Niem Pham

Biology 1B
May 20, 2015
Effects of Caffeine on Raphanus sativus as Determined by Observation of Plant Growth and
Plant Height
Introduction:
Caffeine, a family of chemicals called the xanthines has an effect of increasing the
alertness within humans body along with the neural activity (Watkins, 2011). Because humans
have well developed nervous system, it leaves a great affect on humans, however, plants lack
nervous system, meaning plants do not experience a benefit of alertness or neural activity within
its system. Although caffeine has no effect on plants due to lack of nervous system, it does
however, interfere with the regulation of calcium within cell (Watkins, 2011). A study found that
caffeine also interferes with a cells calcium regulation by inhibiting adenosine receptor.
Adenosine receptors play an important role in controlling calcium levels, and when they are
inhibited by caffeine, cells release large amounts of their internal calcium. When caffeine causes
the cell to release internal calcium, it has the potential to greatly hamper all of these functions of
calcium (Watkins, 2011).
There have also been several other studies that found caffeine may present signs of
inhibition towards plant growth. A study found that caffeine could have a role in competition, by
inhibiting the germination of other seeds (Suzuki and Waller, 1987). This supports the idea that
caffeine may be a growth inhibitor for non-caffeine-producing plants. Caffeine also has an ability
to induce the chromosomal aberration which may affect seeds during the start of germination
(MYK Ansari, 2009).

A different study suggests that caffeine (C8H10N4O2) is a chemical compound that is

catabolized into ammonia (NH3), a source of nitrogen and carbon dioxide. The study also found
that younger leaves had a higher caffeine content than the older leaves, which suggests that the
older leaves catabolized the caffeine (Mazzafera 2004). This supports the fact that caffeine may
be involved in plant growth as a potential source of nitrogen and carbon dioxide.
With the research provided we were able to produce multiple hypothesis for the different
possible outcomes. A plausible prediction is caffeine will support plant growth by providing a
source of nitrogen or carbon dioxide through its the catabolic breakdown (Hypothesis 1).
Another possible hypothesis one can draw from the research is that the caffeine will inhibit or
decrease growth (Hypothesis 2). These hypothesis are both assuming that Raphanus sativus will
be able to catabolize the caffeine the same way caffeine producing plants can. If the radish plant
cannot catabolize the caffeine, then it may have no effect on growth (Hypothesis 3).
Methods:
To test our hypothesis we set up an experiment where we observed the effect that caffeine
has on plant growth. In the experiment, the dependent variables was our measurements and
observations of the health and growth of the plants. The independent variable was the different
concentrations of caffeine given to the plants. The amount of water, light exposure, soil type,
plant type and caffeine type were the most important control variables. There were five different
groups with ten radish seeds each, with 3 pots per group. We gave four of the groups different
concentrations of caffeine while the control group only received tap water. First, we planted the
radish seeds and let them germinate without any external influences. Next, we made our solution
by mixing the specified concentration caffeine (powder) with 50mL of tap water in a plastic
container. After germination, we poured 5mL of different concentrations of caffeine solution

(0.4g/ 50mL, 0.3g/50mL, 0.2g/50mL, and 0.1g/50mL) using a graduated cylinder to each pot in
the experimental groups. Solution was added every lab section.
We measured the plants height by measuring from the top of the soil up to the apical
bud. Pictures of each group were taken at set intervals of time to show effect of caffeine on
plants visually.
Results:
Our data was recorded both in quantitative and qualitative observations. We measured the
plants everyday (possible) that we were in lab, which ended up being six days. The quantitative
observations was height measured in centimeters and is portrayed in Figure 1. Figure 1 is the
average of the plant height. Statistical information is also included (Figure 2) in the form of a
simple t-test. The qualitative observations will be described. Day 1: All the seeds had
successfully germinated and looked rather healthy. Day 2: All the seedlings still appear healthy
and nearly identical as Day 1. Day 3: All the seedlings are still healthy and the average height of
all plants about doubled. Day 4: There is a much higher variance in plant height. Overall plant
health seems just slightly degenerated: slight yellowing of leaves and stems seems skinnier. A
couple plants (of the same pot) from the control group are wilted. Soil was very dry because we
did not add enough water to last the entire weekend. Day 5: Overall plant health are similar to
Day 4.The same plants that appear wilted on Day 4 are still wilted including 1 more plant in the
400 mg concentration group. Day 6: The same plants that appeared wilted on Day 5 are still
wilted including 1 more plant in the 300 mg concentration group. The overall health of the plants
that received treatment seem slightly more deteriorated: stems appear a little skinner. The
petioles of the higher concentrations seem shorter.

Figure 1

type

0.4g/50mL

0.3g/50mL

0.2g/50mL

0.1g/50mL

t calculated

0.401

-0.740

-1.251

-1.356

t critical

1.8124

1.8124

1.8124

1.8124

Figure 2: T test values for each concentration growth rate.

Discussion:
In the experiment we noticed that the plants receiving the caffeine treatments tended to
have skinnier less healthy appearing stems. They also appeared to have shorter petioles and slight
yellowing of the leaves, especially the higher concentrations. But oddly the highest concentration
treatment (400 mg) had the highest plant height, even taller than the control plants. All of the
other treatments grew at around the same height and had very minimal health detriment. These
results appeared random and seemed to have no type of correlation, other than the fact that they
were all shorter than the control plants (Figure 1). Our data showed no correlation between
growth pattern and the concentration of caffeine which can be observed in Figure 2. The t-test

values shows no significant effect on growth rate of different concentration of caffeine because tcalculated values are smaller than t-critical for all concentrations.
According to Campbell, leaves of a plant that do not have enough calcium and nitrogen
will crinkle and turn yellow on older leaves. The older leaves that were turning yellow reminded
us of the sunflower experiment. But the yellowing in older leaves was results of nitrogen
deficiency, not calcium which was interesting to observe because we believed that it might show
the effects of calcium deficiency where the newer leaves may turn yellow. This information
discredits hypothesis (1) because the plants showed signs of nitrogen deficiency, meaning
caffeine did not provide a source of nitrogen. Or this could also provided a bases for hypothesis
(3) because Raphanus sativus may not have been able to catabolize the caffeine into ammonia
(further broken down into nitrogen). On the other hand this supports hypothesis (2) because the
caffeine caused detriment towards the plant health, but the plant experienced nitrogen deficiency
rather than calcium deficiency.
While performing our procedures we experienced a couple experimental errors that were
very important. One problem was the fact that the caffeine powder did not dissolve in water, so
we ended up using a mixture rather than a solution. This may have possibly ruined the entire
experiment because the caffeine was still in powder form when it was in the soil, so we do not
know if it was actually absorbed by the plant. The second major error was our inability to
maintain water as a control variable. We watered the plants unevenly and also did not supply all
of the plants with enough water to last the entire weekend. This could account for the plants that
experienced diminished health and could also explain why the 400 mg group had the highest
recorded height. This was expressed in Day 4 of the qualitative data (control group).

Because of our experimental errors, the results are inconclusive and do not provide
absolute proof towards any of the hypothesis. The following is a deduction. If the health
problems we observed were from our watering error than hypothesis (3) may be correct. This is
because the plant did not absorb or was not able to break down the caffeine, thus the caffeine had
no effect. But if the health problems were a result of the caffeine rather than dehydration then
both the hypothesis (1 and 3) are incorrect because the plant did in fact absorb the caffeine and it
caused detriment rather than growth. If this is true, then hypothesis (2) is correct.
Because there is no discernable pattern in plant growth with respect to caffeine
concentration along with our experimental errors, we concluded that there is no distinguishable
correlation between caffeine and plant growth.
If we were given the chance to redo the experiment, we would like to water our plants
evenly and regularly enough so the plants would not drying out, which may have affected our
results. We would also completely dissolve the caffeine so we know for sure that the caffeine was
absorbed. Another thing we would like to attempt is having a caffeine-producing plant as one of
the controls in order to see how additional caffeine affects a plant the we readily know can
catabolize caffeine.

Literature Cited

Campbell, N.A., J. B. Reece, L. A. Urry, M. L. Cain, S. A. Wasserman, P. V. Minorsky,

and R. B. Jackson. 2008. Soil and Plant Nutrition. In Campbell et al., 804-805.
Benjamin Cumming, San Francisco, California, USA.
This chapter, Soil and Plant Nutrition, in textbook provides how calcium and iron affect plants
structure and growth. This supports the experiment result and shows low level of calcium and
iron influenced the result.
Hollingsworth, R.G., J.W. Armstrong and E. Campbell. 2003.Pest control: caffeine as a
repellent for slugs and snails. Nature 417: 915-916.
This source provides information that supports hypothesis (1) by detailing how caffeine in high
concentration may serve as a natural pesticide against insects.
Mazzafera, Paulo. 2004. Catabolism of caffeine in plants and microorganisms. Frontiers in
Bioscience 9: 1348-1359.
This source explains how caffeine-producing plants catabolize and utilize caffeine as a potential
source of carbon dioxide and nitrogen. This source also supports hypothesis (1).
Suzuki, T. and G.R. Waller. 1987. Alleopathy due to purine alkaloids in tea seeds during
germination. Plant Soil 98: 131-1.
This source explains how caffeine released into the soil by tea plant seeds may inhibit the growth
and germination of other seeds. This source supports hypothesis (2).
Watkins, K., 2011. Caffeines Effect on Plants Germination. Journys. para. 1-2

This article is about effect of caffeine plant germination by decreasing calcium level which is
relevant to my project. This shows some ideas of how caffeine works on human and plants.