BOTANY PRACTICAL

LAB MANUAL

Prepared By
SUBRAHMANIAN. P.M
HSST BOTANY
GOVT. HSS NARIKKUNI
Sl Page
No. Date
No.
FIRST YEAR PRACTICAL
CYTOLOGY- CELL DIVISION
1
PROPHASE
2 METAPHASE

3 ANAPHASE

4 TELOPHASE

MONERA
5 OSCILLATORIA
FUNGI
6
RHIZOPUS
7 AGARICUS - BASIDIOCARP
ALGAE
8 SPIROGYRA- VEGETATIVE FILAMENT

9 SARGASSUM- THALLUS
BRYOPHYTE
10 FUNARIA- GAMETOPHYTE WITH SPOROPHYTE

11 FUNARIA - PROTONEMA
PTERIDOPHYTE
12 NEPHROLEPIS/FERN - SPOROPHYTE

13 NEPHROLEPIS/FERN - PROTHALLUS
GYMNOSPERM
14 PINUS- MALE CONE

15 PINUS – FEMALE CONE

LICHEN
16
USNEA
Sl
Title Date Page No.
No.
SECOND YEAR PRACTICAL
ANATOMY OF ANGIOSPERMS
17
DICOT STEM.T. S
18 MONOCOT STEM. T. S

19 DICOT ROOT. T. S

20 MONOCOT ROOT. T. S
TAXONOMY
21 OBSERVATION OF FLOWER TO DESCRIBE FLORAL WHORLS
- SOLANACEAE – e.g; CAPSICUM
22 PREPARATION OF ANTHER.C. S
ECOLOGY- ECOLOGICAL INTERACTIONS
23
MUTUALISM - LICHEN
24 COMMENSALISM - VANDA

25 PARASITISM- CUSCUTA
BIOTECHNOLOGY
26
CLONING VECTOR pBR322
27 BIOREACTOR

28 BT. COTON

FIRST YEAR PRACTICAL

PHYSIOLOGY EXPERIMENTS
29
STUDY OF DISTRIBUTION OF STOMATA ON UPPER AND LOWER SIDES OF LEAVES
30 STUDY OF PLANT PIGMENT BY PAPER CHROMATOGRAPHY

31 HYDRILLA EXPERIMENT

32 YEAST FERMENTATION AND PRODUCTION OF ALCOHOL

 CELL DIVISION

STAGES OF MITOSIS

1. Prophase

Identification
The given stage is prophase of mitosis
Reasons
Chromosomes are thickened and distinct.
Spindle fibres start to originate.
Nuclear membrane and nucleolus start disappear

2. Metaphase

Identification
The given stage is metaphase of mitosis.
Reasons
Chromosomes are arranged at the equator of the cell.
The spindle fibres become attached at the centromere of chromosome
The arms of the chromosomes are directed towards the opposite poles.
3. Anaphase

Identification
The given stage is anaphase of mitosis

Reasons
The chromatids separate.
Chromosomes move to opposite poles of the cell.
The arms of the chromosomes are directed towards the equator of
poles.

4.Telophase

Identification
The given stage is telophase of mitosis.

Reasons
The two sets of chromosomes reach at opposite poles of the cell.
The chromosomes are arranged and form two daughter nuclei.
Nucleus and nuclear membrane reappear.
 3. OSCILLATORIA

Identification

The given slide shows filaments of Oscillatoria

Identification Features

It is freshwater, free living, nitrogen fixing cyanobacterium.

The filament is long and unbranched.
Filaments show oscillating movement.
 FUNGI

1. RHIZOPUS

Identification
The given slide shows mycelia with sporangia of Rhizopus

Identification Features
The fungal mycelium is coenocytic.
The mycelium is made up of hyphae; it consists of rhizoids, stolon and
sporangiophores.
Spores are produced inside the sporangium.
 2. AGARICUS

Identification

The given material is Agaricus

Identification Features

It is the fruiting body of Agaricus and is known as basidiocarp.

It consists of a stipe, and a cap.
Several gills are present at the lower side of cap and which produce
basidiospores.
 ALGAE
1. SPIROGYRA

Identification

The given slide is the Spirogyra filament

Identification Features

Spirogyra is a filamentous fresh water green alga.

The filament is cellular and unbranched.
Each cell consists of a ribbon shaped chloroplast.
 2. SARGASSUM

Identification

The given material is Sargassum alga.

Identification Features

It is a brown alga.
The thallus consists of hold fast, stalk and front.
The front consists of primary laterals secondary laterals and receptacle.
 BRYOPHYTES

FUNARIA

Identification

The given material is Funaria

Identification Features
The plant body consists of gametophyte and sporophyte.
The gametophyte of Funaria consists of root like, stem like and leaf like
parts.
The sporophyte is differentiated in to foot seta and capsule.
 FUNARIA – PROTONEMA

Identification

The given material is the protonema of Funaria

Identification Features

The spore of Funaria germinates and produces protonema.

The protonema is a green photosynthetic multicellular branched
structure with rhizoids.
Certain regions of the protonema produce buds and which develop into
erect gametophyte.
 PTERIDOPHYTES

NEPHROLEPIS

Identification

The given material is Nephrolepis sporophyte.

Identification Features

The sporophyte of Nephrolepis is differentiated into roots, rhizome, and

leaves.
The leaves are pinnately compound.
The leaves bear sporangia and are called sporophylls.
 FERN – PROTHALLUS

Identification

The given slide shows the prothallus of Nephrolepis.

Identification features

Prothallus of Nephrolepis is green, flat, heart shaped structure

germinated from the spore.
It is the haploid photosynthetic gametophyte.
Its lower side bears rhizoids, antheridia and archegonia.
 GYMNOSPERMS

PINUS - MALE CONE

Identification

The given material is the male cone of Pinus

Identification Features

The male cone consists of an axis on which the microsporophylls are

arranged spirally.
Each microsporophyll bears two microsporangia on the lower side.
Pollen grains are produced in microsporangia.
 PINUS - FEMALE CONE

Identification

The given material is the female cone of Pinus

Identification Features

The female cone has an axis on which the megasporophylls are spirally
arranged.
Each megasporophyll has two ovules at the base.
 LICHEN – USNEA

Identification
The given material is a lichen.

Identification Features
Lichen is the association of fungi and algae.
Fungal partner is called mycobiont and the algal partner is called
phycobiont.
 ANATOMY OF ANGIOSPERMS

DICOT STEM.T. S
Ground plan

Identification features of Stem

1.Presence of conjoint collateral vascular bundles.
2.Xylem is endarch.

Identification features of Dicot

1.Vascular bundles arranged in a ring.
A portion enlarged
2.Vascular bundles are open.
3. Ground tissue is differentiated into cortex and pith.
4. Limited number of vascular bundles.

Identification
So, the material is dicot stem.
 MONOCOT STEM.T. S

Ground plan
Identification features of stem
1.Vascular bundles are collateral and conjoint.
2.Xylem is endarch.
3.Sclerenchymatous hypodermis present.

Identification features of monocot

1.A number of vascular bundles are seen.
2.Vascular bundles are closed without cambium.
3.Xylem is arranged in shaped or’ V’ shaped
4. Ground tissue is not differentiated into cortex and pith.

Identification: So, the material is monocot stem.

A portion enlarged
 MONOCOT ROOT.T. S

Ground plan
Identification features of Root
1.Xylem is exarch.
2.Vascular bundles are radial.
3.Root hairs are present

Identification features of Monocot

1.More than six (polyarchy) xylem bundles are seen.
2.Pith is large.
3.Xylem is found round in cross section.

Identification
So, the material is monocot root.
A portion enlarged
 DICOT ROOT.T. S

Ground plan

Identification features of Root

1. Xylem is exarch.
2. Vascular bundles are radial.
3. Presence of root hairs.

Identification features of Dicot

1. The number of xylem and phloem are 3 to 5.
2. Pith is reduced.
3. Xylem is polygonal in section.
A portion enlarged
Identification
So, the material is dicot root.
 TAXONOMY

SOLANACEAE
Capsicum frutescens
Flower Flower.L.S

Floral Characters

- Actinomorphic Flower

- Bisexual Flower

K - Calyx - Five sepals, valvate and fused

C - Corolla -Five petals present, petals are fused, twisted aestivation.
A - Androecium- Five stamens, epipetalous. Floral Diagram
G – Gynoecium- Ovary is twisted, bicarpellary, axile placentation.

Floral Formula:

K (5) C (5) A 5 G(2)

 STUDY OF ANTHER STRUCTURE

Anther After Dehiscence

Identification Features
The given material is the cross section of an anther.
Anther or microsporangium consists of four wall layers.
The wall layers are epidermis, endothecium, middle layers and tapetum.
Several pollen grains are produced in anther.
 1

ECOLOGICAL INTERACTIONS

1. MUTUALISM – e.g; Lichen

Identification
The ecological interaction is mutualism (e.g; Lichen).

Identification Features
Mutualism is the interaction between two organisms in which
the two will get benefit.
Lichen is the association of fungi (mycobiont) and algae
(phycobiont).
Fungal partner gives moisture and shelter to algae; and algae
gives food to fungi.
 2

2. COMMENSALISM - e.g; Vanda

Identification
The ecological interaction is commensalism (e.g;Vanda).

Comments
Commensalism is the interaction between two organisms in
which one gets benefit and the other neither gets benefit or
harm.
Epiphytes are plants that grow on other plants only for support.
It does not take nutrients and water from the host.
 3

3. PARASITISM – e.g; Cuscuta

Identification
The ecological interaction is parasitism (e.g; Cuscuta).

Comments
Parasitism is the interaction between two organisms in which
one gets benefit and the other is harmed.
Cuscuta is a total stem parasite.
Food is absorbed from the host plant by using specialised root
called haustoria.
 4

3. PARASITISM – e.g; Loranthus

Identification
The ecological interaction is parasitism (e.g; Loranthus).

Comments
Parasitism is the interaction between two organisms in which
one gets benefit and the other is harmed.
Loranthus is a stem parasite.
Food is absorbed from the host plant by using specialised root
called haustoria.
 5

STUDY OF BIOTECHNOLOGICAL ASPECTS BY

OBSERVING PHOTOGRAPHS

1. CLONING VECTOR
Identification
The given material is cloning vector pBR 322

Comments
It is a plasmid vector.
It consists of ori site, recognition sites, antibiotic resistance sites and
rop site
Ori site is the site for starting the replication of plasmid.
ampR and tetR are antibiotic resistance sites.
EcoR I, BamH I, Sal I, etc. are restriction sites.
 6

2.BIOREACTOR

Identification
The given material is bioreactor.

Comments
Bioreactors are large vessels in which raw materials are biologically
converted into specific products like enzymes, hormones etc.
The volume of bioreactors may be 100 litres to 1000 litres.
Adequate temperature, pH, vitamin, oxygen etc. are providing for
proper growth of the culture.
 7

3. Bt. COTTON

Identification
The given material is Bt. Cotton

Comments
Genetically modified cotton crop is called Bt.cotton.
It is the Bt. gene introduced cotton plant.
Bt. indicates Bacillus thuringiensis.
This plant produce Bt. toxin and it kills certain insects that attacks
cotton plant.
 8

PHYSIOLOGY EXPERIMENTS
Experiment-1

STUDY OF STOMATA

Aim:
To study the distribution of stomata on upper and lower surface of leaves.

Materials required:
Fresh dicot leaves, needle, forceps, blade, brush, watch glass, microscope
etc.

Procedure:
Remove upper and lower epidermal peels of Hibiscus leaf into separate
watch glass containing water. From this take a separate square piece of each
peel. Each of this peel is mound in a separate slide and observe under
microscope. Count the number of stomata in upper and lower epidermis
through microscope.

Observation:
In dicot leaf Hibiscus the number of stomata present in upper epidermis is
less than the number on the lower epidermis.

Inference:
In dicot plant usually the number of stomata in lower side of the leaf is more
than the upper side (dorsiventral leaf).
 9

Experiment-2

STUDY OF PLANT PIGMENTS BY PAPER CHROMATOGRAPHY

Aim:
To separate the photosynthetic pigments by unidirectional paper chromatography.

Materials required:
Petroleum ether, acetone, Watman No. 1 filter paper, glass rod, measuring jar, split
cork, leaf extract etc.

Procedure: 2 ml of running solvent (petroleum ether and acetone in 9:1 ratio) is

taken in a large measuring jar. It is closed tightly with a sp-lit cork. A long strip of
Watman No;.l filter paper is taken and is cut at one end to get ‘V’ shaped tip. A
pencil line is drawn about 2cm above the tip and central point of it is marked as
origin. Leaf extract is prepared by grinding green leaves with acetone and filtering
it. The leaf extract is applied at the marked region of filter paper. A plenty of
pigments are applied on the filter paper and the spot should be less than 0.5 cm
diameter. The lower end of the filter paper is dipped in the solution and upper end
is fixed in the split cork in the jar.

Observation:
The pigments of the extract get separated because of their differential solubility in
a moving phase.
The lower most layer is chlorophyll ‘b’ (yellow green), then chlorophyll ‘a’ (blue
green), Xanthophyll (yellow orange), and Carotene (orange red) respectively
 10

Experiment-3
HYDRILLA EXPERIMENT

Aim:
To show evolution of oxygen during photosynthesis.

Materials required:
A few branches of Hydrilla plant, sodium bicarbonate (NAHCO3), a graduated
beaker, water, a glass funnel, a test tube, etc.

Procedure:
2 gm of sodium bicarbonate is dissolved in 10 ml of distilled water. 2 ml of this
water contains 0.4 grams of sodium bicarbonate. 100 ml of water is taken in a
graduated beaker. A few branched of Hydrilla are placed in a beaker. Cut the
bases of the branches of Hydrilla, tie them with thread and cover them with an
inverted glass funnel, in such a fashion that the cut ends of the plants are towards
the neck of the funnel. The funnel is completely under water. A test tube filled
with water is inverted over the tail of the funnel. The whole apparatus is kept in
sunlight and observed for some time.

Observation:
Small air bubbles come out continuously from the cut ends of Hydrilla and they
are collected at the top of the test tube by down ward displacement of water. On
testing this gas, it is found to be oxygen.

Inference:
From this experiment we can infer that the branches of Hydrilla carried on
photosynthesis and liberated oxygen.
 11

Experiment-4

YEAST FERMENTATION AND PRODUCTION OF ALCOHOL

Aim:
To demonstrate fermentation by yeast.

Materials required:
10% glucose solution, baker’s yeast, KOH, conical flask, test tube, glass
rod, lime water etc.

Procedure:
Take glucose solution in the conical flask and add a pinch of naker’s
yeast into it. Add a few drops of oil over the surface of the solution to
cut oxygen supply to yeast. One end of the delivery tube is inserted into
the conical flask above the solution through a one holed cork. The other
end of the delivery tube is inserted into the lime water taken in test tube.
Make the apparatus air tight by applying Vaseline. Keep the set up
undisturbed for 2 days.

Observation:
After two days the glucose solution will be fermented and alcohol smell
is coming out. Lime water turns milky white in colour.

Inference:
Yeast contains the enzyme zymase and it convert the glucose solution
into alcohol and CO2. So, glucose solution becomes alcoholic. The CO2
gas reach in the test tube through the delivery tube and react with lime
water to form white precipitate of calcium bicarbonate.
 12

End

Thank you
SUBRAHMANIAN. P.M
HSST BOTANY
GOVT. HSS NARIKKUNI