Department of Biological Sciences

College of Arts and Sciences

Visayas State University
Visca, Baybay City, Leyte 6521-A Philippines

MarB 15 - GENERAL PHYSIOLOGY

PHOTOSYNTHESIS:
FLOATING LEAF DISK ASSAY FOR QUANTITATIVE INVESTIGATION OF
PHOTOSYNTHESIS
Name : Eugene L. Codis Date Performed: November 26, 2019
Lab Sched: TTH (07:00-10:00 AM) Date Submitted: December 10, 2019

RESULTS
I. Experiment 1: Effect of CO2 on Photosynthethic rate of the Leaf Disks
Table 1. Leaf disk infiltrated with NAHCO3 and dH2O and were immersed in bicarbonate
solution

Time No. of floating leaf disk in bicarbonate solution

(minutes) NAHCO3 infiltrated Water infiltrated
3 0 0
6 0 0
9 0 0
12 0 0
15 0 0
18 0 0
21 0 0
24 0 0
27 0 0
30 0 0
33 0 0
36 0 0
39 0 1
42 0 0
45 0 0
48 0 0
51 0 0
54 0 0
57 0 0
60 0 0
 NaHCO3 infiltrated Leaf Disks Water infiltrated Leaf Disks

1.2
1
No. of floating leaf disks

0.8

0.6

0.4

0.2
00 00 00 00 00 00 00 00 00 00 00 00 0 00 00 00 00 00 00 00
0
3 6 9 12 15 18 21 24 27 30 33 36 39 42 45 48 51 54 57 60
TIme (minutes)

Figure 1. Effect of bicarbonate solution in photosynthesis.

Table 2. Leaf disk infiltrated with NaHCO3 and dH2O and were immersed in distilled
water with 5cm depth as (control).

Time No. of floating leaf disk in water solution

(minutes) NaHCO3 infiltrated Water infiltrated
3 0 0
6 0 1
9 0 0
12 1 0
15 0 0
18 0 0
21 0 0
24 0 1
27 0 0
30 0 0
33 0 0
36 0 0
39 0 0
42 0 0
45 0 0
48 0 0
51 0 0
54 0 0
57 0 0
60 0 0
 NaHCO3 Infiltrated Disks Water Infiltrated Disks

1.2
1 1 1
Number of floating leaf disks

0.8

0.6

0.4

0.2
00 0 00 0 00 00 00 0 00 00 00 00 00 00 00 00 00 00 00 00
0
3 6 9 12 15 18 21 24 27 30 33 36 39 42 45 48 51 54 57 60
Time (minutes)

Figure 2. Effect of distilled water in photosynthesis (control).

II. Experiment 2: Effect of Light Intensity on Photosynthesis

Table 3. Leaf disks exposed to varying light intensity by increasing distance from water
surface.

Time No. of floating leaf disk exposed to different distances from light
(minutes) source
5cm 10cm 15cm
R1 R2 R1 R2 R1 R2
3 0 0 0 0 0 0
6 0 0 0 0 0 0
9 0 0 0 0 0 0
12 0 0 0 0 0 0
15 0 0 0 0 0 0
18 0 0 0 0 0 0
21 0 0 0 0 0 0
24 0 0 0 0 0 0
27 0 0 0 0 0 0
30 0 0 0 0 0 0
33 0 0 0 0 0 0
36 0 0 0 0 0 0
39 0 0 0 0 0 0
42 0 0 0
45 0 0 0
48 0 0 0
51 0 0 0
54 0 0 0
57 0 0 0
 60 0 0 0

III. Experiment 3: Effect of Chlorophyll on Photosynthesis

Table 4. Leaf disks from green and variegated Hibiscus rosa-sinensis infiltrated with
NaHCO3 and immersed in distilled water with 5cm depth.

Time No. of floating leaf disk in water solution

(minutes) Green Variegated
R1 R2 R3 R1 R2 R3
3 0 0 0 0 0 0
6 0 0 0 0 0 0
9 0 0 0 0 0 0
12 0 0 0 0 0 0
15 0 0 0 0 0 0
18 0 0 0 0 0 0
21 0 0 0 0 0 0
24 0 0 0 0 0 0
27 0 0 0 0 0 0
30 0 0 0 0 0 0
33 0 0 0 0 0 0
36 0 0 0 0 0 0
39 0 0 0 0 1 1
42 0 1 0 0 0 0
45 0 0 2 0 1 0
 Green 1 Green 2 Green 3 Variegated1 Variegated2 Variegated3

2.5

2
2
Number of floating leaf disks

1.5

11 1 1
1

0.5

000000 000000 000000 000000 000000 000000 000000 000000 000000 000000 000000 000000 0000 0 0000 00 0 0
0
3 6 9 12 15 18 21 24 27 30 33 36 39 42 45
Time (minutes)

Figure 3. Effect of Chlorophyll of Photosynthesis

DISCUSSION
Leaves from Imperata cylindrica and Hibiscus rosa-sinensis have been used as test
samples for the floating leaf disk assay. Leaf disks have been infiltrated withNaHCO3 as
a source of carbon dioxide and dH2O soap solution. By removing all the trapped gas
within the leaf disks and replacing it with either soap solution, NaHCO3 or dH2O, the
penetration was achieved through a vacuum.

Figure 1 illustrates the effect of CO2 on Experiment 1 photosynthetic rate of leaf

disks. The leaf disks came in two groups: NaHCO3 infiltrated and water infiltrated. Both
were immersed in 5 cm deep with a solution of sodium bicarbonate. During the 39th
minute of immersion, only one leaf disk floated throughout an hour. On the other hand,
the leaf disks shown in Figure 2 are the same as the first set-up but are then immersed in
water 5 cm deep. During an hour of immersion, three-leaf disks floated. During the 6th,
12th and 24th minute of immersion, they drifted. This test could have a flaw allowing a
plant to be exposed to a CO2 stream, so H2O should allow the cycle of photosynthesis to
take place, and the leaf disks should have floated as a symbol that the process took place.

The influence of light intensity on photosynthesis was calculated in the second

experiment by exposing the leaf disks to differing light intensity by growing distance
from the surface of the water. In photosynthesis, light intensity also shares the
importance of CO2 and H2O. The system would be shut down without it. During an hour,
no leaf disks are found. Nevertheless, researchers observed that the surface of leaf disks
contained some development of bubbles. The light source formed 33 bubbles in 10
leaves at 10 cm. Thirteen bubbles were formed in four leaves at a distance of 5 cm and
eight bubbles at a distance of 15 cm. The oxygen produced in the process of
photosynthesis is the bubbles found on each leaf disk's surface. Oxygen on the ground
could not be sufficient to allow the leaf disks to move, which is why it stays at the base.
There is no significant trend as to whether this light intensity raises photosynthetic levels
at an average distance, but the photosynthetic frequency could be higher due to
numerous bubble shapes like this in the 10 cm wavelength set-up.

In Experiment 3, the influence of chlorophyll on photosynthesis was calculated by

infiltrating the green and variegated leaf disks of Hibiscus rosa-sinensis with NaHCO3
and immersing them in 5 cm deep distilled water. Variegated leaf disks have less
chlorophyll in them because they are patterned in a secondary color other than green that
may decrease the photosynthesis speed. This experiment may also have an error because
there has been no significant evidence or observation that the leaf disks from the green
leaves are floating ahead of time or that they are more numerous on floating disks than
the variegated leaves shown in Figure 3.

CONCLUSION
The floating lea disk assay experiment can be used to quantitatively measure the
photosynthetic rate of plants. Photosynthesis is the most basic process occurring on Earth
every day, but to fulfill its function it still has complex and essential processes. Powered
by light energy from the sun, carbon dioxide and water are required for photosynthesis.
Light energy is absorbed by light-absorbing pigments in plant leaves such as chlorophyll.
Many variables can influence levels of photosynthesis, such as carbon dioxide
concentration, light intensity, and chlorophyll content.

ANSWERS TO QUESTIONS
1. Why does the leaf disk sink after infiltration and rise during photosynthesis?
Leaf disks normally float, however when the air spaces are infiltrated with carbon
dioxide, the overall density of the lead disk increases and the leaf disk sinks. When
sodium bicarbonate is added to the water, the bicarbonate ion acts as a carbon source
for photosynthesis causing the leaf disks to sink.(https://www.nthurston.k12)
2. Based on your data, explain briefly the effects of the following factors on
photosynthesis:

a. Light intensity

Light intensity is one of the factors affecting the rate of photosynthesis. Light
intensity directly affects the light-dependent reaction in photosynthesis and indirectly
affects the light-independent reaction. As light intensity increases (distance between
 lamp and plant decreases) the volume of oxygen (or the rate of bubble production)
increases. This indicates that the rate of photosynthesis increases with light intensity.
However, at sufficiently high levels of light intensity, the rate oxygen evolution
remains constant. (brilliantbiologystudent.weebly.com, 2015)

b. Carbon dioxide

An increase in the amount of CO2 well increase the rate of photosynthesis,

Carbon dioxide concentration will directly affect the rate of photosynthesis as it is
used in the photosynthesis reaction.Increased amount of CO2 will increase the rate of
photosynthesis to a certain limit, after which a further increase in its amount will no
longer increase the rate any further. (Henry Williams, 2016)

c. Chlorophyll

The amount of chlorophyll also affects the rate of photosynthesis: plants in

lighting conditions unfavourable for photosynthesis may synthesise more chlorophyll,
to absorb the light required. the effects of some plant diseases affect the amount of
chlorophyll, and therefore the ability of a plant to photosynthesise.

3. What is the importance of ET50 in the leaf disk floating technique on studying
photosynthesis?
The ET50 is the time it takes for 50% of the leaf disks to float and is a good
indicator of the rate of photosynthesis. It was hypothesized that if the concentration
of carbon dioxide was increased, then the rate at which photosynthesis occurs will
also increase. (Robinson-Brown, 2017)

REFERENCES
nthurston.k12.wa.us/cms/lib/WA01001371/Centricity/Domain/2015/Leaf%20Disk%20P
hotosynthesis%20Lab%202013.pdf
http://brilliantbiologystudent.weebly.com/effect-of-light-intensity.html
https://www.quora.com/How-does-the-rate-of-photosynthesis-relate-to-carbon-dioxide-le
vels
https://bam.files.bbci.co.uk/bam/live/content/zgcvdxs/large
https://scholarcommons.sc.edu/cgi/viewcontent.cgi?article=1156&context=jscas