Fertilizer Effects on
Chlamydomonas
Hydrogen Production
BY STEELE MERCER
THE PROBLEM
 Energy sources create lots of pollution
US- 1.7 billion tons of CO2 pollution per year
 The World- 36.33 Billion tons of C02 pollution per year
Can life be utilized as a solution?
Chlamydomonas reinhardtii
Single cell green algae
Can survive in light or dark
Circle or oval-shaped organism with two flagella on
anterior
Found in fresh water, damp soil, the sea and sometimes
even snow
Model organism for scientific tests
How does C. reinhardtii create
Hydrogen?
Deprivation of sulfur switches from Oxygen to
Hydrogen
Activates Hydrogenase enzyme to make
hydrogen
Reaction stopped by Oxygen created in the
sugar process
Oxygen inhibits Hydrogenase
Chemicals
NaNO3
Useful in several
reactions:
Fertilizers
Rocket Propellants
Smoke Bombs
 (NH4)3PO4
 Instability, elusive
and of no
commercial value
(except for
scientific
research)
Purpose
To determine the effects of fertilizer on
Chlamydomonas reinhardtii hydrogen
production.
Hypothesis
Null Hypothesis- Fertilizer exposure will have no significant effect on
Chlamydomonas reinhardtii Hydrogen production
Alternative Hypothesis- Fertilizer exposure will
have a significant effect on Chlamydomonas
reinhardtii Hydrogen Production. Exposure will
increase hydrogen production.
Materials
Chlamydomonas reinhardtii Algae
Sodium Nitrate Solution
Ammonium Phosphate
21 Test Tubes
21 Balloons
10 ml graduated cylinder
Light Rack
1000 ul Pipette
200 ul Pipette
Procedure
1.Solutions
Control
(ml)
1%(NaNo3)
(ml)
.1%(NaNo3)
(ml)
1%
(NH4)3PO4
(ml)
.1%(NH4)3P
O4
(ml)
Algae
2
2
2
2
2
Variable
0V
.1
.01
.1
.01
Spring Water
3
2.9
2.99
2.9
2.99
2. Balloons were affixed on the tube
3. Tubes placed in light rack
4. Light rack was placed in a rack
Procedure
5. Sat in the box with a 12 hour day/night light cycle for 10 days
6. Balloons were detached and inverted under a submerged graduated cylinder
7. The displacement of water was recorded
Start of 10 day period
End of 10 day period
Hydrogen Production(ml)
Ammonium Phosphate [(NH4)3PO4]
0.9
0.8
0.8
0.7
0.6
0.5
0.45
0.4
0.3
0.2
0.2
0.1
0
Control 0%
P Value 0.1%: 0.000423
0.1%
Concentrations
1%
P Value 1%: 0.002452
Sodium Nitrate [NaNO3]
Hydrogen Production (ml)
0.7
0.6125
0.6
0.5
0.45
0.4
0.3
0.15
0.2
0.1
0
Control 0%
P Value: 0.01974
0.1%
Concentrations
1%
P Value: 0.000325
Dunnett’s Test
NaNO3 Concentration
T-value
Significance
[1%]
6.220
Significant
[.1%]
3.37
Significant
T Crit.> 3.29
(NH4)3PO4
Concentration
T Value
Significance
[1%]
4.74
Significant
[.1%]
6.64
Significant
Conclusion
The null hypothesis was rejected for all
concentrations.
All concentrations of chemicals did SIGNIFICANTLY
affect the growth of algae.
However the type of the effect varied
Increased
Decreased
NaNO3 1%
NaNO3 0.1%
(NH4)3PO4 1%
(NH4)3PO4 0.1%
Limitations and Extensions
Limitations
Low number of replicates
and concentrations
Travel time of algae
Gas produced may not
have been exclusively
hydrogen
Extensions
More replicates and
concentrations
Fresher Algae
Test for Hydrogen
Works Cited
"Coal Power Air Pollution." Union of Concerned Scientists. N.p., n.d. Web. 20 Dec. 2015.
"The Element Hydrogen." It's Elemental -. N.p., n.d. Web. 20 Dec. 2015.
"Hydrogen Production by Chlamydomonas Reinhardtii Cells in Sulfur-Deprived Environments. "
OpenWetWare RSS. N.p., n.d. Web. 20 Dec. 2015.
Nenninger, Katie. "Chlamydomonas Reinhardtii." Chlamydomonas Reinhardtii. N.p., n.d. Web. 20 Dec.
2015.
"Surprising Findings on Hydrogen Production in Green Algae." - Uppsala University, Sweden. N.p., n.d.
Web. 20 Dec. 2015.
ANOVA
Anova: Single
Factor
Anova: Single
Factor
SUMMARY
Groups Count
Control
4
High
NaNO3
4
Low
NaNo3
4
Sum
SUMMARY
Groups Count
Control
4
High AP
4
Low AP
4
Average Variance
1.8
0.45 0.003333
0.6
0.15
0.003333
2.45
0.6125
0.007292
Sum
1.8
0.8
3.2
Average Variance
0.45 0.003333
0.2 0.006667
0.8 0.006667
ANOVA
ANOVA
Source
of
Variation
SS
Between 0.44041
Groups
7
Within
0.04187
Groups
5
Total
0.48229
2
df
MS
2
0.220208
9
0.004653
11
F
47.32836
P-value
1.67E-05
F crit
4.256495
Source
of
Variation
SS
Between 0.72666
Groups
7
Within
Groups
0.05
Total
0.77666
7
df
MS
2
0.363333
9
0.005556
11
F
P-value
65.4
4.36E-06
F crit
4.256495