Experiment – 1 - Preparing a temporary mount of a leaf peel to show stomata.

Aim
To prepare a temporary mount of a leaf peel in order to show the stomata of a leaf

Principle/Theory
Stomata are tiny openings or pores which are commonly noticed being distributed on the
epidermis of the leaves and also in young stems. Typically, stomata are found on the lower
surface of the leaves. They are surrounded by two-kidney shaped cells known as guard cells
on either side of the stomata. The guard cells possess a thick inner wall and a thin outer
covering which control the closing and opening of the pores of stomata. Stomata carries out
the function of regulating gas exchange and water vapor between the leaves of the plant and
the atmosphere. Turgidity of the guard cells causes the stomata to open while the flaccid
nature of the guard cells causes the stomata to close.

Materials Required

 Tradescantia leaf (Rheo leaf)

 Needles
 Forceps
 Watch glass
 Dropper
 Glass slides
 A brush
 Coverslips
 Blotting paper
 Safranin
 Compound microscope
 Glycerin

Procedure

 Pick a healthy leaf from the potted plant

 Fold the leaf to gently pull the peel apart to separate a peeled section from the lower
surface of the leaf. Use the forceps to perform this step. Allow the peel to remain in a
watch glass holding water for some time.
 In the watch glass, stain the sample by adding some drops of safranin through a
dropper.
 Take the peel out after 2-3 minutes. Set it on a clear glass slide
 Add a drop of glycerin to the peel. Put a clear coverslip over it gently using a needle.
 Excess glycerin and stain can be removed using blotting paper
 Examine the slide first under a low-power and then under a high-power magnification
of a compound microscope.

Observation

 Visible epidermal cells. The cells in their outline are irregular with no intercellular
spaces

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  Small openings, stomata are scattered through the epidermal cells
 Guard cells are observed which have chloroplasts and nucleus
 Guard cells are observed having a thin outer covering and a thick inner
boundary(concave)
 Guard cells control the closing and opening of the stomata.

Figure - Temporary mount of a leaf peel - Opening and closure of stomata

Conclusion
Epidermal cells are found containing stomata on the lower surfaces of the leaf.

Precautions

 Avoid folding the leaf too much. The peel should be snipped to a proper size
 The peel should always be placed at the Centre of the slide and the slides should be
held from the sides.
 The peel should neither be over-stained nor under-stained
 A brush should be used to handle the peel, otherwise it would damage cells.
 Glycerin should be used in order to prevent drying of the peel
 Coverslip needs to be placed in such a way that air bubbles are avoided
 Blotting paper can be used to remove excess stain

Experiment 2 - Experimentally show that carbon dioxide is given out during

respiration.

Aim
To experimentally demonstrate that carbon dioxide is released during the process of
respiration.

Principle/Theory
The process of respiration is biochemically carried out wherein food, glucose to be precise,
is oxidized and energy is released. In this experiment, gram seeds (moistened) are used.
The purpose of using these seeds is that they release carbon dioxide and are respiring
actively. The released carbon dioxide is consumed by the solution of KOH.

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Materials Required

 Soaked gram seeds

 U-shaped delivery tube
 Conical flask
 Blotting paper (moist) /cotton wool
 Thread
 Water
 Beaker
 Test tube
 Rubber cork with a single hole
 Freshly prepared KOH solution (20%)
 Vaseline

Procedure

 Germinate close to 25 seeds. This can be done by wrapping them in moist blotting
paper or cotton wool for around 3 to 4 days.
 Set up the germinated or sprouted seeds in the conical flask. Spray some water into
the flask to dampen the seeds.
 With the help of a thread, suspend the conical flask containing the test tube having a
freshly prepared 20% KOH solution.
 Use the rubber cork to seal the opening of the conical flask.
 One edge of the U-shaped glass delivery tube present in the conical flask should be
inserted through the hole in the rubber cork. The other edge should be placed into a
beaker that is saturated with water.
 All attachments to the set-up should be sealed. This can be done using Vaseline to
create an air-tight environment.
 The initial water level present in the U-shaped delivery tube needs to be marked.
 Leave the experimental set-up uninterrupted for 1 to 2 hours. Observe the
fluctuations in the water level in the tube.

Observation
Careful observation after a certain period of time reveals that the water level in the U-shaped
delivery tube has risen in the beaker.

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Conclusions
The rise in water level indicates that carbon dioxide is released as a result of germinating
gram seeds during the process of respiration in the conical flask. The carbon dioxide that is
released in the process is absorbed or consumed by the KOH solution that is suspended in
the test tube in the conical flask, creating a vacuum or a void in the flask resulting in the
upward water movement in the tube. Hence, the water level in the tube changes.

Precautions

 The seeds that are to be germinated need to be moistened

 Air-tight environment for all the connections in the experimental set-up
 The KOH solution that is used needs to be freshly prepared
 Care needs to be taken to ensure that one end of the delivery tube is placed in the
conical flask. The other edge is submerged in the water of the beaker
 The tube that contains the KOH solution needs to be suspended carefully

EXPERIMENT - 3: Studying (a) binary fission in Amoeba, and (b) budding in yeast and
Hydra with the help of prepared slides

Aim
To study about (a) Binary Fission in amoeba and (b) Budding in yeast with the help of
prepared slides

Materials Required

 Compound microscope
 Permanent slides of budding in yeast and binary fission in amoeba

Procedure

 Place the slide under a compound microscope

 Focus the slide, first under low power and later under high power of the compound
microscope
 Various stages of budding and binary fission can be carefully examined

Observation
(a) Binary fission in Amoeba

 Initially, the pseudopodia are retrieved. The body of amoeba is coiled and becomes
round
 Amitosis is observed, the division of the nucleus takes places which are followed by
splitting of cytoplasm
 At the point of fission in the body of the amoeba, a construction starts to develop.
 The constriction or furrow turns deeper resulting in the formation of two daughter
cells

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(b) Budding in yeast

 Protuberance or a tiny outgrowth is observed on the parent cell

 Division of the nucleus is observed which is later seen in the bud
 Repetitive budding leads to the formation of a chain of cells

Figure – Budding in Yeast

(c) Budding in Hydra

 Hydra uses its regenerative cells for forming buds

 Bud develops as an outgrowth due to repeated cell division at one specific site
 Buds detach from parent body when fully matured and develop into new hydra

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Conclusions
The prepared slides display asexual reproduction. One individual is involved in producing a
new offspring of their own kind.

Precautions

 Slides need to be aligned and focused accurately

 Sketch out your observation that is observed under a microscope
 The slides first need to be examined under a low-power magnification of the
compound microscope and then under high-power magnification.

EXPERIMENT -4: Identification of the different parts of an embryo of a dicot seed (Pea,
gram or red kidney bean)
Male and female gametes fuse to form the zygote during sexual reproduction. The zygote
further undergoes division to evolve into an embryo. Monocots and dictators do not differ in
the initial stages of development, but they differ in the later stages of development.

Aim
To identify the different parts of an embryo of a dicot seed

Principle/Theory
The process of fertilization in plants leads to the formation of fruits which form the ripened
ovary. The seed can be one or many which form the mature ovule.
An embryo consists of the following parts: radical (from which roots arise), plumule (from
which shoots arise) and cotyledons (nourishes the seeds)

Materials Required

 Seeds of red kidney bean/gram

 Forceps
 Magnifying glass
 Cloth
 Petri dish
 Water

Procedure

 Soak a few seeds overnight

 Next morning, drain the excess water out
 Now wrap the seeds in a clean and a moist cloth for a day, allow it to dry
 Next, carefully peel the seed coat
 With the help of forceps, dissect the seed so as to get two equal halves
 Examine with the help of a magnifying glass. Carefully identify and locate different
parts of the seed
 Sketch out the interior of the seed you examined labeling all the parts as shown in
the diagram.

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 Figure- Embryo of a dicot seed

Observation

 The bean seed resembles the shape of a kidney. It has a convex and a concave side
 A scar known as the hilum is observed on the slightly darker side of the concave side
 A tiny pore known as the micropyle is located just adjacent to the hilum
 The seed is enclosed by a seed coat
 The embryo possesses two distinct and large cotyledons that resemble the shape of
a kidney and are white in color
 Lateral attachment of the cotyledons to the curved embryonal axis is observed
 Radicle is examined. It is the rod-shaped and lightly protrusive lower end of the
embryonal axis that is found placed towards the micropylar end.
 The upper end of the embryonal axis exhibits the plumule
 Hypocotyl is observed which is a section of the embryo axis found in between the
radicle and adjunct of cotyledon leaves
 The epicotyl is also observed which is the section of the embryo axis between the
adjunct of cotyledon leaves and plumule

Conclusion
Three principal parts of the embryo of dicot seeds are observed, they are:

 Cotyledons
 Plumule
 Radicle

Precautions

 Care needs to be taken while dissecting the seed as it may damage the seed
 The cloth that is used to wrap the seeds needs to be moist