Effects of Caffeine on Plant Growth Adrienne Fraser and Rob Forgeron EDUC5370 Science Methods Professor Marilyn Webster

r November 20, 2012

Adrienne & Rob General Concept:

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How does caffeine affect the growth of plants? Big Questions: How does ground coffee affect plant growth (height & leaf count) when mixed with the potting soil? (versus Control pot with only soil) Is it the caffeine that affects the growth or some other factor within the coffee? o Use Decaf coffee in another pot, and use crushed Caffeine Tablets in a third, record differences. Could the same effect be achieved by watering the plant with [cooled] brewed coffee (versus mixing it into the soil)? Will regular tea (orange pekoe) achieve a different result than those obtained using the previously mentioned methods? If the caffeine is providing some extra energy (accelerating growth), will a plant with coffee grounds in the soil grown in majority shade grow as much as a plant without coffee that is grown under sunlight? Independent Variables: Soil contents, Sunlight, Watered with ___, Time

Dependant Variables: Height (mm), Leaf Count (integers), general appearance/health

Pot Labels and their Contents: A Control (regular potting soil, grown in normal sunlight, water) B Regular Coffee (potting soil + coffee, normal sun, water) C Decaf coffee (potting soil + decaf coffee, normal sun, water) D Caffeine Tabs (potting soil + crushed caffeine tabs, normal sun, water) E Brewed Coffee (potting soil, normal sun, cooled brewed coffee) F Tea (potting soil + tea leaves, normal sun, water) G Coffee w/o sun (potting soil + coffee, shade, water) 1 = big seed/garbanzo, 2 = little seeds/flax [see Image 1]

Adrienne & Rob Introduction and Research:

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Caffeine is a natural substance which has been in use for many years now. Most commonly it is the reason that coffee is consumed all around the world. It is found in many different kinds of plants and is not exclusive to coffee in particular. In humans caffeine acts as a stimulant to our central nervous system and when used correctly wards off drowsiness and restores mental alertness. The effect of adding caffeine to the soil and its effect on plant growth is a subject that has been studied for quite some time. Some plants seem to benefit and grow faster when caffeine is added to the soil, while others seem to become stunted or grow slower. There are also some plants that are not affected at all by the presence of caffeine in the soil. There are many ways which caffeine can be introduced to the soil. You can administer caffeine by sprinkling grounded coffee over the soil, adding leftover coffee to the pot or watering with a caffeine solution made by dissolving a caffeine tablet in water. Since the grounded coffee is actually organic matter and will help in adding nutrients to the soil which could also potentially have an effect on plant growth. In nature worms that feed on the grounded coffee may be attracted and at the same time that they are feeding they can help to aerate the soil. Many gardeners are known to add grounded coffee to the soil in order to accelerate plant growth. However, it is not clear whether the improved growth is due to the caffeine or the presence of potassium and phosphorous in coffee. It is also possible that caffeine slows plant growth but the presence of potassium and phosphorous in coffee reverses this effect. Caffeine like many substances can be toxic to organic life in certain concentrations. Since we are stimulated by caffeine we hope to see if plant growth can also be stimulated which could result in accelerated growth of our seedlings.

Adrienne & Rob Materials:

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14 x 14oz. Solo Cups (7 holes poked in bottom of each with a pen) 14 Cups Potting Soil 2 Cups Regular Ground Coffee [Melitta Dark Roast] (B1&2 and G1&2) 1 Cup Decaf Ground Coffee [Tim Hortons] (1/2 Cup each in C1 and C2) 6 x Caffeine Tabs [crushed] (3 each in D1 and D2) 10 teabags [Regular Orange Pekoe Tea] (5 each in F1 and F2) 7 x Big Seeds (see Image 1) 28 x Little Seeds (see Image 1) Approx. 2 Cups Brewed Coffee (used to water E1 and E2) [ Approx. 12 Cups water (used to water remaining plants) [ Cup x 2 Pots x 3 days] Cup x 12 Pots x 3 days]

Procedure: 1. Before planting each Solo Cup was labelled with its own letter/number identifier (see Pot Labels), as well as a brief description of what was in that pot. Each cup had 7 evenly spaced holes poked in the bottom with a pen, for soil drainage. 2. 12 of the cups were placed on a table by Adriennes back door (sliding glass door) which receives sun from sunrise until approximately 1 or 2 pm. The remaining 2 (which are to receive shade) are on the counter next to the wall, under a cupboard, where there is very little, if any, natural sunlight. See images 1 and 2 for pot locations. 3. 1 cup of soil was added to each cup. 4. The additives being tested (coffee, decaf coffee, loose tea, caffeine tabs) were then added to their respective cups and mixed thoroughly into the soil. 5. 2 kinds of seeds were planted for each variable, and all the seeds were placed at a depth of approximately 1.5 cm. 6. Following planting each pot was watered with 1/3 cup of water, except pots E1 and E2 which were watered with 1/3 cup of cooled brewed coffee each. 7. The plants receiving sunlight were spaced evenly on a large cookie sheet and placed on the table next to the door. The plants being grown in the shade were placed on a plate on the kitchen counter.

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8. The plants were watered every 7 days (once a week) with either 1/3 cup of water or 1/3 cup of cooled brewed coffee. 9. The plants in the sunlight were rotated every second day to ensure they were receiving equal amounts of sun. 10. Each plant was measured daily using a tape measure marked with centimetres and millimetres (Table 1); the number of leaves on each plant was counted daily (Table 2); the plants were assessed using a Green Scale (Appendix A) daily (Table 3). 11. Photos were taken every 2-4 days; qualitative observations were recorded every 2-4 days, depending on the amount of change seen. Skills: This experiment made use of many scientific skills, such as: Measurement - soil, additives, water, plant height Data Organization and Analysis Graphing Quantitative and Qualitative Observation Basic Statistical Analysis growth rate, means, standard deviations Problem Solving how to test for our desired variables Critical Thinking analysis and possible explanations of qualitative observations, data, and results.

Adrienne & Rob Qualitative Observations:

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Day 1 [October 4] Today we planted all of the plants; see Procedure Day 3 [October 6] No sign of growth in any of the pots that we can distinguish. Day 5 [October 8] No sign of growth in any of the pots that we can distinguish. Day 6 [October 9] A tiny shoot has started growing in Control 2 (A2). Day 7 [October 10] F2 (Tea in Soil) has started growing. Its pretty tiny right now so it is difficult to even make out the colour. The control (A2) is growing fairly quickly. Day 10 [October 13] E2 (Watered with Brewed Coffee) has started growing. No 1 cups (garbanzo) have sprouted at all yet. All of the cups are starting to grow a fine layer of blue/grey mold on top of the soil (probably due to the moisture + sunlight + food elements in the soil) hopefully this wont affect our plants, but we may have to look at it closer later. Day 13 [October 16] C2 (Decaf in Soil) has started growing. Our control plant looks to be doing the best of all of them it is bright, the stalk is thick, and it is growing in very quickly with several leaves. Day 14 [October 17] G2 (Coffee in Soil, no sunlight) has started growing. It looks pretty scrawny and yellow-brown (see Table 3) and not like it is going to grow very well at all. Now that we are 2 weeks into the experiment there is still no sign of any of the 1 cups (garbanzo beans) sprouting this probably has more to do with the seed used than any of our variables. Day 17 [October 20] All of the plants that have sprouted seem to be growing very quickly, although some definitely look better than others. For instance A2, E2, and F2 are all fairly thick-stalked and growing straight, whereas G2 (in the shade) is very spindly and seems to be growing horizontally towards the light from the pot lights in my kitchen. Day 19 [October 22] It would appear that the garbanzo beans are not going to grow under any of the conditions we have provided. We will have to compare results for flax plants only. Day 23 [October 26] The control plant is still going strong it has 5 different leaves now, and is about 22 centimetres tall. The plant with tea in the soil is also growing very well it does not have as many leaves as A2, but it is similar in height and colour. G2

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(in the shade) is still growing in height, although it perhaps should be considered length for this plant as it is growing horizontally almost perfectly parallel to the counter. E2 (watered with brewed coffee) and C2 (decaf in soil) look fairly average and similar. Day 25 [October 28] There appears to be very little change in any of the plants now, except for slight height growth. After 25 days it is doubtful that any of the plants which have not sprouted yet are going to (especially given the layer of mold on top of the soil in all the cups). The plants that have grown all seem to have levelled off in relation to one another. The mold/smell in my kitchen is getting pretty bad so we have decided to terminate the experiment at this time. It would appear that none of our conditions improved the growth of the plant, although tea in the soil and watering with coffee also did not seem to do much damage. These results will be further analysed in the discussion and conclusion. Photos:

Image 1: seen on top are 5 of the small seeds (flax) planted in all pots labelled with a 2; the large seed (garbanzo) on the bottom was planted in all pots labelled with a 1.

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Image 2: Pots A1-F2 [on Day 1] which will receive sunshine

Image 3: Pots G1 and G2 [on Day 1] which will remain in the shade

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Image 4: Overhead shot of all of the plants in the sun (Day 7)

Image 5: All Pots (Day 9) note growth in A2 (bottom) and F2 (top)

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Image 6: Plants sprouted so far (Day 10) Left to Right A2 [Control], E2 [Watered with Coffee], F2 [Tea in Soil]

Image 7: Same plants as Image 6, shot from above to compare stalk thickness and colour.

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Image 8: A2 Day 10 Close-Up

Image 9: E2 Day 10 Close-Up

Image 10: F2 Day 10 Close-up

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Image 11: All sunlight plants Day 13

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Image 12: Height comparison shot of Plants sprouted thus far (Day 13)

Image 13: E2 Day 13

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Image 14: C2 first sprout (Day 13)

Image 15: Sickly G2 (Day 18)

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Image 16: Sunlight Plants Day 18

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Image 17: F2 Day 18

Image 18: A2 Day 18

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Image 19: E2 Day 18

Image 20: G2 Day 24

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Image 21: E2 Day 24

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Image 22: E2/F2 day 24

Image 23: A2 Day 24

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Image 24: C2 Day 24

Adrienne & Rob Data Tables:

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Table 1 Height of Each Plant (mm) DAY Pot A1 Pot A2 Pot B1 Pot B2 Pot C1 Pot C2 Pot D1 Pot D2 Pot E1 Pot E2 Pot F1 Pot F2 Pot G1 Pot G2
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25

0 0 0 0 0 0 0 0 0 0 0 0 0 0

0 0 0 0 0 0 0 0

0 23 0 0 0 0 0 0 0 21 0 34 0 0

0 39 0 0 0 0 0 0 0 27 0 58 0 0

0 56 0 0 0 6 0 0 0 43 0 72 0 0

0 108 0 0 0 60 0 0 0 95 0 114 0 60

0 122 0 0 0 76 0 0 0 102 0 120 0 76

0 130 0 0 0 83 0 0 0 114 0 127 0 90

0 145 0 0 0 90 0 0 0 136 0 145 0 111

0 168 0 0 0 97 0 0 0 142 0 159 0 120

0 193 0 0 0 104 0 0 0 157 0 194 0 123

0 221 0 0 0 109 0 0 0 170 0 217 0 138

0 238 0 0 0 111 0 0 0 188 0 237 0 145

0 262 0 0 0 113 0 0 0 195 0 255 0 156

0 0 0 0 2 3 3 5 14 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

87 98 99 0 0 0 23 0 0 0 53 0 81 0 8 0 0 0 37 0 0 0 72 0 97 0 23 0 0 0 52 0 0 0 81 0 111 0 41

0 0 0 0 0 0 0 0 11 0 0 0 0 0 0 0 0 0

0 0 0 0 0 4 7 9 21 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

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Table 2 Number of Leaves on Each Plant (in whole numbers)

DAY Pot A1 Pot A2 Pot B1 Pot B2 Pot C1 Pot C2 Pot D1 Pot D2 Pot E1 Pot E2 Pot F1 Pot F2 Pot G1 Pot G2

4 5

6 7 8 9

10

11

12

13

14

15

16

17

18

19

20

21

22

23

24

25

0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

0 1 0 0 0 0 0 0 0 1 0 1 0 0

0 1 0 0 0 0 0 0 0 1 0 1 0 0

0 2 0 0 0 1 0 0 0 1 0 1 0 0

0 3 0 0 0 1 0 0 0 1 0 1 0 1

0 3 0 0 0 1 0 0 0 1 0 1 0 1

0 3 0 0 0 1 0 0 0 2 0 1 0 1

0 4 0 0 0 1 0 0 0 2 0 1 0 2

0 4 0 0 0 1 0 0 0 2 0 1 0 2

0 4 0 0 0 1 0 0 0 2 0 1 0 2

0 4 0 0 0 1 0 0 0 2 0 1 0 2

0 4 0 0 0 1 0 0 0 2 0 2 0 3

0 5 0 0 0 1 0 0 0 2 0 2 0 3

0 5 0 0 0 1 0 0 0 2 0 2 0 4

0 5 0 0 0 1 0 0 0 3 0 2 0 4

0 5 0 0 0 1 0 0 0 3 0 2 0 4

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Table 3 General Appearance Rating on a scale from 1-6 (See Scale in Appendix A) DAY Pot A1 Pot A2 Pot B1 Pot B2 Pot C1 Pot C2 Pot D1 Pot D2 Pot E1 Pot E2 Pot F1 Pot F2 Pot G1 Pot G2
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25

0 0 0 0 0 0 0 0 0 0 0 0 0 0

0 0 0 0 0 0 0 0 0 0 0 0 0 5 5 5 5 5

0 5 0 0 0 0 0 0 0 2 0 2 0 0

0 5 0 0 0 0 0 0 0 2 0 2 0 0

0 5 0 0 0 3 0 0 0 2 0 3 0 0

0 5 0 0 0 2 0 0 0 2 0 3 0 2

0 5 0 0 0 2 0 0 0 3 0 3 0 2

0 5 0 0 0 2 0 0 0 3 0 3 0 2

0 5 0 0 0 2 0 0 0 3 0 3 0 2

0 5 0 0 0 2 0 0 0 3 0 3 0 1

0 5 0 0 0 1 0 0 0 3 0 3 0 1

0 5 0 0 0 1 0 0 0 3 0 4 0 1

0 5 0 0 0 2 0 0 0 3 0 4 0 1

0 5 0 0 0 2 0 0 0 3 0 4 0 1

0 5 0 0 0 2 0 0 0 3 0 4 0 1

0 5 0 0 0 2 0 0 0 3 0 3 0 1

0 5 0 0 0 2 0 0 0 3 0 3 0 1

0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 3 3 3 2

0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

Adrienne & Rob Graphs :

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Figure 1: Plant Growth Over Time

300

250

200

Height (mm)

Pot A2 - Control 150 Pot C2 - Decaf Pot E2 - Brewed Coffee Pot F2 - Tea 100 Pot G2 - Coffee in Shade

50

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25

Day

Figure 1 shows the relative growth rates for each of the plants. The average slope of each line (ie Growth Rate) is very similar, which shows that although some plants started growing later, their growth rates were all comparable.

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Figure 2: Leaf Count Over Time

Number of Leaves

4
3 2 1 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25

Pot A2 - Control Pot C2 - Decaf Pot E2 - Brewed Coffee Pot F2 - Tea Pot G2 - Coffee in Shade

Day
Figure 2 gives a better indication of the possible effect that caffeine had on the plants (except for the control). The control plant overall was the healthiest in every aspect; however comparing all of the experimental plants, those with more caffeine (ie Coffee in Shade, Brewed Coffee) grew more leaves than those with less caffeine (ie Tea, Decaf).

Figure 3: Green Scale Rating Over Time

Green Scale Rating (see Appendix A)
6 5 4 Pot A2 - Control 3 2 1 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 Pot C2 - Decaf Pot E2 - Brewed Coffee Pot F2 - Tea Pot G2 - Coffee in Shade

Day
Figure 3 shows the relative green scale rating of each plant, which can be used to determine whether the plant was healthy or not. This shows that the plant in the shade (G2) was the least healthy, and the control plant was the healthiest.

Adrienne & Rob Data Analysis and Calculations: Sample Growth Rate Calculation:

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Mean Growth Rate Calculation:

Mean Plant Height Calculation:

Adrienne & Rob Standard Deviation Calculation: Plant Growth Rate [G] (mm/day) A2 - Control C2 - Decaf E2 Brewed Coffee F2 - Tea G2 Coffee in Shade 13.1 8.69 13 13.42 13 0.858 -3.552 0.758 1.178 0.758 G - G

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(G - G)2 ([mm/day]2) 0.736 12.617 0.575 1.388 0.575 =

Final Height [h] (mm) 262 113 195 255 156

h - h (mm) 65.8 -83.2 -1.2 58.8 40.2

(h - h)2 (mm2) 4329.64 6922.24 1.44 3457.44 1616.04

(mm/day)

Sample Variance [Growth Rate] Standard Deviation [Growth Rate] Sample Variance [Height] Standard Deviation [Height] =

Plant

Growth Rate (mm/day)

Mean Growth Rate (mm/day)

Within How Many Standard Deviations of the Mean?

Height (mm)

Mean Height (mm)

Within How Many Standard Deviations from Mean?

A2 Control C2 - Decaf E2 Brewed Coffee F2 - Tea G2 Coffee in Shade

13.1 8.69 13 13.42 13

12.242 12.242 12.242 12.242 12.242

1 2 1 1 1

262 113 195 255 156

196.2 196.2 196.2 196.2 196.2

2 2 1 2 1

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Looking at these calculations we can see that all of the plants except for C2 had growth rates that were not statistically significantly different. Even C2 was within two standard deviations of the mean, showing that although it was less consistent than the others, it was still not greatly significantly different. A greater difference can be seen in the final heights of the plants. E2 and G2 were the most statistically close to the mean, but the other 3 plants were all within two standard deviations. The differences here were more due to how long it took the seeds to sprout initially, and less due to their growth rate. After examining these statistics, it is impossible to conclude whether any of the substances had a significant effect on the growth of the flax plant. Since the control actually grew the best, it could be concluded that the substances all had negative effects, but further experimentation and much more data would be needed to substantiate these conclusions. Discussion: 1. Why did the garbanzo beans not sprout at all? It is unknown why the garbanzo bean did not sprout . Nothing definitive to be had in terms of research. 2. What were the weaknesses of the experimental design / implementation? Strengths? Weakness of experiment include not precise laboratory conditions, not enough trails for each seed , not enough variation in type of seeds. Growth time not long enough in duration. Details about specific nutrient and micorriza content of the soil used was not known. Strengths of experiment include the multiple ways used to observe and collect data from successful and unsuccessful trails. Small focus on two species to draw specific conclusions from. 3. How could the experiment be improved and/or extended for future use in the classroom? Experiment could be improved and extended by using an increased amount of different seed types to gather a more generalised sample of plant life and its reaction to caffeine. Also growth time could be extended to observe the impact of caffeine on the complete life cycle of our plants. 4. What assumptions were made that may have hindered the experiments success? Several assumptions were made during this experiment. All of them could have had unknown effects on the outcomes. These assumptions included: a. The regular and decaf coffees were identical except for their caffeine content.

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b. The amount of water (or brewed coffee) received was adequate but not too much. c. All plants placed by the window received equal amounts of sunlight. d. There were no differences in air temperature or other variables between the sunlight location and the shade location. e. Garbanzo and Flax seeds take the same amount of time to sprout and grow, and require similar growing environments. 5. What were possible sources of error in this experiment? Looking at the assumptions we made when designing the experiment, all could be perceived as sources of error. There was a bit too much variability in the environments that we did not measure or control for that may have had un-known effects on the plants growth. Other possible sources of error were measurement error (ie water, soil additives, plant height, etc); the subjectivity of the green scale measurements; unknown soil contents prior to beginning experiment. Conclusion: Unfortunately the results obtained during this experiment lead to no definitive conclusion regarding the effects of caffeine on plant growth. We can make tentative deductions, but there is not enough experimental data to conclusively respond to any of the questions this experiment attempted to answer. For example, since the plant with decaf coffee was the only experimental plant that did not contain caffeine, and since it had the lowest overall growth rate and final height, we can infer that the caffeine in the other plants did have some effect on the plant growth, and not other components of the additives. The most confusing result was the growth of the plant that contained ground coffee in the soil and was grown in the shade. The plant with identical starting components that was placed in the sun never sprouted at all, meaning we have no basis for comparison of the energy needed from the caffeine in the shade versus in the sun. After examining all of our assumptions and possible sources of error, it is clear that some extra variable controls should have been in place, as well as a bit more research into the proper way to sprout and grow flax and garbanzo plants before the experiment was started. Looking at the statistical data analysis there were no significant variations in height or growth rate between the different growing environments; the only conclusion that can be

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drawn at this point is that further data is needed in order to stabilize the data and find consistent trends in the effects of various sources of caffeine as additives in soil. Appendix A: Green Scale

Appendix B: Update Schedule Updates were provided on the following dates: Tuesday, September 18, 2012 (project topic proposal, in class) Monday, October 8, 2012 (via email) Tuesday, October 23, 2012 (in class) Tuesday, October 30, 2012 (via email) Sunday, November 11, 2012 (via email) Monday, November 19, 2012 (via email) Appendix C: Scientific Drawing Drawing an organism is a good learning aid because it makes you observe and accurately the form , proportion , and features of the organism. Page Headings: 1. Put your name on the top right of each page. 2. Put the Figure number on the bottom right of each page. Titles: Each drawing must have a title at the bottom of the page , centered neatly on the page, which includes as many of the following points as possible: 1. Binomial or scientific name of the organism. 2. Condition of the specimen 3.The portion of the specimen drawn 4. The View 5. Sex of the organism 6. Magnification of your drawing compared to the actual specimen

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Adrienne & Rob References:

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Dunn, D. (2012). The Effects of Caffeine on Plants. Retrieved November 12, 2012 from http://www.ehow.com/about_5474420_effects-caffeine-plants.html Euphrasie, N. (2012). The Effect of Caffeine on Plant Growth . Retrieved November 12, 2012 from http://www.education.com/science-fair/article/effect-caffeine-plant-growth/ Nitrogen Parameters. (2008). About Leaf Color Chart (LCC). Retrieved Green Scale Photo October 17, 2012 from http://nitrogenparameters.com/lcc.html Unknown. (2008). The Effect of Caffeine On Plant Growth. Retrieved September 19, 2012 from http://www.mycaffeineaddiction.com/the-effect-of-caffeine-on-plant-growth