Photosynthesis: Effects of Light experiments, the various samples yielded

different intensities in response to their exposure

on Chlorophyll concentration, to light. The starch test resulted in green discs
Photosynthesis, and Starch testing positive with the presence of starch. The
discs that were exposed in bright light yielded a
Synthesis
more distinguishable positive result compared to
Miguel Angelo Habaña1 | Francheska the ones that were placed in the shaded area. In
Luise Hernandez2 | Andrea Beatrice conclusion, photosynthesis is a monotonous
Limjoco3 | Cristel Marie Madayag4 | Ma.
process that requires some substances in
Isabelita Oran Madridano5 | Anarissa
Mariel Pescasio6 different settings.
Department of Biological Sciences, College of
Science, University of Santo Tomas, España, Manila,
1015 Philippines

1 | Introduction
Abstract
Photosynthesis is an essential process,
Photosynthesis focuses on the particularly for plants because it allows them to
production of specific substances that are absorb light, carbon dioxide, and water to
necessary for sustaining the plant and other synthesize their own food. With the carbon
contributors. The photosynthetic process was dioxide and water present in the air and soil, the
utilized mostly through the photon-harvesting plant intakes the said molecules. For intake, the
mechanism of the chlorophyll. The carbon dioxide and water in the air or soil are
measurement of chlorophyll concentration will reduced and oxidized, respectively. The water
be done through a spectrophotometer. In the molecules are converted into oxygen, while the
experiment, phenol red is utilized as a pH carbon dioxide is converted into glucose.
indicator to determine photosynthesis's Glucose is responsible for providing the energy
regulating effect. The difference in the intensity needed in photosynthesis. These processes
of color between plant tissue samples enables occur within chloroplasts where molecules are
us to determine which of the samples had the absorbed and converted. In photosynthesis, the
highest photosynthetic action during their light light energy absorbed is converted to chemical
exposure. In addition, photosynthesis enables energy through carbohydrates which makes use
the production of starch making it a product of of water and carbon dioxide. Oxygen becomes a
photosynthesis. Nonetheless, variegated plants by-product of this process.
possess non-photosynthetic parts that do not
The chloroplast is where photosynthesis
have similar functions compared to green plants.
occurs in plant cells, but the interactions of light
In relation to this, starch content between green
and pigments take place in the thylakoids.
and white portions of the leaf is different and this
These are membranous discs that are
will be determined utilizing a starch test. Based
connected to each other and contain thylakoid
on the results of varying wavelength
membranes that have chlorophylls and
photosynthetic pigments. The function of Optical Density (OD), also known as
pigments is to absorb light with specific absorbance, is the transmission of light by
wavelengths while deflecting other wavelengths. matter. This is a crucial factor because it can
Depending on the pigments that different plants depict the growth and metabolism of cells. For
contain, the leaf color may vary. In this this experiment, the wavelengths, 663nm and
experiment, three (3) types of photosynthetic 645nm will be used. The formulas to be used will
pigments will be studied and tested, Chlorophyll be presented in the methodology section of the
A, Chlorophyll B, Carotenes, and Xanthophyll. If report.
a plant contains chlorophyll A, it is expected that
Photosynthesis in aquatic plants absorb
the colors produced would be bright green to
the carbon dioxide present in the air or water.
blue-green. For chlorophyll B, yellow green to
The interaction of carbon dioxide and water
olive green is expected. Carotenes should give
produces carbonic acid. In turn, hydrogen ions
out a dark yellow or yellow-orange color and
(H+) are released which decreases the solution
lastly, xanthophyll will just give out a light-yellow
pH level (Hale, 2018). In this experiment, the
color.
photosynthetic conduction of a submerged plant
Light is an extremely important factor in like Hydrilla will be observed. In line with this,
the environment that can cause the manipulation the photosynthesis rate of plants is analyzed
of a plant’s morphological and physiological through utilizing pH indicators like bromothymol
processes. These processes include blue and phenol red. Phenol red or
photosynthesis and chlorophyll synthesis. There phenolsulfonphthalein gradually changes color
is a relationship between light and chlorophyll depending on the pH level (National Library of
concentration because chlorophyll is responsible Medicine, 2022). Under acidic (pH < 7)
for absorbing the light that meets the conditions, the solution turns from yellow to red.
chloroplasts present on the leaves. Depending Under basic (ph > 7) conditions, the solution
on the amount of light received by the plant, the turns red or pink.
concentration of chlorophyll may vary (Johnson
Carbon fixation and reduction in algae,
et al., 2005).
cyanobacteria, and algae occurs due to the
A spectrophotometer is an instrument Calvin-Benson (CB) cycle or C3 cycle.
used to measure the light absorption of a (McFarlane et al., 2019). The cycle happens in
sample. During this process, a beam of light is the inner space of chloroplasts called stroma
emitted, and it goes through the solutions where and produces three-carbon sugars like 3-
it is absorbed depending on the wavelength set. phosphoglyceraldehyde (PGAL) that act as a
A spectrophotometer contains a spectrometer structural building material for organic molecules
that produces a beam of light with the set (e.g., starch, glucose, sucrose, etc.) Starch
wavelength and a photometer that measures the synthesis, also occurring in the stroma, exists as
amount of light that goes through the sample. amylose and amylopectin. To test for its
presence, a starch test is performed through the
addition of iodine-potassium iodide (I2KI)
solution.

Variegation in plants refers to the

display of a variety of colors in their leaf tissue;
most commonly as white (containing no
chlorophyll) and green (containing chlorophyll). It
is caused by a mutation that causes both
defective and normal plastids to be exhibited in
the same but different sector of the plant tissue
(Li et al., 2019). In this report, the researchers
FIGURE 1. Spathiphyllum wallisii setups for high
aim to: 1) determine the concentration of
light intensity (A) and low light intensity (B)
chlorophyll in a plant tissue, 2) discuss the light
intensity effect on chlorophyll concentration of For the experiment proper, 0.25 grams
plants placed in low-light and high-light of sample from fully expanded young leaves of
environments, 3) observe the plant S. wallisii were obtained with the weight
photosynthetic rate at different wavelengths, and measurements done using an analytical
4) assess light and chlorophyll function in starch balance. Following that, leaf sample was cut into
synthesis. smaller pieces and macerated in 10 mL of 80%
acetone using a mortar and pestle. The resulting
mixture was then filtered into a 25-mL volumetric
2 | Materials and Methodology flask. The process of extraction was repeated

Part 1: Effects of light intensity on leaf until the leaf sample appears devoid of green

chlorophyll concentrations color. For each plant, 3 replicates of the leaf

extract were made summing up to a total of 6
Two potted plants of Spathiphyllum replicates for this part of the experiment.
wallisii were each subjected in two different light
setups, with one exposed to sun and the other Absorbance (Optical density) of each

under shade, for a span of 4 weeks. Both the replicate, at 663nm and 645nm, were measured

plants were watered up to their saturation point using a spectrophotometer with 80% acetone

during the entire course of the initial setup. set as the solvent blank for this run. Absorbance
readings obtained from this were used to
calculate the amount of chlorophylls a and b,
total chlorophyll content, and chlorophyll a/b
ratio of each replicate of S. wallisii. Formulae
used to compute for these chlorophyll
concentrations (mg/g) were:
 Chlorophyll a (mg/g)
V
¿ 12.7 ( A663 )−2.69 ( A 645 ) × Part 2: Measuring photosynthesis under
1000 ×W
varying wavelengths of light
(1)
Hydrilla stems were used to observe
Chlorophyll b (mg/g)
photosynthesis under varying wavelengths of
V
¿ 22.9 ( A645 )−4.68 ( A 663 ) × light. In large test tubes, 30 ml of distilled water
1000 ×W
were added in which one test tube will serve as
(2)
control. Six different setups were prepared prior
Total Chlorophyll (mg/g) to the experiment proper and are as follows:

V A. Control (No baking soda, no

¿ 20.2 ( A 645 ) +8.02 ( A 663 ) ×
1000 ×W wrapped)
(3)
B. Baking soda, wrap in foil*

C. Baking soda, not wrapped*

Chlorophyll a/b ratio
D. Baking soda, wrap with red

¿
Chlorophyll a ( )
mg
g
cellophane*

Chlorophyll b (
g )
mg E. Baking soda, wrap with blue
cellophane*
(4)
F. Baking soda, wrap with green
cellophane*

*For setups B - F, 1 gram of baking

soda were added to each tube.

Afterwards, three to four drops of 0.1%

phenol red solution were added to each test
tube before submerging the 2 inches long
Hydrilla stem. Each test tube was then capped
with cork then sealed with parafilm. It is to note
that setups which required foil or cellophane
were wrapped with only one layer of each.
Beakers were also used to maintain an upright
position for the test tubes. All setups were then
FIGURE 2. 25 mL Leaf extract of the 3 replicates
exposed to light (10W LED) for 30 minutes and
from the leaves exposed to high light intensity coloration of the solutions were observed. At
specific times, the color of the solution and its
intensity in each test tube were recorded. For D. 20 mL distilled water, 4 white discs, in
easier observation of coloration and intensity, dark
the test tubes were placed in front of a white E. 20 mL 5% glucose solution, 4 green
background. discs, in bright light
F. 20 mL 5% glucose solution, 4 green
discs, in dark
Part 3: Role of chlorophyll and light in starch G. 20 mL 5% glucose solution, 4 white
synthesis
discs, in bright light
H. 20 mL 5% glucose solution, 4 white
discs, in dark

The discs were floated upside down in

all treatments for 48 hours. The leaf discs were
then prepared in separate labeled test tubes for
the starch test.

30 mL of distilled water were added in

all 8 labeled test tubes which contained the
treated leaf discs. The test tubes were then put
into a water bath of boiling water for 5 minutes to
heat. Water was then poured out then 30 mL of
FIGURE 3. Dieffenbachia seguine after being
places in the dark for 48 hours 70% ethanol was subsequently added into the
same test tubes still containing the treated and
Dieffenbachia seguine, a variegated
heated leaf discs. The test tubes were heated
plant, was used to study the role of chlorophyll
again in a hot water bath until the leaf discs
and light in starch synthesis. D. seguine was
were bleached. Afterward, the bleached leaf
placed in the dark for 48 hours prior to the
discs were rinsed with distilled water and put
experiment proper. Afterward, leaves were
into separate labeled Petri dishes. The leaf discs
detached from the plant. 16 discs were then cut
were then flooded with iodine solution for 1
from the green and white part of the variegated
minute. Excess iodine solution on the leaf discs
leaves using a straw. The 16 green discs and 16
was removed through rinsing with distilled water.
white discs were then distributed to 8 treatments
Changes in color, its intensity, and distribution of
and are as follows:
coloration on the leaf were then recorded for
A. 20 mL distilled water, 4 green discs, in analysis.
bright light
B. 20 mL distilled water, 4 green discs, in
dark 3 | Results
C. 20 mL distilled water, 4 white discs, in
bright light
Part 1. Effects of light intensity on leaf chlorophyll a/b ratios of these shaded plants
chlorophyll concentrations were lower than those of plants exposed to high
light intensity.
The effect of light intensity on the leaf
chlorophyll concentration of Spathiphyllum
wallisii was determined by measuring the
absorbance of the leaf extracts, from the leaves
exposed to high and low light intensity, at 663
nm and 645 nm. From there the concentration of
chlorophyll a, b, total chlorophyll content, and
chlorophyll a/b ratio was calculated (Table 1).

Table 1. Leaf Chlorophyll concentrations plants

exposed to high and low light intensities.

Replica Absorba Absorba Chloroph Chlor Total

te nce nce at yll a ophyll Chlorophyll
(663nm) 645 nm (mg/g) b (mg/g)
FIGURE 4. Comparison of chlorophyll a
(mg/g
) concentration between plants exposed to high
light intensity vs. low light intensity
Exposed to HIGH light intensity

1 0.802 0.304 0.9368 0.320 1.2573

8

2 0.550 0.220 0.6393 0.246 0.8855

4

3 0.757 0.299 0.8810 0.330 1.2111

4

Exposed to LOW light intensity

1 0.497 0.314 0.5467 0.486 1.0329

5

2 1.032 0.456 1.1880 0.561 1.7488

3

3 1.035 0.444 1.1950 0.532 1.7270

4
FIGURE 5. Comparison of chlorophyll b
concentration between plants exposed to high
light intensity vs. low light intensity
Plants exposed to high light intensity
exhibited higher absorbance both at 663 nm and
645 nm. The chlorophyll a, b, and total
chlorophyll content, however, is generally higher
in plants exposed to low light intensity (Figure 4,
5, and 6, respectively). Conversely, the
 10 1 (Y) 1 (R) 2 (R) 2 (R) 3 (R) 1 (R)

12 1 (Y) 2 (R) 2 (R) 2 (R) 3 (R) 2 (R)

15 1 (Y) 2 (R) 2 (R) 2 (R) 4 (R) 2 (R)

18 1 (Y) 2 (R) 2 (R) 3 (R) 4 (R) 3 (R)

21 1 (Y) 3 (R) 3 (R) 4 (R) 5 (R) 4 (R)

25 1 (Y) 3 (R) 3 (R) 4 (R) 5 (R) 4 (R)

30 1 (Y) 3 (R) 3 (R) 5 (R) 5 (R) 4 (R)

FIGURE 6. Comparison of total chlorophyll

Based on the result, it can be inferred
content between plants exposed to high light
that the blue-cellophane-wrapped tube (A)
intensity vs. low light intensity
exhibited the fastest rate of photosynthesis. This
is succeeded by the red-cellophane-wrapped
tube (B) and green-cellophane-wrapped tube
Part 2. Measuring photosynthesis under
(C).
varying wavelengths of light
The first two test tubes (Test tube A, and
In analyzing the effects of different light
Test Tube B) are different from the rest because
wavelengths on photosynthesis, Hydrilla
certain factors in this experiment were removed.
verticillata was the species observed. As
Test Tube A is significant in the experiment
mentioned, the hydrilla tubes contain: a) no
because it is the Control setup. This means that
baking soda + no wrap, b) with baking soda +
it contains phenol red but is absent of baking
foil-wrapped, c) with baking soda + no wrap, d)
soda and is not wrapped in aluminum foil. By
with baking soda + red cellophane wrap, e) with
removing the baking soda from the control
baking soda + blue cellophane wrap, and f) with
setup, there will no longer be another source of
baking soda + green cellophane wrap.
carbon dioxide in the test tube. Carbon dioxide
Table 2. Solution color changes under varying coming from baking soda can cause the
wavelengths of light. (Yellow as Y, Red as R; induction of photosynthesis when light is
Color intensity = “1” as lightest to “5” as darkest) present. Test Tube B also has significance in the
experiment because it is wrapped in aluminum
Time Tube Tube Tube Tube Tube Tube
(min) A B C D E F foil, which leads to the absence of light
0 1 (Y) 1 (R) 1 (R) 1 (R) 1 (R) 1 (R) interacting with the solution.
2 1 (Y) 1 (R) 1 (R) 1 (R) 1 (R) 1 (R)

4 1 (Y) 1 (R) 1 (R) 1 (R) 1 (R) 1 (R)

Part 3. The Role of chlorophyll and light in
6 1 (Y) 1 (R) 1 (R) 1 (R) 1 (R) 1 (R)
starch synthesis
8 1 (Y) 1 (R) 1 (R) 1 (R) 2 (R) 1 (R)
 Each setup (setups A-H) contained four light
leaf discs in a petri dish varying in color and light H. glucose negative pale white in middle,
solution, white slightly darker yellow
setup. Setups A-D were submerged in distilled discs, in dark edges due to extra
water, while setups E-H were submerged in 5% bleaching and addition o
iodine solution
glucose solution. On the other hand, setups A,
C, E, and G contained discs that were exposed
As seen in Table 3, all setups left in the
to a bright light setup, and setups B, D, F, and H
dark without any sunlight or artificial light
contained discs that were hidden in the dark.
displayed a negative starch test, identified by the
Table 3 shows the results of the starch test after
lack of dark blue-black spots on the discs after
leaving the petri dishes containing the leaf discs
being treated with iodine solution for one minute.
in their assigned setups for 72 hours.
Green discs in the dark barely had any color
change except for the slight bleaching of the leaf
Table 3. Treatments and starch test results and disc margins. White discs remained white in the
descriptions
middle with a slight yellow coloration at the
Treatments Starch Test Color and its intensity;
margins due to the addition of iodine solution.
(positive/negative; distribution of coloration
indicate location of on the leaf Green
discs discs in the bright light setup showcased
starch on leaf dark brown pigmentation in glucose solution and
discs)
distilled water, while white discs in bright light
A. distilled positive dark brown pigment,
water, green outer edge is darker,
showed andlight brown pigmentation in both glucose
discs, in bright center part is lighter
and distilled water placements. Green discs
light
B. distilled negative dark green in placed
middle in
ofbright light exhibited positive results in
water, green disc, slightly white
the starch test.
discs, in dark around edges with yellow
coloration due to iodine
solution
C. distilled negative light brown pigment,
water, white similar pigments4 in the
| Discussion
discs, in bright outer and inner part
light Part 1. Effects of light intensity on leaf
D. distilled negative pale white in the middle,
chlorophyll concentrations
water, white darker white along
discs, in dark edges; entire disc slightly
yellow colored due to Chlorophylls are specialized light-
iodine solution absorbing pigments found in the chloroplast that
E. glucose positive dark brown pigment,
solution, green powers
outer part is darker thanthe process of photosynthesis (Taiz &
discs, in bright the inner part Zeiger, 2010). Chlorophyll has two main types,
light
namely Chlorophyll a (Chla) and Chlorophyll b
F. glucose negative dark green in middle,
solution, green edges slightly (Chlb),
bleached
in which the former is considered as the
discs, in dark and few bleached spots
primary photosynthetic pigment while the latter
going inwards
G. glucose negative is an accessory pigment. Both chlorophyll types
light brown pigment,
solution, white outer and inner are
part have
similar but differ in their structure and
discs, in bright similar pigments
absorption spectra. Chla is abundant in most Therefore, plants exposed to low light intensity
photosynthetic organisms as it acts as a primary synthesized more chlorophyll a & b to enhance
electron donor in the Electron transport chain the light-harvesting complex and maximize the
within the reaction center (Martin, 2019; Croce & light absorption for photosynthesis. A study
van Amerongen, 2014). It absorbs violet (380- conducted by Tran (2018) had similar results
450 nm) and red light (620 –750nm) with peak at where their 4-week samples of Fraxinus latifolia
662 nm. While Chlb, which is bound to Lhc seedlings treated with 50% light exposure had a
proteins, is an antenna and accessory pigment significantly higher total chlorophyll content than
that widens the absorption spectra to absorb those treated with 100% light exposure.
more solar energy from higher frequencies and
Chlorophyll a/b ratios are used to
transfer this energy to Chla for conversion to
determine and quantify species’ shade
chemical energy (Croft & Chen, 2018;
tolerance. These species have lower chlorophyll
Voitsekhovskaja & Tyutereva, 2015). Chlb
a/b ratio (meaning a higher b/a ratio) than
mostly absorbs blue (400-525 nm) and yellow
species with normal light conditions (Yamazaki,
light (565-590 nm) but its peak could be
2005). The chlorophyll a/b ratio of the leaves
observed at 646 nm. Since the pigment granules
exposed to low light intensity was lower than
are the sole light absorbers of the plant, only the
those exposed to high light intensity. It meant
wavelength they take in are used during
that the Chlb concentration of the leaves in the
photosynthesis. This is somehow beneficial to
low light intensity setup was increased while the
plants since the visible light absorbed by the
Chla concentration remained unchanged. As
pigment granules contain the most appropriate
discussed previously, shade-tolerant plants have
amount of energy to excite the electrons needed
higher Chlb concentrations to maximize the light
to initiate photosynthesis without the risk of
absorbed by the plant by expanding its
breaking the DNA content of the plant cells.
absorption spectra (enabling absorption of blue
From all points taken, both chlorophyll a and
light). The present study had similar results to a
chlorophyll b thus influence the photosynthetic
study by Lichtenthaler et al. (2007) where sun
capacity of a plant species.
leaves of the studied deciduous tree species
In the present study, leaves exposed to had a higher Chl a/b ratio than that of the shade
low light intensity (shade setup) were observed leaves.
to have higher concentrations of chlorophyll a, b,
and total chlorophyll content than leaves
Part 2. Measuring photosynthesis under
exposed to high light intensity (Figure 4-6).
varying wavelengths of light
Higher total chlorophyll content, especially
concentration of Chlb, is an adaptive response Photosynthetic activity in light is
for shade-tolerant plants as it maximizes the dependent on the wavelength. Specifically, the
total absorption of the plant with low light active radiation ranges from 400 to 700 nm while
available (Beneregama & Goto, 2010). the unimportant wavelength is less than 400 nm
or more than 700 nm due to the low photon The 8 leaf disc setups were placed in
emission of carbon dioxide assimilation. two different environments in order to observe
their starch content after 72 hours. Since 4 of
The plant choice of this experiment was
the setups do not get light, they would be unable
the water thyme, or Hydrilla verticillata. It is an
to continue with photosynthesis, as a major step
aquatic plant that is usually used when testing
of photosynthesis includes a light reaction which
photosynthesis. A possible role of aquatic
clearly requires light (Johnson, 2016).
plants under climate change based on the
results would be to isolate the leaf’s function Two of these disc setups were placed in
from the other parts of a terrestrial plant. Aquatic a 5% glucose solution, which should allow them
plants have been adapted to an environment to absorb the glucose that can be converted into
that contains less light for aquatic plants to starch. One of the disc setups contained white
absorb, thus giving it a higher chance of discs, which contained no active chloroplasts,
survival. The lack of cuticles on the leaves of therefore unable to proceed with photosynthesis
aquatic plants makes it easier for the absorption (Yamasaki et al, 2014); while one of the setups
of carbon dioxide in the solution. had green discs and should have been able to
show starch spots after the addition of iodine but
In green leaves, there is a high
was unable to due to lack of time in the hot
absorptance for blue (400-495 nm), then red
water bath in 70% ethanol.
(620-750 nm). The lowest rate of photosynthesis
was exhibited by the green-cellophane-wrapped The green discs submerged in distilled
tube. This can be explained due to the water in the dark did not show any major color
appearance of the plant, which is green; changes other than slight bleaching around the
indicative of its low absorptance of green light. leaf disc margins, which shows that the discs
Hence, the low photosynthetic efficiency. were not boiled long enough in 70% ethanol,
since the chlorophylls were not completely
dissolved and the green color still remained
Part 3. The Role of chlorophyll and light in (BBC, 2022).
starch synthesis
Based on the experiment conducted for
Starch reacts effortlessly with iodine light set up, some of the specimens were unable
(Etxeberria et al, nd), which makes it the specific to produce starch which indicates that the
stain in the identification of starch in leaves and photosynthesis did not successfully operate. It is
should produce a black or very dark blue color known that light may increase the rate of
that indicates where the starch is present on the photosynthesis, however this is not the case for
leaf (Moran, nd). Starch reacts effortlessly with some plant species. In various species, too
iodine. In the absence of the dark blue-black much light may damage the photosynthetic
spots, then the consensus is that there is no machinery and it may cause cell death. It also
starch detected on the leaf.
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%20long-chain%20polymer%20of%20glucose
%20molecules,wheat%2C%20to%20store
%20the%20glucose%20made%20by
%20photosynthesis.?
Sunny setup: Replicate 2
msclkid=84fe0444a8e511ec800ecf4753622788
Chlorophyll a
BBC (2022). Photosynthesis. Retrieved from 25
¿ 12.7 ( 0.550 ) −2.69 ( 0.220 ) ×
https://www.bbc.co.uk/bitesize/guides/zpcvbk7/r 1000 × 0.25
evision/3? mg
¿ 6.3932 ×0.1=0.6393
msclkid=ed10046ca8e911ecadb5e5c3e490ed55 g
Chlorophyll b
25
¿ 22.9 ( 0.220 )−4.68 ( 0.550 ) ×
1000 ×0.25
mg
APPENDICES ¿ 2.464 × 0.1=0.2464
g
Sunny setup: Replicate 1
Total Chlorophyll content
Chlorophyll a 25
¿ 20.2 ( 0.220 ) +8.02 ( 0.550 ) ×
25 1000 ×0.25
¿ 12.7 ( 0.802 )−2.69 ( 0.304 ) ×
1000× 0.25
mg
¿ 8.855 ×0.1=0.8855
¿ 9.36764 × 0.1 g
mg 0.6393
¿ 0.9368 Chlorophyll a/b ratios ¿ =2.59
g 0.2464
Chlorophyll b
25
¿ 22.9 ( 0.304 )−4.68 ( 0.802 ) × Sunny setup: Replicate 3
1000 ×0.25
Chlorophyll a
¿ 3.20824 × 0.1 25
¿ 12.7 ( 0.757 )−2.69 ( 0.299 ) ×
mg 1000× 0.25
¿ 0.3208
g mg
¿ 8.80959 ×0.1=0.8810
Total Chlorophyll content g
25
¿ 20.2 ( 0.304 )+ 8.02 ( 0.802 ) ×
1000 ×0.25
Chlorophyll b
¿ 12.57284 × 0.1 25
¿ 22.9 ( 0.299 )−4.68 ( 0.757 ) ×
mg 1000 ×0.25
¿ 1.2573
g mg
¿ 3.30434 × 0.1=0.3304
g

0.9368 Total Chlorophyll content

Chlorophyll a/b ratios ¿ =2.92 25
0.3208 ¿ 20.2 ( 0.299 ) +8.02 ( 0.757 ) ×
1000 ×0.25
 mg mg
¿ 12.11094 ×0.1=1.2111 ¿ 5.61264 × 0.1=0.5613
g g
Total Chlorophyll
25
0.8810 ¿ 20.2 ( 0.456 ) +8.02 ( 1.032 ) ×
Chlorophyll a/b ratios ¿ =2.67 1000 × 0.25
0.3304
mg
¿ 17.48784 × 0.1=1.7488
g
Shade setup: Replicate 1
1.1880
Chlorophyll a Chlorophyll ratio = ¿ ¿ 2.12
0.5613
25
¿ 12.7 ( 0.497 )−2.69 ( 0.314 ) ×
1000 ×0.25
mg
¿ 5.46724 × 0.1=0.5467
g Shade setup: Replicate 3

Chlorophyll b Chlorophyll a
25 25
¿ 22.9 ( 0.314 )−4.68 ( 0.497 ) × ¿ 12.7 ( 1.035 )−2.69 ( 0.444 ) ×
1000 × 0.25 1000× 0.25
mg mg
¿ 4.86464 ×0.1=0.4865 ¿ 11.95014 ×0.1=1.1950
g g
Total Chlorophyll Chlorophyll b
25 25
¿ 20.2 ( 0.314 )+ 8.02 ( 0.497 ) × ¿ 22.9 ( 0.444 )−4.68 ( 1.035 ) ×
1000 × 0.25 1000 ×0.25
mg mg
¿ 10.32874 × 0.1=1.0329 ¿ 5.3238 ×0.1=0.5324
g g
Total Chlorophyll
25
0.5467 ¿ 20.2 ( 0.444 )+ 8.02 ( 1.035 ) ×
Chlorophyll ratio = ¿ ¿ 1.12 1000 ×0.25
0.4865
mg
¿ 17.2695 ×0.1=1.7266
g
Shade setup: Replicate 2

Chlorophyll a
25 1.1950
¿ 12.7 ( 1.032 )−2.69 ( 0.456 ) × Chlorophyll a/b ratio ¿ =2.24
1000 × 0.25 0.5324

mg
¿ 11.87976 × 0.1=1.1880
g
Chlorophyll b Part 2. Measuring photosynthesis under varying
25 wavelengths of light: Images
¿ 22.9 ( 0.456 ) −4.68 ( 1.032 ) ×
1000× 0.25