EXPERIMENT 8.

AIM: To isolate DNA from available plant material such as spinach leaves, green pea seeds,
papaya etc.

REQUIREMENTS
Plant material (such as
green pea seeds or green papaya), mortar and pe
spinach leaves, or
beakers, test tubes, liquid detergent, non-iodised sodium chloride, distilled water, meat tendere
papain solution/juice of papaya/pine apple juice, 95% ethanol, spool etc.

PREPARATION OF SOLUTIONS
dised
e e r g e n t salt solution is prepared by adding 10 mL liquid detergent and 10 g or "no
sodium chloride to 90 mL of distilled water. istilled
Meat
Water
tenderizer solution is prepared bv adding 5 g of tenderizer (enzyme) to 95 n .
(Juice of
papaya/pine apple, filtered through muslin cloth can
meat tenderizer).
be used ds

S% NaCl solution is prepared by dissolving chloride in 100

mL
distilled water. 5 g of non-iodised sodium cno
over
c l a n o l must be done
night. by keeping 95% ethanol in plastic bottle in the rree
 55
Core Experiments

PROCEDURE

it in the
. Take 5 g of the plant tissue (spinach leaf/greenpea seed/green papaya) and grind
salt solution and filter it through muslin cloth.
mortar by adding 10 mL detergent,
tube by holding
Take 10 mLof the filtrate, add 3-4 mL tenderizer/papaya juice and swirl
the test

to mix the contents.

the tube between the two hands
down the side of test tube to form layer on the top of
Pour 10 mL chilled ethanol carefully
a

the content; let it stand undisturbed for about 3 minutes.

interface of the two layers to collect the precipitate of
.Using the glass rod stir gently through
DNA and place it in a test tube with 5% NaCl or distilled water.

of DNA present in the given plant material can be estimated through

The quantity
spectrophotometer.

Fig. 8.1. DNA that separates out can be removed by spooling (spool =reel for winding yarn).

OBSERVATION
The addition of ethanol to the solution causes DNA to precipitation. The DNA fibres appears as

white precipitate of very fine threads on the glass spool.

PRECAUTIONS
distilled water to remove any dust and
1h e plant material should be washed throughly with
dried by blotting before weighing&
cleaned and dried.
used be thoroughly
Al
the glasswares must
be of standard quality which should
must
3. ne chemicals and enzymes used for the experiment
be manufactured by standard pharmaceuticals.
 EXPERIMENT 9.1

AIM: To study the flowers adapted to pollination by different agencies (wind, insect and birds).

REQUIREMENTS
Fresh flowers of maize or any other cereal/grass, Salvia/Ocimum and Brassica (mustard) forceps,
hand lens, slide, needle etc.

PROCEDURE
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 agencies.
Maize Flowers (Anemophilous or Wind Pollinated Flowers)
The flowers of maize show
following adaptations for pollination by wind.
1. The maize plant is monoecious
and bears unisexual flowers. The male flowers are bornin te
inflorescence while the female flowers born in
2.
are
axillary inflorescence.
Flowers are small and
3 The flowers
inconscipicous.
are
colourless, odourless and nectarless.
4. Flowers are
produced above the foliage or placed in hanging position
5. Both the
stigmas and anthers are exerted (i.e., hang outside the
6. Anthers are versatile, and perianth).
pollen grains are light, small and
7. The
pollen grains are dusEy
produced in very large numbers.
Core Experiments
59
8. Stigma is hairy, feathery or branched to catch wind born
pollen grains.
Male flowers

Tassel

Pollen grains

Cob
Versatile
Elongated anther
styles Feathery
Pollen grains stigma
of another plant
Stigmas Ovary

Fig. 9.1. Anemophily in maize. Fig. 9.2. Feathery stigmas and versatile
anthers in a flower of grass.

Salvia Flowers
(Entomophilous or Insect Pollinated Flowers)
The flowers of Saliva show
following adaptations for pollination by insects.
1. The flowers are
showy
or brightly coloured for attracting pollinating insects.
2. The flowers are born in verticellaster inflorescence to become
3 conspicuous.
Flowers secrete nectar to feed
an insect must
visiting insects. Nectar glands are placed in such a position that
touch both the anthers and stigmas.
4 The flowers have
landing platform for the insects.
5. The flowers are protandrons with
bilipped corolla and have turn pipe or lever mechanism.
6. Each stamen has
long connective which bears a fertile anther lobe at the
plate like anther lobe at the lower end. The two sterile upper end and sterile
anther plates block the
7. As the insect moves path of insect.
inward a young flower in search of nectar, its head
and forces the fertile pushes, the anther plates
anther lobes to strike its back.
against
8. In older flowers the
of insect and
style brings the stigma in such a position that it brushes against the back
collect pollen grains brought by the insect from a young flower.
 amprehenstue Laboratory Manual in Biolog
60
Closed stigma Fertile
anther lobe

Shedding of
pollen grains
on the back
of insect

3
Sterile
A anther lobe Stigma
Nectariferous
Mature receiving
disc /stigma pollen grains
from the back
of insect

D
C
Withering
anther
and short style.
Fig. 9.3.ofPollination in Salvia. A. Flower with mature anthers, enclosed stigma
B. Shedding pollen grains on the back of entering insect. C. Flower with mature stigma and withering
anthers. D. Stigma receiving pollen grains from the back of entering insect.

or Bird Pollinated
Bignonia/Callistemon (Bottle brush) Flowers (Ornithophilous
Flowers)
The flowers of Bignonia show following adaptations for pollination by birds.
The flowers are usually brightly coloured-red, orange, yellow or blue.

2. The floral parts are commonly leathery.

w.

Humming bird

Fig. 9.4. Pollination in Bignonia. Humming bird collecting nectar from

Bignonia flower and thus pollinating it.
Core E x p e r i m e n t s 61

the corolla are leathery.

3 In some cases,

abundant watery nectar or have edible parts.

The flowers secrete
4.
The nectar is secreted in such abundance that drops of it can be brought down by shaking
5
branches.
without fragrance.
The flowers are generally odourless or
6.
 EXPERIMENT 11.1
AIM: To study and identify the stages of gamete development in mouse (mammal) i.e., T.S. of
testis and L.S. of ovary through permanent slide.

REQUIREMENTS
Permanent slide of T.S. of testis and L.S. of ovary, microscope.

PROCEDURE
FIx the permanent slide under the microscope. First observe it under the low power and then
under high power.

OBSERVATION
T.S. of Testis
n etestis

2
of a mouse (mammal) is covered by a thick fibrous tissue called tunica albuginea.

3 SEIS consists of numerous seminiferous tubules embedded in the interstitial tissue.

Various typ
ypes of germinal cells are present from outside towards lumen in the following sequence

4 natogonia > Spermatocytes Spermatids-> Spermatoz0a >Sperms

Between the
en the germinal
ger cells, pyramid shaped cells called sertoli cells are present.
66 Comprehensive Laboratory Manualin
with their heads embedded in
Biolo
dogy-
sertoli
5. A large number of spermatozoa cells are
rell.

lumen of seminiferous tubule. presenttintin

6. The interstitial tissue
also contain leydigs cells, which produce male sex male sex hor
hormone
testostero
Visceral
peritoneum
Tunica
albuginea

Blood vessel Spermatozoa

Seminiferous Spermatid
tubule
Secondary
spermatocyte
Sertoli cell- Primary
Connective
spematocye
tissue Ooo
Sertoli cel
Germinal epithelium
Spermatogonia Spermatogonin
Spermatocytes
Spermatids
Spermatozoa
Interstitial
cells B
A

Fig. 11.1.A. A Part of transverse section testis of mouse (mammal)

of
seminiferous tubule (enlarged)
B. Sectional view of a part of

V.S. of Ovary
followed by a thick layer dt
1. A mouse ovary is a solid structure bounded by germinal epithelium
fibrous tissue, the tunica albuginia.
Egg nest Primary follicle
Cortex Secondary
follicle
Blood
vessel Te tiary
follicle

Visceral

Mesovarium peritoneum

Graafian follicle
Corpus
albicans

Corpus luteum

Ruptured follicle
Medulla

Fig.11.2. A section of ovary of mouse (mammal).

Core Experiments 67

2. The ovary consists of outer cortex and inner medulla.

3. The medulla contains many rounded or oval bodies called ovarian or Graafian follicles at various
stages of development.
fibres and smooth muscles.
4 The medulla also contains blood vessels, nerves some

5. Each follicle contains a large ovum surrounded by many layers of follicle cells.
6. The cortex contains young and mature follicles.

7. The cortex also contain a large m a s s of yellow

cells termed corpus luteum, formed in an
may
empty Graafian follicle after the release of
its ovum.

PRECAUTIONS
under low and then under the high power of the microscope.
irst observe the slide power

for focussing the slide under high power.

se n e adjustment of the microscope
 EXPERIMENT 13.1

AIM: To study T.S. ofblastula through permanent slide.

REQUIREMENTS
Permanent slide of blastula, microscope.

PROCEDURE
Fix the slide of T.S. of blastula under microscope. First observe the slide under low power ane

then under high power of the microscope.

Inner cell mass
(Formative cells)

Trophoectoderm

Trophoblast),

Blastocoel
Zona
pellucida
Albuminous layer
around the zona

Fig.13.1. T.S. of blastula.

Core CAH

O B S E R V A T I O N S

spherical mass
of about sixty four cells,
1. It is a

Te comDosed of an outer envelope of cells, the trophoblast or trophoectoderm and inner cell
2. mass(= embryoblast).

there is fuid filled

cavity called
blastocoel.
Within the envelope
a
3.
to which the inner cell mass is attached is called the embryonic
The side of the blastocyst
or

animal pole, while the opposite side is the abembryonic pole.

The inner cell mass is the precursor
of the embryo.
5.

PRECAUTIONS

1 First focus the slide under low power and then under the high power of the microscope.

2. Use fine adjustment while focussing the slide under high power of the microscope.