DNA ISOLATION
Aim
To isolate DNA from plant materials such as spinach, green peas, papaya and any
other available plant material.
Necessary Materials & Apparatus
Any available plant materials
Mortar and pestle
Test tubes
Beakers
Ethanol
Spool
Enzymes (Cellulase, ribonuclease, lipases, protease)
Procedure
Take the available plant material and grind it in the mortar.
Treat the material with cellulase to break down the cell wall of the plant cells.
Next, treat it with protease to hydrolyze the peptide bonds of proteins in the plant
material. In other words, the enzyme removes the histone proteins which are intertwined
with the DNA.
Dissolve RNA with ribonuclease
Use lipase to dissolve lipids.
Add chilled ethanol to enable the precipitation of the DNA. It essentially increases DNA
concentration.
Use spooling to extract the precipitated DNA. Spooling involves winding the fine threads
of DNA on to a reel.
Observation
The DNA appears as white precipitates of fine thread on the spool.
Pollen Tube Germination
Aim
To study pollen germination on a slide.
Necessary Materials & Apparatus
Freshly plucked seasonal flowers, beaker, boric acid, sucrose, microscope and
cavity slide.
Procedure
The first step involves the preparation of a nutrient solution. This is done by
dissolving 10g of sucrose as well as 10mg of boric acid in 100ml of water.
Pour a few drops of this solution onto the cavity slide. Then, use a brush or fingers to
gently dust a few pollen grains from the stamen of mature flowers.
Let the slide set for 5 mins. Then, use the microscope to view the slides in 30-minute
intervals.
Observation
The pollen grains will germinate when submerged in the nutrient-rich medium. This is
characterized by the enlargement of the vegetative/tube cell. It emerges through one
of the germ pores, eventually forming a pollen tube. The generative cell nucleus
grows into the pollen tube and makes two male gametes (sperm nuclei). The male
gamete is either spherical or lenticular in outline.
Precautions
Ensure that the flowers are freshly picked
The observation slide should be a cavity slide, meaning that it has a depression in the
centre.
TS OF OVARY AND TESTIS
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.
TS OF BLASTULA
Aim
To observe the TS of mammalian blastula through permanent slides.
Materials Required
Permanent slide
Compound microscope
Procedure
Observe the permanent slide under a microscope and note down the features of TS
of the mammalian blastula.
Observations
Blastula appears as a sphere with a cavity known as blastocoel. An outer layer of
blastomeres known as trophoblasts is observed. One end of the blastula shows a
cellular mass adhered to the trophoblast. This is known as the inner cell mass
Emasculation and Bagging
Aim
To control pollination through emasculation, tagging and bagging.
Materials Required
Plants with large bisexual flowers
Tweezers
Scissors
Brush
Alcohol
Rubber bands
Paper bags
Paper clips
Tags
Magnifying Glass
Theory
Emasculation is the process of artificial hybridization in which the stamens from the
female flowers are removed from bisexual flowers in order to prevent self-
fertilization. This process is carried out long before the anthers mature. Removal of
anther from the bisexual flowers before the anthers mature is known as
emasculation. The emasculated flower is then bagged to prevent any unwanted
pollination.
This process helps in the production of flowers with desired characteristics. For this,
it is essential to have knowledge of the flower structure, fertilization, physiology of
flowers and fertilization.
Procedure
1. Select a bud of the flower and open it to remove the stamens. This is known as
emasculation. Mark it as the female parent plant.
2. The plant is then covered with a plastic bag to prevent it from getting pollinated by any
undesired pollen.
3. Bring it in contact with the anther of the male plant with desired characteristics. The
pollen should be dusted on the surface of the stigma.
4. Cover the pollinated flower immediately with a polythene bag and label it with the name
of the seed parent.
Population Density
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:
𝐷𝑒𝑛𝑠𝑖𝑡𝑦=𝑇𝑜𝑡𝑎𝑙 𝑛𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑖𝑛𝑑𝑖𝑣𝑖𝑑𝑢𝑎𝑙𝑠 𝑜𝑓 𝑡ℎ𝑒 𝑠𝑝𝑒𝑐𝑖𝑒𝑠 𝑖𝑛 𝑎𝑙𝑙 𝑠𝑎𝑚𝑝𝑙𝑖𝑛𝑔 𝑢𝑛𝑖𝑡𝑠(𝑆)
𝑇𝑜𝑡𝑎𝑙 𝑛𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑠𝑎𝑚𝑝𝑙𝑖𝑛𝑔 𝑢𝑛𝑖𝑡𝑠 𝑠𝑡𝑢𝑑𝑖𝑒𝑑(𝑄)
D = S/Q
Observations
Plant Number of individuals in each Total number of Total number of Density
Species quadrant individuals (S) quadrants (Q) (D)=S/Q
I II III IV V VI VII VIII IX X
A 2 0 5 7 10 0 0 0 0 3 27 10 2.7
B 1 0 4 0 8 0 3 0 0 2 20 10 2
C 4 0 0 3 0 6 0 0 1 2 19 10 1.9
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.
Population Frequency
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:
Percentage frequency= (No. of sampling units in which species occur)/(Total
number of sampling units used in the study)*100
Observations
Plant Quadrants employed in the No. of quadrants in which Percentage
Species study species are present (N) Frequency
(F)=N/Q*100
I II III IV V VI VII VIII IX X
A P P P 3 30%
B PP P P4 40%
C PP P PP 5 50%
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.
