lower on the pH scale, a negative effect value would support the pH level of 6

as the best. However, as stated before in the Introduction, most research done on this focusses

on soil pH, not water pH. Since we used neutral or near neutral levels of pH, it makes sense that

it did not have much of an effect. If the experiment was run again with further apart pH levels, it

would most likely be evident whether acidic, neutral, or basic water is the best for plants.The

interaction effect was 0.33 seeds, almost zero. The most interesting thing about the interaction

was that it was negative. The pH and temperature must have had a slight canceling effect on

each other, causing it to be negative. Since it was almost zero, it was not close to being

significant.

There are some other articles on the effect of pH or Temperature, but a article that had

both on radish seeds could not be found. One article that was found used the effect that pH had

on lettuce and poppy seeds. another thing that was different is they used a wider range of pH

levels (3-11). They found that the poppy seeds did not germinate under 5 pH but there was

slight growth for lettuce at 4 pH. Then at a range of 11 pH the poppy did not grow but lettuce did

(Powers, Mariko). In another article It talks about the effect of temperature on seed germination.

It talks about how if the temperature is too high it will evaporate all the water leaving it dry which

could negatively affect the germination. They used different temperatures at different times to
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simulate the effect of growing outside. This was different from ours where we used the same

temperature to simulate indoor growing (Nicholas, Dove). Another source gave the estimate

right conditions for the fastest germination. They suggested that you use 55 °F to 85 °F (around

13 °C to 30 °C) and wait 3 to 4 days for the seed to emerge from the ground and a pH ranging

from 5.8 to 6.8(“Explore Cornell”). these numbers were close and played a role in the choosing

of the numbers used in this experiment.

While following the experimental design and the procedures, there were a few setbacks

and flaws which may have affected results. Firstly, the low effect of pH shows further apart pH

levels should have been used to get a better understanding of how it affects the seeds.

Originally, pH level of 5 and 9 were planned to be used, but this was not possible due to

material limitations on the range of measurable pH levels. Also, mostly during the beginning, but

throughout the whole experiment, some seeds were difficult to tell if they germinated or not.

Since the data from the germinated seeds was collected after only one day, there were some

seeds that were hard to tell if they had begun germination or have not yet started. A procedure

was developed to tell if the seeds had germinated. If the seeds had split significantly or the stem

had emerged, they were counted as germinated. Though the researchers feel they did a good

job of telling the germinated and ungerminated seeds apart, there is a possibility of some error.

There are many exciting implications to further research and understanding of this field.

To begin, watching how the plants grow after they germinate is very important. Germination

rates are important, but it is grown plants that gives people the food they need, so watching how

factors influence plants throughout their whole lives is needed to determine the true effects of a

factor. Logically, these tests should also be run on different types of plants, along with using

different factors such as sunlight, soil content, amount of watering, and more. Experimenting

with all possible factors on all types of plants is critical in determining the best environmental

conditions for each individual plant type. This experiment and the other experiment options we

proposed apply directly to the vertical farms mentioned in the Introduction. Since these types of
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tests determine the best conditions for each plant to grow, they are essential for the success of

vertical farms, which may be the future of how food is grown.

During this experiment the researchers found out that the temperature had a larger role

on the germination of the R. sativus seeds and that pH played a small role. The experiment was

a good start for a wide area of plant growth. Next time variables that are farther apart and

adding new ones can help figure out what are the ideal environmental settings are the best for

growing certain plants in a controlled environment.


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Work Cited

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Novoselovic. "Seed Age and pH of Water Solution Effects on Field Pea (Pisum

sativum L.) Germination." Notulae Botanicae Horti Agrobotanici Cluj-Napoca. 1 Jan.

2007. Web. 6 Feb. 2015.

<http://www.notulaebotanicae.ro/index.php/nbha/article/view/246/238>.

Department of Agriculture, University of Reading, Earley Gate, UK. "Temperature and Seed

Germination." National Center for Biotechnology Information. U.S. National Library of

Medicine, 1 Jan. 1988. Web. 6 Feb. 2015.

<http://www.ncbi.nlm.nih.gov/pubmed/3077854>.

"Germinating Seeds." University of California, 1 Jan. 2005. Web. 15 Mar. 2015.

<http://www.uri.edu/cels/ceoc/documents/germinatingSeeds.pdf>.

Leubner, Gerhard. "The Seed Biology Place - Seed Germination." The Seed Biology Place -

Seed Germination. Royal Holloway, University of London, 12 Mar. 2005. Web. 14

Mar. 2015. <http://www.seedbiology.de/germination.asp>.

Olshan, Kraus Michael. "Effect of Temperature on Seed Growth." Effect of Temperature on

Seed Growth. 16 Nov. 1998. Web. 15 Mar. 2015.

<http://web.horacemann.org/academics/science/expbio99/Olshan.htm>.

"Explore Cornell - Home Gardening - Vegetable Growing Guides - Growing Guide." Explore

Cornell - Home Gardening - Vegetable Growing Guides - Growing Guide. 14 June

2013. Web. 13 May 2015.

<http://www.gardening.cornell.edu/homegardening/scene341b.html>

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<http://coolcosmos.ipac.caltech.edu/cosmic_classroom/light_lessons/thermal/tempera

ture.html>

Senese, Fred. "What Is PH?" General Chemistry Online: FAQ: Acids and Bases:. 15 Feb.

2010. Web. 13 May 2015.

<http://antoine.frostburg.edu/chem/senese/101/acidbase/faq/what-is-pH.shtml>

"Vertical Farms: From Vision to Reality – State of the Planet." State of the Planet Vertical

Farms From Vision to Reality Comments. 17 Oct. 2011. Web. 13 May 2015.

<http://blogs.ei.columbia.edu/2011/10/13/vertical-farms-from-vision-to-reality/>.