BATCH I

1. Isolate DNA from Available Plant Material Such as Spinach, Green Pea Seeds, Papaya

Aim
To isolate DNA from available plant material like pea seed, spinach, onion, etc.
REQUIREMENTS
Plant material (like pea-seed, spinach leaves, onion or cauliflower shavings), mortar and pestle, beakers, test
tubes, enzymes (like cellulase, protease, lipase and ribonuclease), ethanol, spool, glass rod, etc.
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 hydrolyse 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
Shiny white DNA can be seen at the junction of solution and ethanol. This DNA represents all the DNA found
in plant cells. The chromosomes were broken in the process and the DNA gets precipitated due to the chemical
treatment

RESULT
DNA appears as white precipitate of very fine threads on the spool
PRECAUTIONS
1.Fresh plant materials should be used for extraction of DNA. 2.The glassware must be thoroughly cleaned and
dried before the experiment starts and always use distilled water to make the solutions. 3. Chemicals and
solutions should be prepared carefully in order to avoid any wastage. 4. The enzymes and chemicals used for the
experiment should be of standard quality.
2. Study the Plant Population Frequency by Quadrat 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 x 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, C, D, and E in all the quadrants and mention them in the
table.
8. Determine the frequency of plant population by the formula:

Plant Number of Quadrats Employed in the Total Number of Percentage of

quadrats in which frequency
Species Study (Q) Number
quadrats The species is F=N/Q x 100
(Q) present (N)

I II III IV V VI VII VIII IX

A 2 1 3 1 1 2 2 1 1 9 9 9/9x100 =100%

B 1 1 1 1 1 0 0 0 0 9 5 5/9x100 =55.5 %

C 0 0 0 1 1 1 1 2 1 9 6 6/9x100 = 66.6 %

D 0 1 2 0 1 0 0 0 0 9 3 3/9x100 = 33.3 %

E 1 2 1 1 1 2 1 1 2 9 9 9/9x100 = 100 %

Result:
The plant population frequency is the highest in species A and E and the least in species D. It shows how many
times a plant species is present in the provided number of sample quadrats
PRECAUTIONS
The measurement of quadrats should be accurate.
The string or cord used should not be very thick.
One individual of a species should be counted only once in the quadrat.
Make sure that the vegetation is not damaged, while laying the quadrat.
3. Prepare A Temporary Mount of the 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, WaterN/10 Hydrochloric acid,
Acetocarmine Stain, Burner, Forceps, Dropper, Blade, Needle, Compound microscope

Procedure

 Grow the onion 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 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
Different stages of mitosis, prophase, metaphase, anaphase, telophase and cytokinesis were observed in onion
root tip cells.
Result.
Metaphase is the second step of mitotic cell division after prophase. Metaphase is characterised by the
alignment of the duplicated chromosomes at the equatorial plate known as the metaphase plate. One of the sister
chromatids is attached to one pole of the cell by microtubules attached to its kinetochore and the other sister
chromatid gets attached to the opposite pole.

PRECAUTIONS

Always clean the slide and coverslip thoroughly before and after use.

Care should be taken that there should be no air bubbles under the coverslip.

Always filter the acetocarmine stain before use.

Material should be mounted on the centre of slide.

SPOTTERS

4. The given spotter is identified as T.S (transverse section) of Mammalian Testis.

Identification Characteristics* The seminiferous tubules are easily seen in cross-section of testis.

* The presence of larger cells called Sertoli cell is the characteristic feature of this slide.

* The presence of interstitial cells or Leydig's cell (secrete testosterone) is evident.

General Characteristics

* The transverse section of testis shows many seminiferous tubules embedded in interstitial tissues. These
appear as coiled, circular structures.

* Each seminiferous tubule is lined on its inside by germinal epithelium which divides mitotically to produce
male germ cells called spermatogonia.

* Various stages of development of sperm from spermatogonia are seen from periphery to the lumen of
seminiferous tubules. The sequence of stages of its development is as follows:

5. The given spotter is identified as Ornithophilous flower – Bird pollinated flower – Bignonia capreolata.

 The flowers are big and brightly coloured red, orange, yellow or blue.
 The flower parts are thick and leathery especially corolla.
 The flowers produce abundant nectar and may also have certain edible parts. Birds visit flowers for
feeding on this nectar.
 The flowers are usually odourless (without any fragrance or scent).
6. The given spotter is identified as Anaphase 1 of meiosis 1
*In anaphase I, the homologues chromosomesare pulled apart and move apart to opposite ends of the cell.

* The centromere does not divide.

*Each homologous pair of chromosomes moves to the pole with both the chromatids.

*The sister chromatids of each chromosome, however, remain attached to one another and don't come apart

7.The given spotter is identified pedigree chart forRolling of Tongue:

* It is an autosome linked dominant trait.* The tongue rolling ability is present in some (dominants) while in
others it is absent others (recessive). Such individuals are called tongue – rollers and tongue non rollers
respectively

* The trait is represented by dominant gene (RR homozygous dominant /Rr heterozygous dominant) and
recessive gene (rr)

It is not a sex-linked trait.In this pedigree chart both the parents are heterozygous dominant for rollers (Rr and
Rr).Out of their three children one boy (Rr)and one girl (Rr)roller and one girl (rr) is non-roller of tongue

8. The given spotter is identified as disease-causing organismAscaris lumbricoides

Ascaris is an endoparasite of small intestine of human beings.It causes the disease Ascariasis

Symptoms.: - Ascaris worm infests a single host and obstructs the intestinal passage thereby causing
abdominal discomforts.

 The sufferer (infected person) complaints of colic pain.

 There is impaired digestion, vomiting and diarrhoea in severe infestation.
  As an endoparasite it uses all nutrients and the infected person shows symptoms of malnourishment
like anaemia, retarded body growth and mental growth specially in children.
 Person loses weight and feels tired and lethargic.

9. The given spotter is identified as Homologous organ

 Homologous organs are referred to as the organs/traits that are inherited by two different organisms but
from the same ancestry.
 They are similar in morphology but their functions are different.
 They have the same embryonic origin.
 Homology shows the divergent evolution that is the one ancestry that leads to further development in
the organisms.
 Examples of homologous organs are the forelimbs of human, cat, bat and the flippers of whales.

10.. The given spotter is identified as Scissors/Forceps Method of Emasculation

This is one of the methods for controlled pollination.

(i) This method is adopted for the plants having large sized flowers, e.g., cotton, pea, etc.
(ii) (ii) Emasculation is done with the help of pocket lens, forceps, needle, scissors, scalpel camel hair
brush as shown in
(iii) The anthers are cut in the flower before their maturation with the help of sterilised forceps or
scissors. This is called female flower now.