EXPERIMENT NO.

: 1

Aim: To study pollen germination on a slide.

Principle:

1. In nature, pollen grains germinate on the compatible stigmas of the carpel. Pollen grains can also be induced to
germinate in a synthetic medium.
2. During germination The pollen grains will germinate when submerged in the sugar rich nutrient medium. This is
characterized by the enlargement of the vegetative/tube cell.
3. It emerges through one of the germ pores to form a pollen tube. The generative cell nucleus makes two male
gametes (sperm nuclei).

Requirements:
Freshly plucked seasonal flowers Petunia, beaker, boric acid, sucrose, microscope and cavity slide.

Procedure:

1. The first step involves the preparation of a sugar solution. This is done by dissolving 10g of sucrose 90ml of
water.
2. Pour a few drops of this solution onto the cavity slide.
3. use a brush or fingers to gently dust a few pollen grains from the stamen of mature flowers.
4. Let the slide set for 5 minutes. Then, use the microscope to view the slides in 30- minute intervals.

Observation:

Different stages of germinating pollens are observed. Some pollens are in their initial stage of germination while others
have quite long pollen tube containing tube nucleus and two male gametes.

DIAGRAM TO BE DRAWN ON LEFT SIDE OF OBSERVATION (PLAIN PAPER WITH PENCIL)

Precautions:

1. Flowers should be freshly plucked.

2. Use clean cavity slide to observe the pollen grains.
3. The slides should not be disturbed, otherwise position of pollen grains will get changed.
4. During observations pollen grains must be properly dipped in nutrient solution.
 EXPERIMENT NO. : 2

AIM: to study the Flowers adapted to pollination by different agencies (wind, insects, and
birds).

Flowers adapted to pollination by BIRDS

COMMENTS:
1. Pollination is the process of transferring pollen from the male anther of a flower
to the female stigma of the same or different flower.
2. Pollination of flowers by insects is called ornithophily.
3. 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.
4. The flowers are tubular and curved that facilitates nectar-sucking by birds.
5. 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.
6. Few examples of flowers pollinated by birds include: Hibiscus, Bignonia,
Verbenas

Flowers adapted to pollination by WIND

COMMENTS:
1.Pollination is the process of transferring pollen from the male anther of a flower to the female stigma of the same or
different flower
2. Most of the conifers and angiosperms exhibit wind pollination. Pollination of flowers by the wind is called as
anemophily.
3.Such flowers do not produce nectar and fragrance.
4. 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.
5.These flowers appear even before the leaves when the spring commences.
6.Few examples of such flowers include: Rice, Barley, Papaya, Maize,Oats

7.
Flowers adapted to pollination by INSECTS
COMMENTS:
1. Pollination is the process of transferring pollen from the male anther of a flower to the female stigma of the same
or different flower.
2. Pollination of flowers by insects is called entomophily.
3. The flowers pollinated by insects are bright-coloured and produce nectar. Nectar guides are present on the petals.
4. The fragrance of the flowers attracts the insects.
5. The pollen are sticky, large, heavy and rough so that stick to the body of the insects.
6. The stigmas are also sticky so that the pollens depositing are not dispersed.
7. Few examples of the flowers pollinated by insects are: Salvia, Datura, Gulmohar,Calatropis etc

EXPERMINT NO. :4

AIM: Identification of stages of gamete development, i.e., T.S. of testis and T.S. of ovary through
permanent slides (from grasshopper/mice).

Requirements: permanent slides of T.S. of testis and L.S. of ovary, Microscope

COMMENTS:
 T.S. OF TESTIS
1. The testes comprise several seminiferous tubules embedded in the interstitial tissues.
2. Thick fibrous tissues called tunica albuginea cover the testes.
3. It comprises different types of cells from the outside to the lunar in the manner given below:
Spermatogonia → Spermatocytes → Spermatids → Spermatozoa (sperms)
4. Sertoli cells are located between the germinal cells.
5. The Leydig cells that produce testosterone are present in the interstitial tissues.

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

1°follicle → 2°follicle → 3°follicle → Graffian follicle → Corpus luteum

1.Cortex comprises corpus luteum along with mature follicles.
2.Inner medulla has blood vessels and degeneratin follicles
 EXPERMINT NO. :5

AIM: to study the T.S. of blastula through permanent slide.

Requirements: permanent slides of blastula, Microscope

T.S. of blastula through permanent slide (Mammalian).

COMMENTS:

1. 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.

2. Blastula appears as a sphere with a cavity known as blastocoel.

3. An outer layer of blastomeres known as trophoblasts is observed.

4. One end of the blastula shows a cellular mass adhered to the trophoblast.

EXPERIMENT NO.: 6

AIM: to study the meiosis in grasshopper testis through permanent slide.

Requirements: permanent slide of different stages of meiosis in grasshopper tesis , microscope.
Comments: It is the division of diploid germ cells that reduces the chromosome number by half resulting in the
production of haploid daughter cells (gametes).
- It occurs during gametogenesis.
- It ensures the production of haploid phase in the life cycle of sexually reproducing organisms whereas
fertilisation restores the diploid phase.

Key features of meiosis

 It involves two cycles called meiosis I & meiosis II but only a single cycle of DNA replication.
 It involves pairing of homologous chromosomes and recombination between them.
 Meiosis I begins after replication of parental chromosomes to form identical sister chromatids at the S phase.
 4 haploid cells are formed at the end of meiosis II.

Meiosis I Meiosis II
Prophase I Prophase II
Metaphase I Metaphase II
Anaphase I Anaphase II
Telophase I Telophase II

Meiosis I
1. Prophase I:
- It is the typically longer and more complex.
- It includes 5 phases based on chromosomal behaviour:
Leptotene, Zygotene, Pachytene, Diplotene & Diakinesis.

Leptotene:
 Chromatin fibres become long slender chromosomes. Nucleus enlarges.

Zygotene:
 Chromosomes become more condensed.
 Similar chromosomes start pairing together (synapsis) with the help of a complex structure called
synaptonemal complex.
 The paired chromosomes arecalled homologous chromosomes.
 Each pair of homologous chromosomes is called a bivalent
Pachytene:
 Comparatively longer phase.
 Bivalent chromosomes split into similar chromatids.Thisstage is called tetrads.
 This is characterised by the appearance of recombination nodules at which crossingover occurs.
Crossing over is the exchange of genetic material between non-sister chromatids of two homologous
chromosomes with the help of an enzyme, recombinase.
 It leads to recombination of genetic material on the homologous chromosomes. Recombination is
completed by the end of pachytene.

Diplotene:
 Dissolution of the synaptonemal complex occurs. The recombined homologous chromosomes of the
bivalents separate from each other except at the sites of crossovers.
 These X-shaped structures are called chiasmata.

Diakinesis:
 Terminalisation of chiasmata occurs.
 Chromosomes are fully condensed.
 The meiotic spindle fibres originate from the poles to prepare the homologous chromosomes for
separation. Nucleolus & nuclear envelope disappear.

2. Metaphase I:
 Spindle formation is completed.
 The chromosomes align on the equatorial plate. The microtubules from the spindle attach to the pair of
homologous chromosomes.

3. Anaphase I:
 The homologous chromosomes separate, while sister chromatids remain associated at their
centromeres.

4. Telophase I:
 The nuclear membrane and nucleolus reappear and 2 haploid daughter nuclei are formed. This is
called dyad.
 After this, cytokinesis may or may not occur. After a short interphase, it is followed by meiosis II.
 This short stage between the two meiotic divisions is called interkinesis. DNA replication does not occur
in this phase.

Meiosis II
It resembles the mitosis. It has the following phases:
 1. Prophase II:
 It is initiated immediately after cytokinesis.
 The chromosomes again become compact. Nucleolus and nuclear membrane disappear in both nuclei.

2. Metaphase II:
 The chromosomes align at the equator and the microtubules from opposite poles of the spindle get
attached to the kinetochores of sister chromatids.

3. Anaphase II:
 It begins with the simultaneous splitting of the centromere Meiosis II of each chromosome (which was
holding the sister chromatids together), allowing them to move toward opposite poles of the cell.

4. Telophase II:
 The two groups of chromosomes once again get enclosed by a nuclear envelope;
 cytokinesis follows resulting in the formation of tetrad of cells i.e., 4 haploid daughter cells.

EXPERIMENT NO.: 7

Aim: To perform emasculation, bagging and tagging for controlled pollination

Requirement: wild plants bearing large bisexual flower, magnifying lens, tweezers, small sharp scissors, brush, alcohol,
rubber bands, paper bags, paper clips and tags

Procedure :

(i) Emasculation: Select a flower in bud condition where antheses has not occurred. Open the bud carefully
and remove the stamens. Mark this as female parent plant.
 (ii) Bagging: Cover the emasculated flower with a plastic bag to protect it from undesired pollen (Bagging) .
The bag should be held securely in place with a paper clip/ string/thread. Select the size of bag in
accordance with the flower size. Bags must be transparent with minute pores.

(iii) Dusting : Bring into physical contact anthers of a desired male plant containing mature pollen grains with
the stigmatic surface of emasculated female flower . Use tweezers/brush if necessary to dust the stigmatic
surface with pollen.
(iv) Rebagging: Cover the pollinated flower again with the bag immediately. For identification, label the female
parent (Tagging). Each pollinated flower should bear a label containing the name of the seed parent, the
letter X (to signify a cross), the name of the pollen parent, and the date on which the cross was done.

EXPERIMENT NO.: 8
AIM: Prepare a temporary mount of onion root tip to study mitosis.
Principle: Onion root tip has meristemtic tissue . this tissue is responsible for growth of the onion root as the cells
of this region are highly dividing cells. They undergoes repeated mitosis division to increase the length of the
roots.

Mitosis division is also called the equational division as the chromosome numbers remain same in progeny cells
as present in parental cell. This division is required for growth, repairing of damaged tissue, replacement of dead
cells and for asexual reproduction.

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

Procedure:

# Growing of root tips:

i. Select medium-sized onion bulbs.

ii. Carefully remove the dry outer scaly leaves and roots present.
iii. Grow root tips by placing the bulbs on glass tubes (of about 3–4 cm. diameter) filled with water. Care
should be taken so that the stem portion of the bulb (basal part) just touches the water.
iv. Replace water in every 2-3 days.

# Fixation of root tips:

i. Cut 2–3 cm long freshly grown roots and transfer them to freshly prepared fixative, i.e., aceto-alcohol (1:3::
glacial acetic acid : ethanol).
ii. Onion root-tip cells have a cell cycle of approximately 24-hour duration, i.e., they divide once in 24 hours,
and this division usually takes place about two hours after sunrise.
iii. Therefore, roots grown on water should be cut only at that time to score maximum number of dividing
cells.

#Preparation of slide:

i. Take one preserved roots, wash them in water on a clean slide.

ii. Place one drop of N/10 HCl on the root tip followed by 2–3 drops of acetocarmine stain on it.
iii. Warm slide for 2-3 minutes slightly on spirit lamp).
iv. Care should be taken that the stain is not dried up. Carefully blot the excess stain using blotting paper.
v. 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.
vi. After (10–20 seconds) put one or two drops of water and blot them carefully using blotting paper.
vii. Again put a drop of water on the root tip and mount a cover slip on it avoiding air bubbles.
viii. 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.
ix. 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.
x. Carefully seal the margins of the cover slip using molten paraffin wax or nail polish. This preparation of
onion root tips cells is now ready for the study of mitosis.
xi. Study of slide 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:

1. Interphase:
i. The cells are mostly rectangular, oval or even circular in shape, with almost centrally situated densely
stained nucleus.
ii. The chromatic (coloured) material of the nucleus is homogeneous and looks granular. The boundary of
the nucleus is distinct.
iii. One or few nucleoli (sing: nucleolus) can also be observed inside the nucleus .
2. Prophase:
i. Intact nuclear outline is seen.
ii. The chromatin (seen as a homogeneous material in the nucleus at interphase) appears as a network of
fine threads (chromosomes).
iii. Nucleoli may or may not be visible.
3. Metaphase :
i. The nuclear membrane disappears.
ii. Chromosomes are thick and are seen arranged at the equatorial plane of the cell.
iii. Each chromosome at this stage has two chromatids joined together at the centromere.
4. Anaphase:
i. This stage shows the separation of the chromatids of each chromosome.
ii. The chromatids separate due to the splitting of the centromere.
iii. Each chromatid now represents a separate chromosome as it has its own centromere.
iv. The chromosomes are found as if they have moved towards the two poles of the cell.
5. Telophase :
i. Chromosomes reach the opposite poles, lose their individuality, and look like a mass of chromatin.
ii. Nuclear membrane appears to form the nuclei of the two future daughter cells. (one cell two nuclei)

Precautions:

1. The base of the onion bulb should be n contact with water while growing the roots.
2. Clean the slide and coverslip thoroughly before use.
3. Avoid air bubbles while putting coverslip on the slide.
4. Root tips should be fixed in the morning between 8 to 10 am.
5. The slide should be warmed gently much above the flame of the spirit lamp
 EXPERIMENT NO.: 9
Aim: Isolate DNA from available plant material such as spinach, green pea seeds, papaya, banana, Cauliflower etc.

Principle: DNA is one of the nucleic acids found in living organisms. DNA acts as the genetic material in organisms
except some virus. rDNA technology involves isolation of DNA from a variety of sources and formation of new
combination of DNA. DNA can be extracted from the cell by degrading cell wall, plasma membrane and nuclear
membrane ( using liquid soap and NaCl) followed by precipitation of (DNA using ethanol)

Requirements: Plant material (banana), Water, Pastel and mortal or grater, Chilled Ethanol (Refrigerate it overnight),
NaCl, Liquid detergent, Muslin cloth for filtration, tooth pick, Large paper clips/ Wire loop, Beaker, Petri dish, Boiling
tube

Procedure:

1. Take the available plant material and grind it in the mortar or grate/mesh it to make paste in a petri
dish/beaker.
2. Fill a clean beaker with 25 ml of water, slowly add one teaspoons of liquid detergent and half teaspoon of
NaCl.
3. Gently mix tem without making bubbles till the salt dissolves.
4. Add this mixture to meshed plant material and let it undisturbed for 20 minutes to give detergent enough
time to react.
5. Place a fine/muslin cloth on a small beaker/boiling tube and carefully pour the mixture here and filter it.
6. Gently squeeze the mixture to get more liquid out. This liquid filtrate contains DNA.
7. Since the DNA is soluble in water so to isolate DNA from this filtrate pour chilled ethanol by side of slightly
(450) tilted boiling tube.
8. After few minutes DNA will isolate as white precipitates/ fine threads from the watery filtrate at the boundary
layer between water and ethanol.
9. Separate DNA by spooling i.e. the winding of the fine threads of DNA on clip or wire loop.

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

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

Precautions: 1. All the glass wares must be thoroughly cleaned and dried.

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

3. NaCl and Liquid detergent should be to dissolve slowly by stirring without formation of foam or bubbles.

4. Add chilled ethanol to enable the precipitation of the DNA

5. Use wire or blunt forceps for spooling of precipitated DNA.

 Experiment no.: 10
AIM: Common disease causing organisms like Ascaris, Entamoeba, Plasmodium,
any fungus causing ringworm through permanent slides or specimens. Comment
on symptoms of diseases that they cause.

Requirements: Permanent microscopic slides of the Entamoeba hystolytica, Plasmodium

vivax and specimen of Ascaris lumbricoides

Ascaris lumbricoides
COMMENTS:
Phylum – Aschelminthes (Round worm)
Class – Nematoda
Type – Ascaris lumbricoides
External features:
1. It has a long, cylindrical and un segmented body.
2. The male and female organisms are separate.
3. It bears a mouth at the anterior end surrounded by three lips.
4. There is an excretory pore on the ventral surface slightly behind the anterior end.
5. A pair of penial spicules are present in the male worms close to the cloacal opening.
6. The female genitals are present at about one-third distance from the anterior end.
7. Disease Caused In Humans: It is one of the common parasite found in the intestine of human
beings that causes Ascariasis.
8. Comon symptoms of disease: Symptoms of these disease include
a. internal bleeding,
b. muscular pain,
c. anemia and
d. blockage of the intestinal passage.
e. fever,
9. Spreading of disease: The eggs of the parasite are excreted along with the faeces of infected
persons which contaminate soil, water, plants, etc. A healthy person acquires this infection
through contaminated (eggs present) water, vegetables, fruits, etc

Plasmodium vivax
External Features:
1. It is a unicellular endoparasite found within the red blood cells of the diseased person.
2. The parasite is mostly diagnosed at the “signet ring” stage where the parasite appears as a
round body.
2.
 3. 3. There is a big vacuole present inside the cell. The cytoplasm is accumulated at one place and
contains the nucleus.
4. 4. Plasmodium vivax is a protozoan parasite that causes malaria in humans. The infected female
anopheles bites a healthy person and transmits the sporozoite into the peripheral blood vessels of
humans.

Disease: The infective stage sporozoites causes the disease Malaria. This stage undergoes several rounds of
multiplication in liver and erythrocytes of Human.

Symptoms:
1. chill and high fever recurring every three to four days
2. Shaking chills,
3. Headache,
4. Vomiting,
5. Nausea

SPREADING OF DISEASE:
Get spread due to bite of female Anopheles mosquitoes.

Entamoeba histolytica
External features:
1. It is a unicellular organism with an irregular shape.
2. It consists of a few food vacuoles.
3. The contractile vacuole is absent.
4. Cysts with four nuclei are present.
5. It consists of a nucleus located eccentrically in the cell.
Disease:
1. Entamoeba histolytica is an organism found in the intestines of humans that is responsible for
causing amoebic dysentery.
Symptoms:
1. constipation,
2. abdominal pain and cramps,
3. stools with excess mucous and
4. blood clots.

SPREADING OF DISEASE:
1. Houseflies act as mechanical carriers and serve to transmit the parasite from faeces of infected
person to food and food products, thereby contaminating them.
2. Drinking water and food contaminated by the faecal matter are the main source of infection
 EXPERIMENT NO. 11

AIM: Study the plant population density by quadrat method.

Principle:

1. The number of individuals of the species in any unit area is its density. The unit area may be
as small as 5 square cm to as large as 10 square metre.
2. For herbaceous vegetation a metre square quadrat is normally used. Density which gives an
idea of degree of competition is calculated as follows.
Density= Total number of individual(s) of the species in all the sampling unit
Total number of sampling units studied (Q)

Requirement:

Meter scale, Cotton/nylon thread (five meters), 4 nails and a hammer

Procedure:
1. In the selected site of study, make a 1 m X 1 m quadrat with the help of nails and thread.
Hammer the nails firmly and make sure that the vegetation is not damaged while laying the
quadrat.
2. List the names of the plant species seen in the quadrat (if the name is not known mark these
as species A or B etc., and the same species if seen in other quadrats assign the same
alphabet).
3. Count the number of individuals of each species present in the quadrat and record the data as
shown in the table.
4. Similarly make nine more quadrats randomly in the site of study and record the names and
number of individuals of each species.
I WILL GIVE A PAPER TO BE PASED ON LEFT SIDE OF PROCEDURE SO PLEASE LEAVE
THE COMPLETE PLAIN PAGE AT LEFT

Observations:

Record the total number of species seen in the ten quadrats.

(DRAW THE TABLE AT LEFT page with pencile . Same table drawn by students in the files

Conclusion

The population density is the highest for species …......... and the lowest for species …........... The
density value is expressed as the number of individuals per unit area.
Precautions:

1. Measure the quadrate accurately.

2. Mark all the quadrates close to each other within one field only.
3. The string/ thread should not be very tick.
4. Every individual of all species should be counted precisely without repetition. 5. The vegetation
should not be damaged while laying the quadrates.

EXPERIMENT NO.:12

Aim:Study the plant population frequency by quadrat method.

Principle:

1. Plant population frequency is” the number of times a species of plant is repeated in a
specified quadrat”.
2. The frequency of each species (sps. A or sps. B or sps. X etc) is expressed in percentage
and is calculated as follows.
3. percentage frequency= no. of sampling units (quadrats)in which the species occurs
total no. of sampling units (quadrates)

Requirements:
Meter scale, Cotton/nylon thread of 5 metres, 4 nails and a hammer

Procedure:

10. In the selected site of study, make a 1 m X 1 m quadrat with the help of nails and thread.
Hammer the nails firmly and make sure that the vegetation is not damaged while laying the
quadrat.
11. List the names of the plant species seen in the quadrat (if the name is not known mark these
as species A or B etc. and if the same species is seen in other quadrats assign the same
alphabet)
12. Similarly lay nine more quadrats randomly site of study and record the names of individuals of
each species.
13. Calculate the percentage frequency of occurrence using the formula given.
LEAVE COMPLETE PAGE LEFT SIDE OF PROCEDURE. I WILL GIVE A PAGE NEED TO
BE PASTED AT LEFT

Observations:

1. Record the total number of species seen in the ten quadrats. This will give an idea about
the composition of the vegetation.
Observe that the frequency of occurrence is not the same for all species. (There will be difference in
the species composition (DRAW THE TABLE AT LEFT page with pencile . Same table drawn by
students in the files

Conclusion

The plant population frequency is the highest in species species ...........

Precautions:

1. Measure the quadrate accurately.

2. Mark all the quadrates close to each other within one field only.
3. The string/ thread should not be very tick.
4. Every individual of all species should be counted precisely without repetition.
5. The vegetation should not be damaged while laying the quadrates.