FACULTY OF APPLIED SCIENCE

BACHELOR OF SCIENCE (HONOURS) BIOLOGY

CAMPUS KUALA PILAH, NEGERI SEMBILAN

BIO611 PLANT PHYSIOLOGY

EXPERIMENT 2: STOMATAL DISTRIBUTION ON LEAVES OF
SEVERAL SELECTED PLANTS

PREPARED BY:

NAME STUDENT ID GROUP

AHMAD ANIQ HAMKA 2021868692 AS2015A1

BIN AZHAR

NUR NIKMAN BIN 2021899348 AS2015A1

SAMSUDDIN

PREPARED FOR:
MADAM LILI SYAHANI
RUSLI
INTRODUCTION

Plants have small openings called stomata, which are regulated by guard cells and have a
variety of purposes. C3 and C4 plants have different stomata and guard cell sizes and shapes. For
instance, the guard cells of C4 plants resemble dumbbells, whereas the guard cells of C3 plants
are formed like beans. Usually located on the underside of the leaf are stomata. Though less
frequently than on the plant's leaves, it is also visible on the stems, petioles, sepals, and petals of
the plant. One of the primary functions of stomata is to enable transpiration and gas exchange in
plants, which allows the xylem tissues to absorb water from the soil.

Stomatal density is determined by the number of stomata and the size of the epidermal cells.
Stomatal index is the total number of stomata divided by the total number of epidermal cells in
each unit area. It can be used to calculate the numerical distribution of stomata in each leaf.
For this experiment, we will use and observe different plants such as Gardenia sp., Axonopus
compressus and Nymphaea sp.

OBJECTIVES

1. To compare the densities of stomata on abaxial and adaxial leaf surfaces of the same
leaf with relation to transpiration.
2. To compare the densities of stomata of different plants, with relation to transpiration.

MATERIALS

● Fully expanded leaves of dicot, monocot and aquatic plants

● Light microscope
● Microscope slide and cover slip
● Forceps
● Clear nails varnish
● Clear sticky tape
● Digital camera
PROCEDURES

1. Leaves of dicot and monocot were found.

2. The upper and lower surfaces of leaves were identified.
3. A thin layer of clear nail varnish was spread on each surface, upper and lower side of
leaves.
4. Leaves were allowed to dry for 10 to 15 minutes.
5. A strip of clear sticky tape was placed over the nail vanish.
6. The sticky tape was pressed down to make a good connection with the nail varnish.
7. The sticky tape was peeled off. The layer of nail varnish has come off with the tape.
8. The tape was placed on a microscope slide and cover slip was added on it.
9. The slide was labeled with the plant name, upper and lower surface.
10. The slide then placed under the microscope on 40x magnification.
11. The number of stomata were counted 3 times and recorded for every surface of the
leaves.
12. The area of field of view was determined by using a microscopic slide with scale.
13. The result then tabulated and the stomatal density was calculated.
RESULTS

PLANT No. Of No. Of Open No. of Epidermis Stomatal Index (%)

Stomata Stomata
Adaxi Abaxi Adaxi Abaxia Adaxi Abaxial Adaxial Abaxial
al al al l al
Gardeni 2 19 0 7 125 156
a sp.
(dicot) 1 18 1 10 111 140
1.68 11.56
2 15 2 15 120 146

Average 2 17 1 11 119 147

Axonop 15 18 8 18 63 68
us
compres
17 15 7 15 56 67
sus
(monoc 29.62 26.31
ot) 17 26 6 26 43 93

Average 16 20 7 20 54 76

Nympha 51 0 20 0 19 0
ea sp.
(Aquati 30 0 12 0 11 0
c plant)
306.67 0
58 0 5 0 15 0

Average 46 0 12 0 15 0

Table 1: The number of stomata and epidermis on adaxial and abaxial surface of the leaf
 Sample Surface Magnification Number of Stomata
2
Name (Adaxial/Abaxi (Ocular x FOV(cm ) stomata density
al) Objective) (No of
stomata/cm2)
Gardenia sp. Adaxial 400x 1.5904x10-3 2 1258
(dicot)
Abaxial 400x 1.5904x10-3 17 10689
Axonopus Adaxial 400x 1.5904x10-3 16 10060
compressus
(monocot) Abaxial 400x 1.5904x10-3 20 12575

Nymphaea Adaxial 400x 1.5904x10-3 46 28922

sp. (Aquatic
plant) Abaxial 400x 1.5904x10-3 0 0

Table 2: Stomata density on adaxial and abaxial surfaces

DISCUSSION

In this experiment, there were three plants that were observed which were Gardenia sp.,
Axonopus compressus, Nymphaea sp.. We utilized clear nail varnish to observe stomatal
density on both leaves surface, abaxial (lower) and adaxial (upper) under 40x magnification on
a compound microscope. Stomata can be present on both surfaces of the leaves where stomata
number on abaxial is higher than adaxial surface in dicot while stomata number on abaxial have
equal number with the adaxial number.
 In this case, Gardenia sp. is a dicot plant while Axonopus compressus is a monocot plant.
Based on Table 1, average number of stomata in Gardenia sp. for the adaxial surface is 2 while
for the abaxial surface is 17. In Axonopus compressus, the average number of stomata for the
adaxial surface is 16 while for the abaxial surface is 20. In dicot plants, stomata on adaxial
surfaces are lower than abaxial surfaces to prevent water loss during the daytime where there is
the presence of sunlight. If the number of stomata on the adaxial surface is higher it will cause
wilting of the leaves because of high temperature. For the monocot plants, the number of stomata
on both surfaces, adaxial and abaxial is the same because monocotyledon leaves are held
vertically and parallel to sunlight. Thus, both surfaces have the same number of stomata to
receive equal amounts of sunlight. As for the Nymphaea sp. In aquatic plants,the average number
of stomata for the adaxial surface is 46 while the average number of stomata for the abaxial
surface is 0. Usually, stomata are present more abundantly on the abaxial surface when
compared to the adaxial surface of the leaf. In water lilies, since the abaxial surface of leaves are
submerged with the water, stomata are present only on the adaxial surface of the leaves that are
exposed to air in order for the process of gaseous exchange to occur. Other than that, based on
the result in Table 1, the stomatal index for Gardenia sp. on adaxial and abaxial surfaces are 1.68
and 11.56 respectfully. The stomatal index for Axonopus compressus on adaxial and abaxial
surfaces are 29.62 and 26.31.Also the stomatal index for Nymphaea sp. is 306.67 for adaxial
surface and 0 for abaxial surface. The stomatal index formula is the total number stomata is
divided by the total number of epidermis. The importance of stomatal index is to prove that the
distribution of stomata on the leaves surface are consistently the same along the leaves.

In Table 2, the adaxial and abaxial surfaces of Gardenia sp. are 1258 cm2 and 10689 cm2
respectively. The abaxial surface has higher stomatal density than the adaxial surface because
Gardenia sp. is a C3 plant. C3 plant has a large size of guard cells and it is meant to have
higher stomatal density to prevent excessive water loss that may cause growth defect and
dehydration. For Axonopus compressus, the adaxial and abaxial surfaces are 10060 cm2 and 12575
cm2 consecutively. The stomatal density is almost the same on adaxial and abaxial surfaces
because Axonopus compressus is a C4 plant and both surfaces on the leaves are important for
the photosynthesis and transpiration process. For Nymphaea sp. the stomatal density for adaxial
surface is 29822 while on the abaxial surface is 0.This is because Nymphaea sp. is an aquatic
plant.To allow the plant to exchange gases on the adaxial surface, aquatic plants require a large
number of stomata on the upper surface of their leaves.Leaves do not require gas exchange with
the atmosphere on their abaxial surface.To make up for the lack of stomata on the abaxial
surface, leaves have a high number of stomata on their adaxial surface.

CONCLUSION

In a nutshell, the experiment shows that dicot, monocot, and aquatic plants have different
numbers of stomata on their upper and lower leaf surfaces. It made it possible for us to compare
the stomata density in relation to transpiration on the adaxial and abaxial surfaces of the same
leaf. Furthermore, we could assess how different plants' stomatal densities related to
transpiration. Based on the result of the experiment, the lower surfaces of dicot plant has higher
number of stomata than the upper surface while the monocot plant has the equal number of
stomata on both surfaces. In aquatic plants, it has the highest number of stomata compared to the
other two plants. The stomatal densities results also do not contradict with the objectives of this
experiment and the distribution of stomata densities correlates with the transpiration process.

POST-LAB QUESTIONS
1. Compare the stomata density on the upper and lower leaf surface of a leaf.
Stomata density on the upper leaf surface is lower than the lower leaf surface as it helps
to minimize the water loss during transpiration.
2. Contrast the stomata density between dicot and monocot plants.
Stomata density in the monocot plants is higher compared to the dicot plants because
dicotyledon stomata are absent on the lower surface of the leaves but for monocotyledon,
the stomata are present on both sides of the leaves.
3. Calculate the percentage of the stomata with open pore on the upper leaf surface
of the dicot and monocot plant.

Dicot

Open stomata% = 𝑁𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑜𝑝𝑒𝑛 𝑠𝑡𝑜𝑚𝑎𝑡𝑎

𝑇𝑜𝑡𝑎𝑙 𝑛𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑠𝑡𝑜𝑚𝑎𝑡𝑎
x 100%

=½ x 100%
=50%

Monocot

Open stomata% = 𝑁𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑜𝑝𝑒𝑛 𝑠𝑡𝑜𝑚𝑎𝑡𝑎

𝑇𝑜𝑡𝑎𝑙 𝑛𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑠𝑡𝑜𝑚𝑎𝑡𝑎
x 100%

=7/16 x 100%

=43.75%
REFERENCES

Kochhar, S., & Gujral, S. (2020). Plant Physiology: Theory and applications (2nd
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Sakoda, K., Yamori, W., Shimada, T., Sugano, S. S., Hara-Nishimura, I., & Tanaka, Y.
(2020, September 29). Higher stomatal density improves photosynthetic induction and
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