Abstract:

Photosynthesis is a process used by plants and other organisms to convert light energy,

water and carbon dioxide into energy for the organism. These reactants are essential for the

overall reaction; the hydrogen from the water oxidizes the nicotinamide adenine dinucleotide

phosphate, the carbon from the carbon dioxide forms carbon chains in the stroma during the

Calvin Cycle, and the light energy from the sun initiates and propels the entire reaction.

However, with the increase of carbon dioxide on Earth, due to the rise of population, oil

refineries and the overall pollution of green house gases, it becomes increasingly necessary to

gauge the effect of the carbon dioxide on the process of photosynthesis in-depth. In this

experiment, the concentration of the solution sodium bicarbonate is varied to observe the results

of an increased amount of carbon in the reactant component of photosynthesis. First, Spinacia

oleracea (spinach leaves) were obtained. Five conditions were organized, each having different

grams of sodium bicarbonate in the same amount of tap water (100 mL), hence creating variation

among the concentrations; the first beaker, which was the control, contained 0.75g of NaHCO3,

the second contained 1.25g, the third contained 1.75g, in the fourth there was 2.25g, and in the

fifth, there was 2.75g. Ten disks were cut out from the spinach leaves to place in each beaker,

and then these beakers were placed 25cm from a light source. The results reflected the

hypothesis; as the concentration of carbon increased in the solution, the amount of floating leaf

disks increased.

Introduction:
All the oxygen that is utilized by all living creatures today originated from

photoautotrophs; the air that dolphins breathe when they jump across the horizon, that babies

puff once they enter this world for the first time, and that dogs huff after long walks, all comes

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from organisms that carry out photosynthesis (1). It is no debate that the process of

photosynthesis has been a major part of the survival of living organisms on the planet Earth and

will continue to facilitate the existence of life in the future. However, in recent decades, the level

of carbon emissions of the planet has increased steadily (2). Therefore, the photoautotrophs,

which life on Earth has depended on for centuries at the least, may be affected, because carbon is

a main part of photosynthesis (3).

That being said, the majority of research being done with photoautotrophs is more

concerned with the effects of temperature and sunlight rather than the effects of carbon. Jing and

his team researched the physiological, morphological and yield-related responses to night

temperature changes in plants across different species and types. They determined that

high-night temperature and low-night temperature facilitated changes in the physiology and

morphology of plants over time. Soleh and his team took a similar approach. Interested in how

the speed of the induction response of photosynthesis in high light affects the cumulative carbon

gain if the plant as well as its impact on the growth and yield. However, they only tracked the

chemical reactions that were motivated by an increase in light, thus failing to categorize the

direct effects of carbon on photosynthesis. Chen and his team observed that the photosynthetic

pathway of plants might play an important role in regulating soil respiration. Thus, particular

wetlands could fix significant amounts of carbon in the soil.

It seems that although carbon has a vital role in photosynthesis, its direct effects in the

reaction has been neglected from recent research proposals. Perhaps this is due to the fact that

carbon does not have the morphological or physiological impact on plants or the yield in general.

Therefore, it is less imperative to delve into the direct role that carbon has on photoautotrophs.

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However, one could argue with the global epidemic that is global warming, the rise of carbon

emissions on an international scale is clear, which means that carbon has a greater impact on

photosynthesis, thereby the process by which we all breathe and, indirectly, consume energy. In

this way, this experiment is of upmost relevance because of its geopolitical and environmental

implications for the health and wellbeing of humanity.

Methodology:
In order to observe the effects of carbon on the process of photosynthesis, five beakers,

each containing varying concentrations of sodium bicarbonate, were set up and observed under a

light source. Fifty disks were punched out from spinach leaves, and an equal number was placed

into each beaker. In order to remove the oxygen from the leaf disks, a syringe plunger was used

to create a vacuum, allowing the air to flow out of the spaces in the lead disks (Figure 1).

Figure 1: Syringe plunger

Leaves float at the top of the syringe before removing the oxygen from the disks. Pulling the
plunger down, a finger is placed at the mouth of the syringe, creating a pressure. Then the mouth
is released, expelling all the oxygen from the syringe and the leaf disks. The solution is then
placed in its respective beaker.

The first beaker was the control. It contained 0.75g of sodium bicarbonate in 100mL of

tap water, so the concentration of sodium bicarbonate in this beaker was 0.75%. The second

beaker contained 1.25g of sodium bicarbonate in 100mL of tap water, or a concentration of

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1.25%. The third beaker contained 1.75g of sodium bicarbonate in 100mL of tap water, which

means it had a concentration of 1.75%. In the fourth beaker, there was 2.25g of sodium

bicarbonate along with 100mL of tap water. The concentration of sodium bicarbonate in this

beaker was 2.25%. In the fifth and final beaker, there was 2.75g of sodium bicarbonate coupled

with 100mL of tap water, which yielded a concentration of sodium bicarbonate that was 2.75%.

The beakers were observed every minute for eighteen minutes (Figure 2).

Figure 2: Experimental Setup

The experimental setup of 2.75g/mL and 2.25g/mL beakers alongside the experimental setup of
1.75g/mL and 1.25g/mL beakers. The control beaker (0.75g/mL) was done on a different day,
and did not share a light source with another beaker. The light source was 25cm from the
beakers.

Results:
In the control beaker (0.75g/mL), leaf disks failed to even begin to float until the tenth

minute; by the eighteenth minute, five leaf disks had floated to the surface of the solution. In the

beaker with 1.25% concentration, the first leaf disk floated at the fifth minute. By the end of the

eighteenth minute, almost all of the leaf disks were floating at the surface. In beaker that had a

sodium bicarbonate concentration of 1.75%, a leaf disk was already floating upon entering the

solution. By minute eighteen, all but one leaf disk were floating at the top of the solution. In the

beaker with a concentration of 2.25%, the first leaf disk was started to float in the eighth minute,

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which was later than the beakers with lower concentrations. By the end of the trial, eight leaves

were floating at the top of the solution. In the beaker with a concentration of 2.75%, eight leaves

were floating in the solution even before the trial was started. By the end, all the leaves were

floating at the surface (Figure 3 and Figure 4).

[NaHCO3] vs. Amount of Disks that Rise to the Surface over time
Time min 1 2 3 4 5 6 7 8 9 10 1 1 1 1 15 16 17 18
1 2 3 4
[NaHCO3
]
0.75g/mL 0 0 0 0 0 0 0 0 0 0. 3 3 3 5 5 5 5 5
5
1.25g/mL 0 0 0 0 1 1 3 3 3 3 6 6 6 7 9. 9. 9. 9.
5 5 5 5
1.75g/mL 1 1 1 1 1 1 4. 5 5 5 5 5 5 5 7 9 9 9
5
2.25g/mL 0 0 0 0 0 0 0 0. 1 1 1 2 5 6 6 7 7 8
5
2.75g/mL 8 8 9 9 9 9 10 10 1 10 1 1 1 1 10 10 10 10
0 0 0 0 0
Figure 3: Table of Values
Depiction of how the varying concentrations of sodium bicarbonate affect the number of leaves
that rise to the surface of the solution over time. Half values refer to the leaves that were nestled
in between the top of the solution and the bottom.

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Figure 4: [NaHCO3] affects the number of floating disks in the solution over time
Line graph analysis of the number of leaves that float in each beaker and at which times.

In the first beaker, which had a sodium bicarbonate concentration of 0.75%, the first leaf

started floating at the tenth minute (Figure 5). By the eighteenth minute, five leaves were floating

at the surface of the solution. The slope of the line, which represents the amount of leaves that

floated in relation to time, or the rate of the reaction, was ~0.38 disks per minute. So, in a

solution with a sodium bicarbonate concentration of 0.75%, approximately 0.38 leaf disks

floated every minute.

Figure 5: 0.75% [NaHCO3] affects the number of floating disks in the solution over time

In the beaker with a sodium bicarbonate concentration of 1.25% the first leaf disk floated

at the fifth minute. By the eighteenth minute, all leaf disks in the solution were floating; one was

submerged in the middle of the solution. The rate of the disk floatation was larger than the first

beaker, approximately 0.67 leaf disks per minute.

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Figure 6: 1.25% [NaHCO3] affects the number of floating disks in the solution over time

The third beaker, which had a sodium bicarbonate concentration of 1.75%, had a leaf disk

floating at the surface before the trial was even started. By the end, all but one of the leaves had

reached the top of the solution. However, for every minute, approximately 0.48 leaf disks

floated, a rate that is lower than the rate of leaves in the second beaker, which had a lower

concentration of sodium bicarbonate.

Figure 7: 1.75% [NaHCO3] affects the number of floating disks in the solution over time

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 In the fourth beaker, which had a sodium bicarbonate concentration of 2.25%, the first

leaf started to float at the eighth minute. By the end of the trial, eight leaves were floating at the

surface of the solution. The rate of floatation of the fourth beaker was approximately 0.51 disks

per minute; though it is higher than the rate of flotation of the third beaker, it is still significantly

less than the rate of floatation of the second beaker.

Figure 8: 2.25% [NaHCO3] affects the number of floating disks in the solution over time

The fifth and final beaker, which had a sodium bicarbonate concentration of 2.75%, had

eight leaf disks floating at the top of the solution before the trial begun. At the eighteenth minute,

all leaves had reached the top of the solution. The rate of floatation of this beaker was the lowest

of all beakers, approximately 0.11 disks per minute.

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Figure 9: 2.75 [NaHCO3] affects the number of floating disks in the solution over time

Discussion:
The purpose of our experiment was to investigate the effects of the concentration of

sodium bicarbonate on the process of photosynthesis by observing the number of spinach leaf

disks that were floating in the solution over time. It was hypothesized that the solutions with

higher concentrations of sodium bicarbonate would also have the largest number of leaf disks

floating in the shortest amount of time. The data trends suggest that as the concentration of

sodium bicarbonate has a positive relationship with the number of spinach leaf disks that float in

the solution; the beakers with higher concentrations had a greater number of disks floating

sooner than beakers with lower concentrations.

Although the major trend of the data suggests that an increase in the concentration of

sodium bicarbonate directly leads to an increase in the number of leaf disks that float in the

solution, there is a minor exception worth noting. The second beaker, which has a sodium

bicarbonate concentration of 1.25%, had more leaves float faster than the fourth beaker, which

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had a significantly higher sodium bicarbonate concentration. It also had a higher rate of flotation

than the third beaker. Perhaps, this suggests that there is an ideal level of sodium bicarbonate

concentration that yields the highest rate of photosynthesis. That being said, this circumstance

does not break the trend because the second beaker still has a higher rate of floatation than the

control, so it still coincides with the conclusion that higher concentrations of sodium bicarbonate

yield higher floatation rates.

The data also suggests that the concentration of sodium bicarbonate had an impact on

how fast the leaf disks began to float in the solution, or the speed of the photosynthesis reaction.

Although, the final beaker, which had the highest concentration of sodium bicarbonate of all the

beakers, had the lowest rate of floatation, but the initial number of leaf disks was eight. The same

trend is replicated in the third beaker, but it only had one leaf disk floating prior to the start of the

trial. Furthermore, all experimental conditions had their first leaf disks floating sooner than they

started floating in the control. Perhaps, the increase of sodium bicarbonate also increases the

speed of the photosynthesis reaction, as well as the rate and the magnitude.

Ellie Story and Madeline Lloyd were also concerned with how the concentration of

carbon affected the rate of the photosynthesis, represented by the number of leaves that floated in

the solution. Similar to this experiment, they cut out disks from spinach leaves, and placed an

even amount into beakers with varying levels of sodium bicarbonate concentration. The two

experiments differed here: the concentration of sodium bicarbonate in their first beaker was

0.25%, the second had a sodium bicarbonate concentration of 0.75%, the third had a

concentration of 1.25%, and their final beaker had a concentration of 1.75%. The trend that was

observed in this experiment was also noticed in the experiment of Story and Lloyd. They noted

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that the beakers that had higher concentrations of sodium bicarbonate had an increased number

of leaf disks float in the solution. They also observed that the beaker with a 0.25% concentration,

even lower than the control, had less leaf disks float throughout the trial. Therefore, they

concluded that an increased concentration of sodium bicarbonate increases the rate of

photosynthesis.

Some limitations prevented the experiment from precisely replicating a photosynthesis

reaction, hence blurring the some aspects of the relationship between the concentration of

sodium bicarbonate and number of floating lead disks. First, the room in which the experiment

took place was too bright at the time. Therefore, the source of light was uncontrolled and the

results were most likely skewed, especially because this is a photosynthesis reaction, and light

energy propels the reaction. The spherical shape of the light source also limited the experiment to

an extent. Because the light was emitted at an angle, it was not directly focused on the beaker, so

the results of the photosynthesis reaction may have been skewed to some degree.

Some errors in the design stifled the potential of the experiment and probably skewed the

results. The method of removing the oxygen from the leaf disks was difficult and problematic. It

is difficult to be certain of how many pumps of the syringe are needed to fully remove the

oxygen from the leaves. Also, an excess of pumps could damage the results of the experiment, so

the syringe mechanism promoted several logistical issues and altered the results. In addition to

the syringe, the cutting of the spinach leaves into circular disks may have also caused some error.

Some stomata, holes under leaves, may have been cut off, leading to inefficiencies in the leafs

ability to respire. The palisade layer, which was a square shape, could have also been split due to

the circle cut that was recommended, and because, the palisade layer contains the chloroplasts,

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this could have directly interfered with the photosynthesis reaction. The xylem and the phloem

could have also been victimized by the circular cut of the leaves. Although the veins that contain

the xylem and the phloem, the structures responsible for carrying water and food throughout the

leaf respectively, has a cylindrical shape, the circular cut could have still cut off the continuity of

the structures, unnecessarily damaging the leaf and skewing the results.

The experiment could benefit from several improvements. The first would be the number

of the trials. Although certain trends could be observed, more trends would create a greater sense

of certainty in the data as well as an increased statistical relevance. More trials would also

increase the certainty of the data. Though unlikely, there is a possibility that our results are due

to chance because only one trial was done for each experimental condition. A greater number of

trials would denounce that speculation. The experiment also could have benefitted from being in

a darker room. The multiple light sources in the room where the experiment was conducted

probably interfered with the rate of photosynthesis because light energy plays such a huge role in

the reaction. A dimmer laboratory setting would allow the focus to be solely on an individual,

controlled light source, which would lead to less biased results.

Nonetheless, the experiment was successful. The reaction of photosynthesis was

adequately replicated and the effects of the concentration of sodium bicarbonate on the number

of leaf disks that floated in the solution was observed and analyzed. Ultimately, the results

suggest that an increase of the concentration of sodium bicarbonate yields an increase in the

speed, rate and magnitude of photosynthesis. Our results do make rational sense. Carbon is one

of the reactants in the photosynthesis reaction, so it makes sense that when the concentration of

carbon is increased, the speed, rate and magnitude of the reaction increases as well. This is good

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news for a planet that continues to increase its carbon emissions year after year. Perhaps trees,

our sole source of oxygen, will be positively affected by climate change. That being said, it is

imperative that more research on the effects that increased concentrations of carbon have on the

process of photosynthesis, so that the world can be sure that its environmental negligence will

not jeopardize its ability to safely inhale.

Acknowledgements:
Many thanks to all those who assisted in the experimental design, experimental setup,

data acquisition and data organization and presentation: Devyn Prieto, Saira Chowdhry and the C

Band AT Biology class.

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4. Jing et al. Plant Physiological, Morphological and Yield-Related Responses to Night

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plant science (2016).

5. Soleh et al. Identification of large variation in the photosynthetic induction response among 37
soybean [Glycine max (L.) Merr.] genotypes that is not correlated with steady-state
photosynthetic capacity. Photosynthesis Research (2016).

6. Chen et al. Does the different photosynthetic pathway of plants affect soil respiration in a
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