caffeine’s effect on - centralcatholichs.com science/pjas/pjas 15 pdf... · caffeine’s effect...
TRANSCRIPT
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CAFFEINE’S EFFECT ON
MUNG BEAN
GERMINATION AND
GROWTH
TODD ORAVITZ
9TH
GRADE
CENTRAL CATHOLIC
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INSPIRATION
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CAFFEINE
• Naturally occurring substance
• Bitter, white purine compound
• Similar chemical structure to
adenine and guanine
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CAFFEINE EFFECTS
• Blocks adenosine receptors, leading to calcium loss in plant cells
• Low calcium can cause problems with
• Cell membrane permeability
• Cell plate formation
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CAFFEINE EFFECTS
• Interferes with plant cytokinesis
• Stops Golgi vesicles from fusing
with membranes by decreasing
ATP activity
• Has been shown to inhibit cell
division in plants
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CAFFEINE IN NATURE
• Pesticide-like protection to
plants containing it
• Germination of competing
seedlings may be slowed by
plants depositing caffeine in
nearby soil
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GUARANA PLANT
• Effective natural stimulant
• Seeds contain about twice the
caffeine concentration as those
from coffee
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GUARANA PLANT
• Naturally alters one’s
perception of fullness, leading
to weight loss
• FDA recognizes it as “generally
safe”
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PURPOSE
• To determine if caffeine has an effect on germination and growth of mung beans
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HYPOTHESES
• Null
• Caffeine will not have a
significant effect on mung
bean germination and growth
• Alternative
• Caffeine will have a
significant effect on mung
bean germination and growth
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MATERIALS
• Seed starter trays
• Potting soil
• Mung beans
• Guarana –
caffeine source
• Sunlight via
window
• Room lights
• Tap water
• Pyrex 500mL
measuring cup (to
make test solution)
• 10mL measuring
cup (for watering)
• Ruler
• Scientific scale
(no continuous, dedicated light source)
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PROCEDURE
• Planted mung beans
• 72 plants each in test and control
groups
• 5 mL caffeine solution [200mg/L] given
every other day to test group
• 5 mL tap water given every other day to
control group
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PROCEDURE
• Main shoot height of mung beans measured daily for 28 days
• Mung bean mass measured on day 28:
• Plant removed, rinsed with tap water and cut at ground level
• Above and below ground wet masses measured, then added for total
• Procedure repeated after air drying for three hours to obtain dry mass
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CAFFEINE CONTROL
DAY
28
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GERMINATION ANALYSIS
No growth Growth Total
Caffeine 51 21 72
Control 19 53 72
Total 70 74 144
Χ2 = 28.466, p < 0.00001
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CONCLUSIONS
• Null hypothesis rejected
• Alternative hypothesis accepted –
caffeine had a significant effect on
mung bean germination and growth
• Specifically, it significantly decreased
the number of mung beans that
germinated
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QUESTION
• When caffeine group mung
beans did germinate, did they
exhibit similar growth
characteristics to control?
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avg
sh
oo
t h
eig
ht
(mm
)
day
blue-
caffeine
green-
control
CAFFEINE EFFECT ON SHOOT HEIGHT
general
linear
modeling,
p = 0.812
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HEIGHT ANALYSIS
• Daily average mung bean shoot
height compared
• Only plants that germinated
• No significant difference between
caffeine and control average daily
shoot heights
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0
0.1
0.2
0.3
0.4
0.5
0.6
above ground below ground total
caffeine
control
AVG MASS/PLANT – WET
wet mass above ground below ground total
caffeine, g 0.297 0.261 0.558
control, g 0.259 0.308 0.567
p value 0.052 0.035 0.785
significant? no yes no
mass (
g)
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0
0.05
0.1
0.15
0.2
0.25
0.3
0.35
0.4
above ground below ground total
caffeine
control
ma
ss (
g)
AVG MASS/PLANT – DRY
dry mass above ground below ground total
caffeine, g 0.224 0.129 0.353
control, g 0.176 0.142 0.318
p value 0.009 0.262 0.125
significant? yes no no
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ABOVE/BELOW GROUND
WET MASS RATIO
0
0.2
0.4
0.6
0.8
1
1.2
mass ratio
Caffeine 0.297/0.261 = 1.138
Control 0.259/0.308 = 0.841
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MASS ANALYSIS
• T-test done for all 6 subgroups
• Significant difference seen in 2
• Below wet (p=0.035) and above
dry (p=0.009)
• No significant difference in the
other 4
• Above wet, total wet, below dry
and total dry
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HEIGHT, WET MASS CORRELATION
heig
ht
(mm
)
total wet mass (g)
blue – caffeine;
R=0.963
green – control;
R=0.807
p<0.001
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HEIGHT, DRY MASS CORRELATION
heig
ht
(mm
)
total dry mass (g)
blue – caffeine;
R=0.941
green – control;
R=0.815
p<0.001
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HEIGHT VS MASS
ANALYSIS
• Height vs total wet and dry mass
• Only plants that germinated
• Pearson correlation coefficient
• Height correlated with mass in both
wet and dry groups
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CONCLUSIONS
• Null hypothesis rejected
• Alternative hypothesis accepted
– caffeine had a significant
effect on mung bean
germination and growth
• Specifically, it reduced the
number of plants that germinated
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CONCLUSIONS
• Mung beans in the caffeine group
that did germinate, however,
showed similar growth to control
• No significant differences in
• Average daily shoot height
• Average total wet mass
• Average total dry mass
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LIMITATIONS AND
EXTENSIONS
• Limitations
• Did not control soil content
• Short drying time
• Inconsistent lighting
• Extensions
• Different caffeine concentrations
• Correlate pre-planting mung bean mass with germination
• Defined non-sunlight source
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BIBLIOGRAPHY
• ag.arizona.edu/pubs/garden/mg/soils/types.html
• Arnaud, M.J. 1987. The pharmacology of caffeine. Prog.
Drug Res. 31: 273-313.
• Bonsignore, C.L, and Hepler, P.K. “Caffeine Inhibition of
Cytokinesis: Dynamics of Cell Plate Formation-
Deformation in vivo.” Protoplasma. 129, 28-35; 1985.
• en.wikipedia.org/wiki/guarana
• Etherdon, G.M., and M.S. Kochar. 1993. Coffee: Facts and
controversies. Arch. Fam. Med. 2(3):317-322.
• extension.oregonstate.edu/lane/sites/default/files/docume
nts/cffee07.pdf
• Hazardous Substances Data Bank. 1997. Caffeine. HSDB
number 36. Bethesda, MD: National Library of Medicine.
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BIBLIOGRAPHY
• Hepler, P.K. “Calcium: A Central Regulator of Plant Growth and Development.” Plant Cell 2005; 17; 2142-55.
• Kabagambe, Edmond K. "Benefits and Risks of Caffeine and Caffeinated Beverages." UpToDate. Wolters Kluwer Health, 27 Feb 2013.
• Lopez-Saez, J.F. et al. “ATP level and caffeine efficiency on cytokinesis inhibition in plants.” Eur J Cell Biol. 1982 Jun; 27(2): 185-90.
• Nathanson, J.A. “Caffeine and related methylxanthines: possible naturally occurring pesticides.” Science. 226 (4671), 184-7; 1984.
• www.hort.purdue/edu/ext/groundsforgardening.html
• www.hort.purdue.edu/newcrop/afcm/mungbean.html
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ACKNOWLEDGEMENTS
• Thanks to Mr. Krotec for support and guidance throughout the experiment.
• Thanks to James Ibinson, MD, PhD, for help with statistical analysis.
• Thanks to my parents for helping me with ideas and suggestions, as well as supply of materials.
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ANOVA TESTING –
ABOVE/BELOW WET MASS
Anova: Single Factor
SUMMARY
Groups Count Sum Average Variance
Column 1 21 6.233 0.29681 0.004239
Column 2 21 5.489 0.261381 0.006237
Column 3 53 13.718 0.25883 0.008701
Column 4 53 16.345 0.308396 0.007563
ANOVA
Source of Variation SS df MS F P-value F crit
Between Groups 0.078898 3 0.026299 3.588881 0.015321 2.667443
Within Groups 1.055238 144 0.007328
Total 1.134137 147
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ANOVA TESTING –
ABOVE/BELOW DRY MASS
Anova: Single Factor
SUMMARY
Groups Count Sum Average Variance
Column 1 21 4.701 0.223857 0.004027
Column 2 21 2.71 0.129048 0.001461
Column 3 53 9.332 0.176075 0.005073
Column 4 53 7.547 0.142396 0.002345
ANOVA
Source of Variation SS df MS F P-value F crit
Between Groups 0.133358 3 0.044453 12.91884 1.6E-07 2.667443
Within Groups 0.495492 144 0.003441
Total 0.62885 147
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ABOVE/BELOW GROUND
DRY MASS RATIO
0
0.2
0.4
0.6
0.8
1
1.2
1.4
1.6
1.8
2
mass ratio
Caffeine 0.224/0.129 = 1.736
Control 0.176/0.142 = 1.239