PLANT PHYSIOLOGY (BIO611)

LAB REPORT EXPERIMENT 1: PLANT STRUCTURE AND ANATOMY

LAB REPORT EXPERIMENT 2: STOMATAL DISTRIBUTION ON LEAVES
LAB REPORT EXPERIMENT 3: TRANSPIRATION
LAB REPORT EXPERIMENT 4: PHOTOSYNTHESIS AND CELLULAR
RESPIRATION

STUDENT’S NAME: SITI NURUL AINA BINTI RAZALI

ID NUMBER : 2016579303

GROUP : ED247(8C)

INSTRUCTOR : DR SITI RAFEDAH BINTI ABDUL KARIM

 TITLE: PLANT STRUCTURE AND ANATOMY

OBJECTIVES
1. To study the making of thin cross-sections for plant material such as stems and leafs.
2. To study the staining material using toluidine blue (TBO).

RESULTS AND ANALYSIS

(A) Stem cross-section

This is the image of coleus plant

(plactranthus scutellariodes) under a
compound miscroscope with
magnification of 40x. We can see
clearly the difference between the cells
with TBO stain and without TBO stain.
 Let’s look at the coleus plant cells closely…..

No stain TBO stain

100x

Pith

Xylem

Vascular cambium

Phloem Cortex

Collenchyma
Trichome

400x

Cortex
Collenchyma

Cortex

Epidermis

Two types of
trichomes
Let’s look at the carrot plant cells closely…..

Cortex

Phloem

Cambium

Pith ray

Xylem

(B) Leaf cross-section

 Let’s look at the stoma cells on a leaf closely…..

DISCUSSION

In this experiment, we studied on the stem and leaf cross-section under a microscope.

Based on the first experiment, the plant material used is Plectranthus scutellarioides (coleus) and

Daucus carota subsp. sativus (carrot). Firstly, we observed coleus and carrot stem structure. Both

coleus and carrot stem structure have cortex, phloem, xylem and vascular cambium. Cortex is a

multi-layered well developed and made from oval parenchymatous cells with intercellular spaces.

The intercellular spaces usually help in gaseous exchanges and storage for starch. Cortex also

helps in mechanical support to the roots. Xylem and phloem are complex tissues that perform

transportation of food and water in a plant. They are the vascular tissues of the plant and together

form a vascular bundle.

Xylem is a vascular tissue that transports water and dissolved minerals nutrients from roots

to various parts of the plant. It is responsible for replacing water lost through transpiration and

photosynthesis. While phloem responsible for transporting sugars, proteins and other organic

molecules from the green parts of the plant to the rest of the plant. The vascular cambium

generates the xylem and phloem of the vascular system, which are used for transport and support.

It is a single layer of meristematic cells that undergoes an expansion during the transition from

primary to secondary growth.

 Secondly, we observed that both coleus and carrot stem cross-section have a few different

structures which are coleus have epidermis, trichomes, pith and also ground tissue that is

collenchyma cells. While carrot only have medulla rays, instead of cortex, xylem, phloem and

vascular cambium. In coleus, epidermis is the outermost layer of the leaf. It consists of the upper

and lower epidermis, which are present on either side of the leaf. It aids in the regulation of gas

exchange via stomata. Two guard cells surround each stoma, regulating its opening and closing.

Trichomes are the small, hair-like growths which are found in coleus and also many types

of plant. Trichomes emerge from the surface of buds, stalks, stems and leaves of a plant. It helps

to reduce evaporation by protecting the plant from wind and heat. In many cases, trichomes protect

plants from herbivorous insects that may want to feed on them. Pith of a plant is composed of soft,

spongy parenchyma cells, which store and transport nutrients throughout the plant. In coleus, pith

is located in the centre of the stem.

On the other hand, collenchyma cells are elongated cells with irregularly thick cell walls that

provide support and structure of the plant. Their thick cell walls are composed of the compounds

cellulose and pectin. These cells are often found under the epidermis or the outer layer of cells in

young stems and in leaf veins. In carrot cross section, it contains a structure called medulla rays,

also known as vascular rays or pith rays. It is essential for the radial conduction of the water,

minerals and other organic substances. It also transports the substances from centre to periphery.

For the second experiment, we studied on the leaf cross-section. Firstly, we prepared a thin

specimen of the leaf section to be observed under a microscope. We added a few drops of iodine

to the leaf specimen to observe the cells and tissues of the leaf clearly. We prepared two different

slides of the leaf specimen, the first was stained by using iodine and the second one without

staining, which is only dipped into distilled water. We can see clearly under the microscope the

structure of the leaf which contains palisade mesophyll, spongy mesophyll, cuticle which is located

outside the epidermis and protects leaf against water loss and also stomata, that aids in the

process of transpiration.
CONCLUSIONS

In plant anatomy, structure of the cells is categorized broadly into three tissue systems

which are the epidermis, ground tissue and vascular tissue. Epidermis is cells forming the outer

surface of the leaves and of the young plant body. While vascular tissue is a complex conducting

tissue, formed more than one cell type. The primary components of vascular tissue are the xylem

and phloem. These two tissues transport fluids and nutrients internally. Besides, ground tissue is

less differentiated than the other plant tissues. It is manufactures nutrients by photosynthesis and

store reserve nutrients. Ground tissue can be divided into three types based on the nature of cell

walls, which are collenchyma, parenchyma and sclerenchyma. Based on the experiment of stem

cross-section, both coleus and carrot plant have a few different structures which are coleus have

epidermis, trichomes, pith and also ground tissue that is collenchyma cells. While carrot only have

medulla rays, instead of cortex, xylem, phloem and vascular cambium. While in leaf cross-section,

it consists of palisade mesophyll, spongy mesophyll, stomata and also cuticle.

TITLE: STOMATAL DISTRI BUTION ON LEAVES

OBJECTIVES

1. To study the structure of stomata on a leaf peel.

2. To study the stomatal distribution on the upper and lower leaf surfaces.

RESULTS AND ANALYSIS

This is the image of balsam plant

(Impatiens balsamina) under a
compound miscroscope with
magnification of 100x. We can see
Cell
clearly the structures of the stomata and
the guard cells. Cell walls

Cytoplasm

Guard cells

Stomata

The visible stomata appear under

the microscope, N= 62

The highest stomata counts were 62. It is observed

under a microscope with a magnification of 40x

Area = ∏ x (D/2)2 Stomatal distribution

= ∏ x (135 µm)2 = number of stomata/area

= 57 255 µm2 = 62/0.0573 mm2

= 0.0573 mm2 = 1083 mm2

DISCUSSION

Based on the experiment, the plant material used is balsam (Impatiens balsamina). We

plucked one fresh balsam leaf and take the peel from the upper surface by using the forceps. The

peel then was placed into a watch glass containing distilled water to allows the cells to remain

turgid. Another peel from lower surface of the leaf was repeated in another watch glass. A few

drops of Safranin solution were placed into both watch glass in the experiment. The solution was

used for staining plant cells and allow rapid detection of a plant’s vascular tissue.

After transferring these two leaf peels on the glass slides by using brush, we cut a small

piece from each peel. Then, we put one drop of glycerine on both slides to preserve the leaves

and gives them a more natural appearance appropriate for dry arrangements. Next, we took a

cover slip and place it gently on the peel with the help of needle to avoid the peel torn. Finally, we

count the number of stomata in the peels of both upper and lower epidermis of the leaf. We

observed that the number of stomata is greater in the lower epidermis and fewer are present in

upper epidermis of the leaf.

Under the microscope, the stomata are counted when we found an area with high density

of stomata that followed consistency rule with only partially visible along the edge of the field or

view. This method is repeated at least three times for each leaves to build up the sample size.

Once we get three highest of stomata counts, we can get the data point from the observation. In

the experiment, it showed three highest density of stomatal distribution of the leaf which are N=62,

N=59 and N=51 respectively. Next we calculated the stomatal distribution by using the formula

given which is:

Area = ∏ x (D/2)2
 In this experiment, there are some precaution steps that we need to take note. Firstly, when

we choose a sample of plant, the epidermal peel should be taken from a freshly plucked leaf and

then take the epidermal from the lower surface of a leaf, as it has more stomata. On the other hand,

we have to be careful when pulled out the thin membrane of the peel and avoid the curling of the

peel. Next, we must always use a clean glass slide to prevent contamination and unwanted

substances in the sample. Last but not least, always use a brush to transfer the peel from watch

glass to the slide.

CONCLUSION

On leaf structure, stomata are minute pores found on the epidermis of leaves and young

shoots of plants that are used to control exchange of gases. The pore is surrounded by a pair of

specialised cells called the guard cells that are responsible in regulating the size of the opening.

Water is released through the stomata into the atmosphere in the form of water vapour through the

process called transpiration. Besides, the exchange of oxygen and carbon dioxide in the leaf also

occurs through the stomata.

The distribution of stomata varies between monocots and dicots, between plant species

and between the upper and lower side of the leaves on a plant. Stomata are found more on plant

surfaces thriving under higher light, lower atmospheric carbon dioxide concentrations and in moist

environments. In this experiment, the lower surface of a dicot leaf like balsam plant has a greater

number of stomata while in a monocot leaf, they are more or less equal on both surfaces. However,

in most of the floating plants such as water lilies, stomata are found only on the upper epidermis.
TITLE: TRANSPIRATION – PLANT ABSORBS WATER THROUGH ROOTS

OBJECTIVES
1. To understand how water moves from roots to the leaves.
2. To understand the role of transpiration in the transport of water within a plant.

RESULTS AND ANALYSIS

Day 1

Day 2
 Test tube Setup Level of water Condition of the
after 1 day Purslane plant
A Test tube containing only water and Remain the same No change
rubber cork.
B Purslane plant roots immersed in the Decreased Remain healthy
test tube containing water and mustard
oil.
C Purslane plant roots immersed in the Decreased Remains healthy,
test tube containing water, mustard oil stem and leaves
and red ink. have turned red.
D Purslane plant roots not immersed in the Remains the same Has shrivelled
test tube containing water .

DISCUSSION

In this experiment, the plant material used is purslane plant (Portulaca oleracea) to show

how different effects such as the presence of mustard oil and red ink affect transpiration process.

The results of transpiration under normal room conditions on the plant showed the effectiveness

of the plant absorbs the water. There are 4 test tubes used to differentiate the effects of the

absorption of water. Test tube A, B and C are filled with three-fourths water while test tube D is

added with a few drops of water. A rubber cork is sealed on the mouth of test tube A. Then, three

purslane plants with roots are placed in the test tube B, C and D.

The purslane plant must be washed first to get rid of the soil particles, then the roots must

be intact with water in these test tubes except for test tube D which contain less water. Mustard oil

is added to bot test tube B and C, which acts to reduce transpiration rate by blocking stomata and

intercellular spaces. Then, test tube C is added by a few drops of ink. After 2 days, the water level

and condition of purslane plant of each test tube was observed. In test tube A, there is no change

in the water level proving that there is no absorption of water.

 While in test tube B and C, there is a decrease in the water level confirming that roots

absorb water which is lost through transpiration. In test tube C, there is a rise of sap, that is water

along with red ink is absorbed and transported to all parts of the plant. It is confirmed by the change

in the colour of the stems and leaves. In test tube D, as the roots are not in contact with water, the

water is not absorbed by the plant resulting in its death.

There are some precautions while carried out this experiment. Firstly, always ensure that

the roots of plant are devoid of any soil particles. Next, we must ensure that the roots of the plant

are completely immersed in the water for the transpiration process. Then, we need to mark the

level of water in all the test tubes using a marker so that we can see the differences after 2 days

of the experiment.

CONCLUSION

The experiment of transpiration consists of testing how plants can evaporate water from

their roots and out of the leaves through different condition. Based on the observation between

four test tubes containing different element, purslane plant can absorb water by the presence of

water and mustard oil. Mustard oil helps to reduce transpiration rate by blocking stomata and

intercellular spaces. The presence of oil on the water surface will blocks sunlight and prevents air

from reaching the plants that live in the water. The primary way for plants to absorb water is through

their roots. The water in the soil gets drawn into the roots because of a lower concentration of water

there. Once it is in the roots, the plants use its transport system to relocate the water to the leaves,

buds and flowers. To conclude, plant absorbs water only through their roots and transport it to

other parts of the plant.

TITLE: PHOTOSYNTHESIS AND CELLULAR RESPIRATION

OBJECTIVES
1. To observe the effect of the color of bromothymol blue solution after photosynthesis of
plant.
2. To study the effect of the color of bromothymol blue solution in cellular respiration.

RESULTS AND ANALYSIS

(A) Photosynthesis

The results
after 2 hours

Test tube 1 2 3 4 5
(constant)
Light treatment Dark Light Dark Light Light

Test tube treatment Elodea + Elodea+ Leaf + CO2 Leaf+ CO2 No plant
CO2 CO2
Initial color Green Green Green Green Green

Final color Unchanged Blue Unchanged Blue Green

 (B) Cellular respiration

The results
after 24 hours

Test tube Color at the beginning of Color changes after 24 hours

experiment
A Blue Yellow green

B Blue No change

DISCUSSION

During the photosynthesis, the light intensity will increase when the rate of photosynthesis

increase. Based on this experiment, there were two leaves used which are Elodea plant and some

random leaves from bushes. We prepared five test tubes containing bromothymol blue solution.

Bromothymol blue is used as an indicator for the presence of carbon dioxide in a solution. When

carbon dioxide dissolves in water, carbonic acid is formed. A bromothymol blue solution acidified

to pH 6.0 by an addition of carbon dioxide produces a yellow color. The blue color is restored when

the carbon dioxide is removed and the pH becomes higher than 7.6.

The bromothymol blue is blue when it is free of dissolved CO2 gas, and is yellow when CO2

is dissolved in the solution. The process actually causing the shift in color is a pH change (change

in the H+ concentration of the solution). Carbon dioxide reacts with water in the following way:

CO2 + H2O H2CO3 H+ + HCO3-

 A plant in the dark respires but does not carry out all stages of photosynthesis. Because

CO2 is being generated by respiration but not consumed by photosynthesis, there is a net

accumulation of CO2. As the concentration of CO2 increases in solution, the above reaction

increases the H+ ion concentration, thereby lowering the pH. A plant in the light can carry on all

stages of photosynthesis. The CO2 used in photosynthesis per unit time is much greater than the

amount generated by respiration over the same time period (about 10-30 times greater). Therefore,

in the presence of light, Elodea will take up CO2, causing the concentration of CO2 in its

surroundings to drop. This causes a decrease in the H+ ion concentration through the reversible

reactions above and, therefore, the pH will increase. The following reaction takes place:

HCO3- + H+ → H2CO3 → H2O + CO2

In this experiment, there are some precautions that we had to be aware. Firstly, be careful

not to swallow any bromothymol solution or splash it in your face because it is toxic if swallowed.

Secondly, dispose bromothymol blue solution once complete the observation. This is done by

pouring it down the drain, and make sure to rinse the sink with plenty of water so we do not leave

any residue. Next, never touch or taste any chemicals and always handle glassware carefully. Last

but not least, be careful with the bromothymol blue solution since it can cause stains on clothing

and if it gets into eyes, immediately flushed in with water.

CONCLUSION

Plants can carry out both photosynthesis and respiration simultaneously. During

photosynthesis, plants are using the energy of the sun to build molecules which effectively store

this energy (glucose). Chemically, the photosynthetic reaction looks like this:

6CO2 + 6H20 C6H12O6 + 6O2

During respiration, plants are using this stored energy (glucose), to fuel their metabolic processes.

Chemically, the respiratory process looks like this:

C6H12O6 + 6O2 → 6CO2 + 6H20 + energy

On the other hand, the converted energy from respiration is used to synthesize molecules,

move materials around within the organism, grow (create new cells) and reproduce. Notice that in

photosynthesis, CO2 (carbon dioxide) is being used up as it is fixed into glucose molecules. During

respiration the opposite is true. As the plant releases the energy stored in glucose by breaking it

down, CO2 is being given off into the surrounding water or atmosphere. The relationship between

these two processes is special in that it allows plants to recycle some of their by-products. (While

CO2 is being given off during respiration, it can be re-utilized during photosynthesis).

In this experiment, we are demonstrating the net change in carbon dioxide when the

common fresh water plant Elodea is placed under different conditions. We used a pH indicator,

bromothymol blue, as a means of determining the presence or absence of CO 2. A solution of

bromothymol blue changes color when CO2 is introduced. It changes color due to a change in pH.

When CO2 is dissolved in water, it forms carbonic acid. This lowers the pH of the solution and

causes the bromothymol blue to change its appearance.