Introduction

Almost all plants are photoautotrophs (Urry et al. 2017, pp. 238-262).
Photoautotrophs refer to organisms that use light as an energy source to
synthesize their own organic molecules from raw materials obtained from the
environment such as CO2, H2O (Urry et al. 2017, pp. 238-262). This process is
called photosynthesis and there are 2 stages involved: Light reactions and the
Calvin cycle (Urry et al. 2017, pp. 238-262). Plant cells contain chloroplasts
where photosynthesis takes place and mitochondria where cellular respiration
occurs (Urry et al. 2017, pp. 238-262). To initiate respiration, sugar and O2 are
required and there are 4 stages involved, releasing CO2 as waste product (Urry
et al. 2017, pp. 238-262). On the other hand, photosynthesis requires light,
water, and CO2. First stage of photosynthesis occurs in the thylakoid
membrane and converts light energy into chemical energy in the forms of ATP
and NADPH to fuel the next stage (Urry et al. 2017, pp. 238-262). In this
process, oxygen is produced as byproduct (Urry et al. 2017, pp. 238-262). This
practical aims to discover how increasing light energy affects the rate of
photosynthesis as measured by rate of oxygen production. When light energy
is present, photosynthesis and respiration occur together. Photosynthesis
raises oxygen concentration while respiration reduces oxygen to water;
therefore, the observed concentration values of oxygen will always be lower
than the actual oxygen concentration produced by photosynthesis. According
to Kong et al., plants grown under high light intensity can maintain higher rate
of photosynthesis by absorbing large number of photons while plants grown
under lower light level tend to have decreased photosynthetic enzymes
activity, which decreases rate of photosynthesis. Under varying light
conditions, plants may enhance light absorption by altering their leaf
structures, photosynthetic pigment contents, or photosynthetic capacity (Kong
et al. 2016, pp. 1-15).
Material and methods
The procedure as set out in the Molecules, Genes and Cells Laboratory Manual
(2021) was followed. An oxygen electrode was provided to measure the
changes in O2 concentration in the chamber containing the spinach leaves. CO2
was provided in the form of NaHCO3. A set of filters and a black cloth were also
provided to decrease light intensity after a steady increasing rate at maximum
light was obtained. However, instead of adding a large pinch of spinach, 2-3
slices of spinach leaves were actually added. Additionally, the rates of oxygen
production were calculated using the software at the end, not in between the
process of changing light intensity.
Results

Figure 1: A scatterplot showing rates of oxygen production in the spinach leaves (Spinacia oleracea) as a
function of increasing light intensity (μmoles photons/m 2s). The blue trend line shows the observed rate of
oxygen production of the spinach leaves when exposed to different light intensities. The orange trend line
shows the photosynthetic rate of oxygen production. The true rates were calculated by subtracting the rate of
respiration (-0.2662μg/L.s) from the observed rates of oxygen production. The scatterplot was drawn using the
obtained data and 2 trend lines were fitted using Excel. These trend lines follow an exponential growth shape.
The light compensation point was indicated on the graph and was found to be approximately 13.5μmoles
photons/m2s.

The scatterplot shows that as light intensity increased, rate of photosynthesis

also increased exponentially.
Discussion
The value of the y-intercept of the blue trend line is -0.2662μg/L.s. This value
represents the rate of respiration or the rate of oxygen consumption in the
dark. In the dark (0 μmoles photons/m2s), only respiration will occur,
consuming oxygen and because there is no further production of oxygen as
byproduct since photosynthesis required light energy to initiate, rate of oxygen
production will appear negative. In the inner membrane of mitochondria,
electron carriers NADH and FADH2 derived from the oxidation of glucose
become the primary electron donors and the donated electrons spontaneously
travel down the electron transport chain (ETC) and reach the terminal electron
acceptor which is oxygen (Urry et al. 2017, pp. 215-237). Oxygen with the extra
electrons then combines with 2 protons in the matrix to form or be reduced to
water (Urry et al. 2017, pp. 215-237).
Light compensation point(LCP) is the light intensity at which photosynthesis
and respiration produce and consume oxygen at the same rate. LCP for spinach
leaves was found to be approximately 13.5μmoles photons/m 2s. The LCP
differs between species and environmental conditions (Wimalasekera 2019,
pp. 65-73). Plants grown in light-limited environment have lower LCP
compared to plants grown in full sunlight (Wimalasekera 2019, pp. 65-73).
Varying light intensities between 0 and 91μmoles photons/m2s resulted in a
sharp increase in the values of both trend lines. Light reactions begin when a
photon of light hits a pigment molecule in a light-harvesting complex of
photosystem II, elevating one of its electrons to a higher energy level (Urry et
al. 2017, pp. 238-262). When this unstable electron falls back to its ground
state, it releases energy and this energy then excites another pigment
molecule (Urry et al. 2017, pp. 238-262). This process continues until the
energy reaches the P680 pair of chlorophyll a and causes one of its electron to
jump to the primary electron acceptor (Urry et al. 2017, pp. 238-262). This
photoexcited electron then spontaneously travels down the ETC (Urry et al.
2017, pp. 238-262). As the pair of chlorophyll now misses one electron, an
enzyme catalyzes the oxidation or the splitting of a water molecule into 2
electrons, 2 protons and 1 oxygen atom. One of the electrons is used to
replace the missing electron. The oxygen atom then combines with another
oxygen atom to form O2 while protons are released into the thylakoid space,
further contributing to the proton gradient. Therefore, increasing light
intensity may result in more production of oxygen.
When light intensity was at 500μmoles photons/m2s, the enzymes involved in
both stages of photosynthesis were all saturated and the rates of those
processing reactions reached maximum; meaning that photosynthesis could
not occur faster, causing rate of oxygen production to plateau compared to the
sharp increase at the start. In addition, according to Shin, Song and Thompson,
oxygen inhibition could be the reason causing the lines to plateau as the
ribulose phosphatase enzyme primarily catalyzing photosynthesis also
catalyzes respiration at higher oxygen concentrations. Although their study on
aquatic plant Elodea canadensis failed to reject the null hypothesis, there was
still an observed trend suggesting that as light intensity increased, rate of
oxygen production also increased and plateaued at a light intensity of 6000 lux
or approximately 111μmoles photons/m2s (Shin, Song & Thompson 2012).
Adding CO2 also increased rate of oxygen production from 2.274 to
4.900μg/L.s. The enzyme Rubisco catalyzes the attachment of each CO2
entering the cycle to a five-carbon sugar called RuBP, resulting in the formation
of 2 3-phosphoglycerate molecules (Urry et al. 2017, pp. 238-262). 6 molecules
of ATP and NADPH produced in stage 1 are used to convert 6 3-
phosphoglycerate molecules into 6 molecules of G3P (Urry et al. 2017, pp. 238-
262). One of the G3P molecules leaves the cycle to contribute to the formation
of glucose while others regenerate RuBP (Urry et al. 2017, pp. 238-262). Both
ADP and NADP+ then return back to the Light reaction to accommodate the
production of ATP and NADPH (Urry et al. 2017, pp. 238-262). Although the
Calvin cycle does not directly require light to occur, the process still requires
ATP and NADPH which are the products of the light dependent reaction. When
there is no light to drives stage 1 of photosynthesis, the Calvin cycle will not
occur since there is no ATP and NADPH produced. Similarly, when there is no
CO2, the Calvin Cycle will not occur; therefore, ATP and NADPH will not be
converted back into ADP and NADP+ required for stage 1 of photosynthesis.
When CO2 concentration increases, the number of successful binding of CO 2 to
Rubisco will increase, causing the rate of the dark reaction to increase and
ultimately, increasing the rate of photosynthesis.
References
Kong et al. 2016, ‘Effects of light intensity on leaf photosynthetic characteristics, chloroplast
structure, and alkaloid content of Mahonia bodinieri (Gagnep.) Laferr.’, Acta Physiologiae
Plantarum, vol. 38, no. 5, pp. 1–15, viewed 16 May 2021,
<https://link.springer.com/content/pdf/10.1007/s11738-016-2147-1.pdf>.

Molecules, Genes and Cells Practical Manual 2021, University of Adelaide.

Shin, D, Song, M & Thompson, C 2012, ‘Turn That Light Up: Examining the Effect of Light
Intensity on Photosynthesis as Measured by Oxygen Production in Elodea canadensis’, The
Expedition, vol. 1, viewed 16 May 2021,
<https://ojs.library.ubc.ca/index.php/expedition/article/view/183642 >.

Urry et al. 2017, ‘Cellular Respiration and Fermentation’ in Campbell Biology: Australian and
New Zealand Edition eBook, pp. 215-237, viewed 16 May 2021,
<https://ebookcentral.proquest.com/lib/adelaide/detail.action?docID=5220583.>.
Urry et al. 2017, ‘Photosynthesis’ in Campbell Biology: Australian and New Zealand Edition
eBook, pp. 238-262, viewed 16 May 2021,
<https://ebookcentral.proquest.com/lib/adelaide/detail.action?docID=5220583.>.
Wimalasekera, R 2019, ‘Effect of Light Intensity on Photosynthesis’ in Photosynthesis,
Productivity and Environmental Stress, pp. 65-73, viewed 16 May 2021,
<https://onlinelibrary.wiley.com/doi/abs/10.1002/9781119501800.ch4#:~:text=Crop
%20production%20is%20strongly%20dependent%20on%20the%20photosynthetic
%20rates.&text=Too%20high%20or%20too%20low,intensity%20and%20reaches%20a
%20maximum.>.