EX: NO:1 POLLEN GERMINATION

Aim
The experiment aims to observe pollen germination on a slide.
Materials Required
Freshly plucked flower from grass or china rose,

10 mg boric acid
Beaker
Microscope
Sucrose (10 g)
Coverslips

Slide
Dropper
Magnesium sulphate (20 mg)
Calcium nitrate (30 mg
Procedure

The first step is to prepare the nutrient solution by dissolving the sucrose and boric acid in
water. Then this mixture has to be dropped using a dropped on the cavity slip. Then using
a brush or simply by fingers, brush off a few pollen grains and let the slide for 15 minutes.
Then the microscope has to be used to observe the slide at intervals of 30 minutes.
Observations
It can be observed that by enlargement of the vegetative cells, the pollen grains are
germinating when submerged in the nutrient medium. A pollen tube is formed when the
nucleus grows and emerges from one of the germ pores and forms two male gametes which
can be either spherical or lenticular in shape. These pollen grains that germinate are called
viable pollen grains. These gametes are called sperm nuclei. There will also be some pollen
grains that will not form pollen tubes, hence, they are called non-viable pollen grains.

Precautions
There are precautions to be observed while performing this experiment:
Dirty or previously used slides should not be used for dusting pollen grains. Slides
should be clean and dry for use.

Only a few drops of the nutrient medium is required for germination. 2-3 drops of
the solution are enough, more than that may hinder the process of pollen
germination.
 Only freshly plucked flowers should be used and the experiment should be
conducted within some time of plucking.

The cavity slide used for observation should have depression at the centre so that
the solution can be held in that cavity and does not flow.
Conclusion
Pollen germination can be carried out in a nutrient medium and studied under a
microscope when proper environmental conditions are given.
DIAGRAM :

EX : NO:2 PLANT POPULATION DENSITY BY THE QUADRANT METHOD.

Aim
To study the plant population density by the quadrant method.

Materials Required

• Nail.
• Thread
• Hammer

Procedure

1. Select a site for the study and hammer the nails on the site without harming the
vegetation.
2. Fix four nails in the form of a square.
3. Each end of the nail is tied with the help of a thread making a 1m*1m quadrant.
4. Nine more similar quadrants are made at the site of the study.
5. The number of individuals of species A present in the first quadrant is counted and
the data is recorded in the table.
6. The number of individuals of species A in other quadrants is also counted and the
data is recorded in the table.
 7. Similarly, count the number of individuals of species B and C present in all the
quadrants and record the data in the table.
8. The density of the plant population is then calculated by the following equation:

Conclusion
The population density is the highest for species A and the lowest for species C. The density
value is expressed as the number of individuals per unit area.

EX: NO: 3 PLANT POPULATION FREQUENCY BY THE QUADRANT METHOD.

Aim
To study the plant population frequency by the quadrant method.

Materials Required

• Cotton/Nylon thread
• 4 nails
• Hammer

Procedure

1. Select the site of study and make a quadrant of 1m*1m using the nails and the thread.
2. Fix the nails with the help of a hammer without destroying the vegetation.
3. Make nine similar quadrants at the site of study.
4. The plant species for the study should be selected.
5. Observe the species in the first quadrant and mark them as species A.
6. Check the presence of species A in all the quadrants and record the observations in the
table.
7. Similarly, record the number of species B and C in all the quadrants and mention them in
the table.
8. Determine the frequency of plant population by the formula:

Conclusion
The plant population frequency is the highest in species C and the least in species A. It shows
how many times a plant species is present in the provided number of sample quadrats.
PreparPrepare A Temporary Mount of the Onion Root

EX: 04 PREPARING A TEMPORARY MOUNT OF AN ONION ROOT TIP

TO STUDY MITOSIS
Aim
To study mitosis by preparing a temporary mount of an onion root tip.

Necessary Materials & Apparatus

 • Onion
• Watch glass
• Glass slide
• Filter paper
• Aceto-alcohol
• Coverslip
• Water
• N/10 Hydrochloric acid
• Acetocarmine Stain
• Burner
• Forceps
• Dropper
• Blade
• Needle
• Compound microscope

Procedure

• Place the onion on a tile

• Using the blade, remove the dry roots
• Regrow the root tips by placing the bulbs in a water-filled beaker
• After 3 to 6 days, new roots may emerge
• Slide 2 to 3 cm off freshly grown roots and place them on a watch glass
• Use a forceps to transfer the freshly cut tips to a test tube containing Aceto-alcohol (1:3 =
anhydrous acetic acid: ethanol)
• Submerge the root tips in the solution for 24 hours
• Use the forceps to take out a single root and place it on a glass slide.
• Put a single drop of N/10 HCl on the root tip
• Then, put 2-3 drops of acetocarmine stain
• Use a burner to warm it, and ensure that the stain does not dry up.
• Use a filter paper to blot out the excess stain, if any
• Cut the significantly stained portion of the root using a blade and place it on a slide.
Discard the rest of the root
• Put a drop of water on the root tip
• Place a coverslip using a needle
• Tap the coverslip such that the meristematic tissue of the root tip is compressed and
spread out as a thin layer.
• The preparation is ready for studying mitosis.

Observation
Use the compound microscope to study the various phases of mitosis.
Exercise 5: Isolation of DNA from plant materials
DNA is one of the nucleic acids found in living systems. DNA acts as the genetic
material in most of the organisms.

Aim:

To isolate DNA from available plant materials such as spinach leaves, fresh green
pea seeds, green papaya, etc.

Requirements:

Plant materials, mortar and pestle, beakers, test tubes, ethanol, etc.

Principle:

Recombinant DNA technology has allowed breeders to introduce foreign DNA in

other organisms including bacteria, yeast, plants and animals. Such organisms are
called Genetically Modified Organisms (GMOs). Thus rDNA technology involves
isolation of DNA from a variety of sources and formation of new combination of
DNA.

Procedure:

Take a small amount of plant material and grind it in a mortar with a little amount
of water and sodium chloride.Make it into a solution and filter it. To this filterate,
add liquid soap solution or any detergent solution and mix it with a glassrod. Then
tilt the test tube and add chilled ethanol and leave it aside in the stand. After half-
an-hour we can observe the precipitated DNA as fine threads. DNA that separates
can be removed by spooling DNA that separates can be removed by spooling

Observation:

DNA appears as white precipitate of very fine threads on the spool.

Inference:

Thus DNA can be isolated from the plant cell nucleus by this technique.

Precautions:

• All the glasswares must be thoroughly cleaned and dried.

• The chemicals used for the experiments must be of standard quality.

• If ordinary ethanol is used, the time duration for obtaining precipitated DNA
may extend further.
 SPOTTERS 1

Adaptations of flowers for pollination by different agents.

Aim: To study the adaptations in flowers for pollination by different agents (wind
and insects)

Principle: The process of transfer of pollen grains from the anther to stigma of a
flower is called pollination.

Requirements: Fresh flowers of maize or any other cereal / gram, any insect
pollination flowers like Salvia, Calotropis, Ocimum and Asteraceae flowers.

Place the given flower on a slide and observe it with the help of hand lens. Note
down the adaptations of the flowers meant for pollination by the external agents.

A. Wind Pollinated Flowers – Anemophily

Diagnostic Features

• The flowers are small, inconspicuous, colourless, odourless and nectarless.

• Anthers and stigmas are commonly exerted.

• Pollen grains are light, small, powdery and produced in large numbers.

• The stigmas are large, sometimes feathery and branched adapted to catch the
pollens.
 B. Insect Pollinated Flowers – Entemophily

Diagnostic Features

• The flowers are showy, brightly coloured and scented.

• The flowers produce nectar or edible pollen.

• Anthers and stigmas are commonly inserted.

• Stigmas are usually unbranched and flat or lobed.

SPOTTERS 2
POLLEN GERMINATION ON STIGMA THROUGH A PERMANENT SLIDE.

Aim
To observe pollen germination on stigma through a permanent slide.

Materials Required
1. Microscope
2. Permanent slide

Procedure
The permanent slide is placed under a microscope and the pollen germination is observed.

Theory
Pollination refers to the transfer of pollen grains from the anther of a flower to the stigma of
the same or different flower through biotic or abiotic means. The complete process of
pollination is as listed below:

• Once the pollen grains are deposited on the stigma, it starts to germinate with the
absorption of nutrients and water.
• A small pollen tube is produced through the style to the ovary.
• The tube cell moves out of the pollen grain and through one of the germ pores forms
a pollen tube.
• The nucleus of the tube moves down to the tip of the pollen tube.
• The generative cells also pass into it and soon divide to form two male gametes.
• During double fertilization, one of the two sperms fuses with the egg cell of the ovule.
This helps in embryo development.
• The other cell combines with another subsidiary nuclei of the ovule that helps in the
formation of the endosperm.
• The growing ovule is transformed into a seed.

SPOTTERS 3

Identification of Stages of Gamete Development, i.e., T.S. of Testes and T.S. of Ovary
Through Permanent Slides (from Grasshopper, Mice)

Aim
To identify the stages of gamete development, i.e., T.S. of testes and ovary through permanent slides
from mice.

Materials Required
• Permanent slides of T.S. of testes.
• Permanent slides of T.S. of the ovary.
Procedure

T.S. of Testes of Mice

• The testes comprise several seminiferous tubules embedded in the interstitial tissues.
• Thick fibrous tissues called tunica albuginea cover the testes.
• It comprises different types of cells from the outside to the lunar in the manner given below:
Spermatogonia → Spermatocytes → Spermatids → Spermatozoa (sperms)

• Sertoli cells are located between the germinal cells.

• The Leydig cells that produce testosterone are present in the interstitial tissues.

T.S. of Ovary of Mice

• An ovary is a germinal epithelium bounded by a solid structure covered by a thick layer of
fibrous tissue known as tunica albuginea.
• It consists of an inner medulla and an outer cortex.
• The medulla comprises several round or oval bodies known as ovarian follicles.
• Follicle development takes place in the following stages:
1°follicle → 2°follicle → 3°follicle → Graffian follicle → Corpus luteum

• Cortex comprises corpus luteum along with mature follicles.

Precautions
• The microscope should be handled with care.
• Adjust the lens such that the focus is better.