Class XII

Biology
Instructions
1. Do not write Dates
2. Make all diagrams on blank side
3. Make diagrams with pencil no coloring and shading is allowed
4. Make observation table on left side/blank side with pencil
5. Fill the index mention experiment detail and page number and dated (when told)

Experiment No 1

Aim: To calculate percentage of pollen germination

Principle: In nature, pollen grains germinate on the compatible stigmas of the carpel. Pollen grains
can also be induced to germinate in a synthetic medium. During germination, the intine (inner wall) of
pollen grain emerges as a pollen tube through one of the germ pores in the exine (outer wall).

Requirement: Calcium nitrate, boric acid, sucrose, distilled water, petridish, slides, coverslips,
brush, needle, microscope, and mature pollen grains of Tradescantia/balsam/
Jasmine/lily/pomegranate/grass/Vinca/China rose/Petunia.

Diagram on plain side

Procedure
(i) Prepare the pollen germination medium by dissolving 10g sucrose, 30mg calcium nitrate and 10mg
boric acid in 100ml of distilled water. Alternatively 10% sucrose solution can also be used. (ii) Take a
drop of medium or 10% sucrose solution on a cover slip and sprinkle mature pollen grains on the
drop.
(iii) Invert the cover glass on to a slide
(iv) After 10 minutes, observe the slide under microscope

Observation

Several pollen grains germinate and put forth pollen tubes.

Precautions
1.Only a few drops of the nutrient medium is required for germination. ...
2.Only freshly plucked flowers should be used and the experiment should be conducted within some
time of plucking.

Experiment No 2

Aim
To study the flowers adapted to pollination by different agents such as wind, insects and birds.

Material Required

● Fresh flowers
● Magnifying glass

Theory

Pollination is the process of transferring pollen from the male anther of a flower to the female

stigma of the same or different flower. Pollination can be carried out by different agents such as

wind, water, birds, insects, etc.

Following are a few observations of the flowers that are adapted to pollination by wind, insects and

birds.
Flowers Pollinated By Wind

Most of the conifers and angiosperms exhibit wind pollination. Such flowers do not produce nectar

and fragrance. In the flowers pollinated by the wind, the microsporangia hang out of the flower. As

the wind blows, the light-weight pollen blows with it. The pollen gets accumulated on the feathery

stigma of the flower. These flowers appear even before the leaves when the spring commences. Few

examples of such flowers include:

● Rice
● Corn
● Oats
● Maize
● Barley
● Papaya

Flowers Pollinated By Insects

The flowers pollinated by insects are bright-coloured and produce nectar. The fragrance of the

flowers attracts the insects. The pollen is sticky, large, heavy and rough so that it sticks to the

body of the insects. The stigmas are also sticky so that the pollen deposits are not dispersed.

Nectar guides are present on the petals. Few examples of the flowers pollinated by insects are:
 ● Aster
● Lithops
● Magnolia

Flowers Pollinated By Birds

The flowers pollinated by birds are strong and are adapted to allow the birds to stay near the

flowers without their wings getting entangled in them. The flowers are tubular and curved that

facilitates nectar-sucking by birds. The flowers are odourless and bright-coloured that attracts

the birds. While sucking the nectar, the pollen gets deposited on their beaks and neck and is

transferred to the plant they visit next. Few examples of flowers pollinated by birds include:

● Hibiscus
● Fuchsias

Experiment No 3

Aim

To observe pollen germination on stigma through a 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.
● Pollen grains are seen adhered to the stigma surface. Depending on the plant species, the
pollen grains may vary in shape (spherical, oval, or spiky) and size.
● The stigma may appear as a slightly raised, sticky, or papillate surface to which pollen grains
adhere.

Observation
 ● Pollen grains are seen adhered to the stigma surface.
● In some pollen grains, emergence of the pollen tube from the germinal pore of the grain is
observed.
● The pollen tubes appear as fine, tubular extensions that are often slightly transparent and
can vary in length depending on the stage of germination.
● The pollen tubes grow downwards into the stigma, directed towards the ovary. The direction
of growth is often visible in a successful pollen grain.

Experiment No 4

Controlled pollination is a crucial technique in plant breeding used to produce desired hybrids by
preventing unwanted pollination and ensuring cross-pollination between selected parent plants. This
process ensures genetic control over the offspring by manipulating the pollination environment.
Controlled pollination often involves emasculation (removal of male reproductive organs), tagging
(labeling the flowers), and bagging (covering the flowers to prevent contamination by other pollen
sources).

Emasculation:

Emasculation is the removal of male reproductive organs (anthers) from a flower before the pollen
is released. This prevents self-pollination and allows controlled cross-pollination from a desired male
parent.
 ● Emasculation must be performed before the anthers mature and release pollen. This
typically happens in the bud stage.
● Fine forceps or scissors are used to carefully remove the anthers without damaging the
female reproductive parts (stigma and style).
● Emasculation ensures that the female flower will only receive pollen from the selected male
parent, thus preventing self-pollination or accidental pollination.
● Emasculation is widely practiced in plants like wheat, maize, rice, and many horticultural
crops where cross-pollination is needed to create hybrids.
● Once emasculation is complete, the flowers are protected to avoid unwanted pollen
exposure, typically through bagging.

Bagging:

Bagging is the practice of covering flowers or inflorescences with a protective bag after
emasculation and pollination to prevent contamination from unwanted pollen.

● After controlled pollination, bagging ensures that no external pollen can reach the stigma,
thereby maintaining the integrity of the cross-pollination process.
● Bags are typically made of paper, cloth, or plastic, depending on the crop and environmental
conditions. The material should allow air circulation while preventing pollen entry.
● In some cases, bagging is done before the flowers open to prevent any exposure to
unwanted pollen even before emasculation. This is called pre-bagging.
● After pollination, the flowers are bagged again to ensure only the desired pollen fertilizes
the ovules, and no other pollen grains contaminate the cross.
● Bagging must be monitored regularly to ensure the flowers are not damaged, and the bags
are securely in place until fertilization is complete.

Tagging:

Tagging refers to the practice of labeling flowers that have undergone controlled pollination. It
helps track and identify the crosses made during breeding programs.

Tags usually contain information like the date of emasculation, pollination, the male and female
parent plant names or codes, and other relevant details for tracking

Experiment No 5

Aim: To study the discrete stages of gametogenesis in mammalian testis and ovary
Principle: In all male and female organisms gamete formation takes place in their gonads, i.e., testis
and ovary respectively. The process of gamete formation, called gametogenesis involves meiotic cell
division. The gametogenic development in testis is called spermatogenesis and in ovary it is
oogenesis. They exhibit marked differences and can be examined in transverse section (T.S.) of
these organs.
TS of Testis

Observation
Seminiferous Tubules: Circular or oval structures where spermatogenesis occurs; lined with
germinal epithelium containing developing sperm cells.
Germ Cells: Different stages of sperm development observed, from spermatogonia near the
basement membrane to mature spermatozoa in the lumen.
Sertoli Cells: Large, columnar cells providing support to germ cells; their elongated nuclei are
prominent within the seminiferous tubules.
Leydig Cells: Found between seminiferous tubules, these cells secrete testosterone and appear in
clusters with large, round nuclei.
Tunica Albuginea: Thick, fibrous outer layer providing structural support to the testis, visible
around the edge of the section.

T.S. of Ovary
(i) In the section of ovary, there is a mass of tissue lined with germinal epithelium. Inside that you
will observe an ovum, which is a cell surrounded by one to several layers of follicular cells. As the
ovum matures, the number of surrounding follicular cell layer increases ..
(ii) In the later stage of follicular development a cavity called antrum appears.
(iii) The cavity gets further enlarged and the follicle grows bigger. This is the stage of Graafian
follicle ready to release the ovum (ovulation).
(iv) In the next stage, you may notice a Corpus luteum, and/or Corpus albicans, which differ from
each other and also from Graafian follicle in their features.

Experiment No 6

Aim: To study the blastula stage of embryonic development in mammals, with the help of permanent
slide, chart, model or photograph
Principle: The zygote undergoes a few cycles of mitotic divisions to form a solid ball of cells called
morula. The cells continue to divide and at a later stage a cavity is formed within it. This stage is
blastula. The internal structural details of blastula can be observed in its transverse section.
Observation
In a transverse section, the blastula appears as a hollow sphere with a central cavity known as the
blastocoel. The outer layer of cells surrounding the blastocoel is called the trophoblast. These cells
will eventually contribute to the formation of the placenta. On one side of the blastula, attached to
the trophoblast, is a group of cells called the inner cell mass (ICM), which will give rise to the
embryo itself. This arrangement is key in early embryonic development, as the inner cell mass
differentiates into the different tissues and organs of the organism.

Experiment no 7

Aim

To observe the stages of meiosis in onion bud cell or grasshopper testis through permanent slides.

Principle: Meiosis is a type of cell division in which the number of chromosomes is halved (from

diploid to haploid) in the daughter cells, i.e., the gametes. The division is completed in two phases,

meiosis I and meiosis II. Meiosis I is a reductional division in which the chromosomes of homologous

pairs separate from each other. Meiosis II is equational division resulting in the formation of four

daughter cells. Stages of meiosis can be observed in a cytological preparation of the cells of testis

tubules or in the pollen mother cells of the anthers of flower buds.

Observations

The different stages of meiosis are observed along on the basis of the following features.
Stages of Meiosis I

Prophase I

In this stage, the chromosomes condense and move towards the centre of the cell. It consists of

five different sub-phases:

● Leptotene: The homologous chromosomes replicate.

● Zygotene: Synapsis between homologous chromosomes start.
● Pachytene: The sister chromatids separate but the homologous chromosomes remain
attached.
● Diplotene: The two homologous chromosomes migrate apart and disintegrate between the
chromosomal arms.
● Diakinesis: The condensation of chromosomes stops at this stage and the chiasmata is
clearly visible under an electron microscope. The nucleolus and the nuclear envelop disappear
at this stage and the centrosome moves to the equator.

Metaphase I

The homologous chromosomes that contain two different alleles for each gene, line up on the

metaphase plate to be separated.

Anaphase I

The separated chromosomes are pulled towards the centrioles on either side of the cell.

Telophase I

The chromosomes are completely pulled apart and new nuclear envelope forms.

Stages of Meiosis II

Prophase II

In this stage, the nuclear envelope disintegrates and centrioles develop.

Metaphase II
The chromosomes line up on the metaphase plate and the chromatids are on either side of the

metaphase plate.

Anaphase II

The sister chromatids separate and are known as sister chromosomes.Telophase II

The cell divides into two and a new nuclear envelope surrounds the chromosomes.

Experiment No 8

Aim: Preparation and study of mitosis in onion root tips

Principle: Somatic growth in plants and animals takes place by the increase in the number of cells. A
cell divides mitotically to form two daughter cells wherein the number of chromosomes remains the
same (i.e., unchanged) as in the mother cell. In plants, such divisions rapidly take place in
meristematic tissues of root and shoot apices, where the stages of mitosis can be easily observed.
In animals, mitotically dividing cells can be easily viewed in the bone marrow tissue of a vertebrate,
epithelial cells from gills in fishes and the tail of growing tadpole larvae of frog.

Requirement: Onion bulbs, wide mouth glass tubes, glacial acetic acid, ethanol 2-4% acetocarmine
stain, N/10 HCl, spirit lamp, slide, cover slips, blotting paper, and compound microscope.

Procedure:
1. Take one or two preserved roots, wash them in water on a clean and grease free slide.
2. Place one drop of N/10 HCl on the root tip followed by 2–3 drops of aceto-carmine or
aceto-orcein stain on it. Leave the slide for 5–10 minutes
3. Carefully blot the excess stain using blotting paper. Now cut the comparatively more stained
(2–3 mm) tip portion of the root and retain it on the slide and discard the remaining portion.
4. After (10–20 seconds) put one or two drops of water and blot them carefully using blotting
paper. Again put a drop of water on the root tip and mount a cover slip on it avoiding air
bubbles.
5. Place the slide in between the folds of blotting paper using the fingers in such a way that
the cover slip mounted on the slide is properly held.
6. Now slowly tap the cover slip using the blunt end of a pencil so that the meristematic tissue
of the root tip below the cover slip is properly squashed and spread as a thin layer of cells.
7. Place the slide on the stage of a good quality compound microscope. First observe it under
the lower magnification (10 X objective) to search for the area having a few dividing cells.
 Examine the dividing cells under higher magnification of the microscope to observe the
detailed features of mitosis.

Observation:

Make the diagrams on blank side

The stages of mitosis can be broadly categorised into two parts: karyokinesis (division of nucleus)
followed by cytokinesis (division of cytoplasm, and ultimately of the cell). Those cells, which are not
in the phases of cell division are considered to be in interphase.
Interphase
The cells are mostly rectangular, oval or even circular in shape, with almost centrally situated
densely stained nucleus. The chromatic (coloured) material of the nucleus is homogeneous and looks
granular. The boundary of the nucleus is distinct. One or few nucleoli (sing: nucleolus) can also be
observed inside the nucleus.

Stages of Mitosis
(a) Prophase
Intact nuclear outline is seen. The chromatin (seen as a homogeneous material in the nucleus at
interphase) appears as a network of fine threads (chromosomes). Nucleoli may or may not be visible
if the cell under observation is in the early stage of prophase then the chromatin fibres
(chromosomes) are very thin. However, in the cells at late prophase, comparatively thicker
chromatin fibres would be visible. Besides this, in the late prophase the nuclear membrane may not
be noticed.
(b) Metaphase
The nuclear membrane disappears. Chromosomes are thick and are seen arranged at the equatorial
plane of the cell. Each chromosome at this stage has two chromatids joined together at the
centromere, which can be seen by changing the resolution of the microscope. Nucleolus is not
observed during metaphase.
(c) Anaphase
This stage shows the separation of the chromatids of each chromosome. The chromatids separate
due to the splitting of the centromere. Each chromatid now represents a separate chromosome as it
has its own centromere. The chromosomes are found as if they have moved towards the two poles
of the cell. The chromosomes at this stage may look like the shape of alphabets 'V', 'J' or 'I'
depending upon the position of centromere in them. Different anaphase cells show different stages
of movement of chromosomes to opposite poles, and they are designated to represent early, mid
and late anaphase.
(d) Telophase
Chromosomes reach the opposite poles, lose their individuality, and look like a mass of chromatin.
Nuclear membrane appears to form the nuclei of the two future daughter cells.
Cytokinesis
In plants, a cell plate is formed in the middle after telophase. The plate can be seen to extend
outwards to ultimately reach the margin of the cell and divide the cell into two. Such cell plates are
characteristic of plant cells . However, in an animal cell, the two sides of the cell show inpushings or
constrictions formed from the peripheral region in the middle of the cell, which grow inward and
meet to divide the cell into two daughter cells.

Precautions
1. Use freshly grown onion roots, as dried or old roots are not suitable for the experiment.
2. Place the onion bulb in a glass jar filled with water and make sure the base of the bulb
touches the water. Keep the jar in a safe place for 3–6 days, or until the roots have grown
2–3 cm.
3. Cut off 1–3 cm of the freshly grown roots

Experiment No 9
Mendelian inheritance using seeds of different colour/sizes of any plant.
Experiment no 10

Prepared pedigree charts of any one of the genetic traits such as rolling of tongue, blood groups,
ear lobes, widow's peak and colour blindness.
Experiment No 11

Flash cards models showing examples of homologous and analogous organs

Experiment No 12
Common disease causing organisms like Ascaris, Entamoeba, Plasmodium, any fungus causing
ringworm through permanent slides, models or virtual images or specimens. Comment on symptoms
of diseases that they cause.
Experiment No 13

Isolate DNA from available plant material such as spinach, green pea seeds, papaya, etc.
Experiment No 14
Models specimen showing symbolic association in root nodules of leguminous plants, Cuscuta on host,
lichens.

Experiment No 15
Study the plant population density by quadrat method.
Experiment No 16

Study the plant population frequency by quadrat method