CoreExperiments 43

EXPERIMENT 6.1
To
prepare temporary acetocarmine stained mount of onion root tip to study various
A I M

stages ofmitosis.

R E Q U I R E M E N T S

Onion bulb,
conical flasks/glass bottles, corked vial/tube,
petridishes, scissors, forceps, needles,
l alcohol, acetic acid, hydrochloric acid, acetocarmine, distilled water, spirit lamp, microscope,

slides,
coverslips blotting paper et.

PROCEDURE

of a sharp
Take a medium sized bulb of onion and trim off the old roots from its base by means
blade.
its base touching the water.
Place the onion o n a conical flask/glass bottle full of water, with
Keep it for a week to grow the roots.
acid
Cut 5 mm off the of roots and put them into a vial containinga mixture of 1:3 acetic
tips
root tips should be
and methanol. Keep for one hour. This process is called fixation. (Cutting of
summer and between 9.30 a.m.
to
done in the morning between 7.00 a.m. to 8.00 a.m. during the
11.30 a.m. during the winter).
acid for
4 Remove 2 or 3 root tips and hydrolyse them by warming to 60°C in 1 Nhydrochloric
15 minutes.

Onion bulb Onion

Roots

Bottle
EVNN Onion root

Water Beaker
- Water

6.1. Method of growing onion root tips.

Fig.
. in water.
Kemove the tips and wash them thoroughly
root
one hydrolysed root tip
in a drop and place a coverslip
lace a drop of acetocarmine on a slide. Put
on the root.
with the blunt end of a pencil or needle until
Gent
Ently squash the root by tapping the coverslip
the
e u s separate and spread out into a very thin layer.

M
E Sure that there are n o air bubbles under
the coverslip.
. few seconds.
.
Gently warm the slide over a flame for a

the
first under the low power of the microscope to locate the dividing cells. Examine
erve of the microscope.
rent
stages of mitosis under the high power
 Manual in Biology-XII
44 Comprehonsive Laboratory

OBSERVATIONS are seen

scattered. Under
Unaer loW power of the microscope, rectangular cells
with pink nucleus
(Figs. 6.2 and
0.)

hgh power of themicroscope following stages become distinct:

Maturation zone-

Prophase
Zone of elongation
Metaphase

Meristematic zone
Telophase

Anaphase

Interphase

6.2. Different stages of mitosis in

the onion root tip.
Fig.

Interphase cell divisions.

successive
) It is a non-dividing phase of the cell cycle between two
(i1) Chromatin fibres appear in the form of a network within the nucleus.
distinct.
(iii) Nuclear envelope and nucleolus are

Prophase
called chromosomes.
() Chromatin material shortens and condenses into thread like structures
at a point called centromere.
(ti) Each chromosome consists of two chromatids, jointed
(ii) Nuclear membrane and nucleolus start disintegration and disappear at the end of prophase.

3- Metaphasse
() A bipolar spindle develops in the cell. Chromosomes become thick and two chromatids of
each chromosome become clear.
(ti) Chromosomes become arranged at the equator of the spindle.
(it) Each chromosome get attached to the spindle fibres at its centromere.
4. Anaphase
(1) The two sister chromatids of each chromosome separate from the centromere and move
towards the opposite poles.
Gi) The daughter chromosomes (separated chromatids) appear V, J L and I shapes, depending
upon the position of centromere.
5. Telophase
() The spindle disappears and the daughter chromosomes uncoil to form
chromatin fibres at
the two poles.
(i) Nuclear membrane and nucleolus
reappears and e
two daughter nuclei appear at
oppos1
poles.
it Cytokinesis occurs by cell plate formation between the two
daughter nuclei.
Core Experiments

45

Nuclear
membrane

Chromatin
fibres

Nucleolus

Cell membrane

Interphase stage
Cell wall Nuclear
Nuclear membrane
membrane
Disappearing
Nucleoluss nucleolus

Chromosomes Chromosomes
Cell wall

Early prophase
Late prophase

Spindle Daughter
fibres chromosomes

Chromatids
-Spindle fibres

Early anaphase
Metaphase stage
Daughter cells
Cell wall
Daughter nuclei
Chromosomes
(Chromatids) - Cell plate

Nuclear
Spindle fibres membrane

Late anaphase Telophase stage

Fig. 6.3. Various stages of mitosis in onion root tip cells.

PRECAUTIoNS
ne base of the onion bulb should be in contact of water while growing the roots.
2.
3. Ot tips should be fixed in the morning between 8 to 10 A.M.
The slide warmed gently much above the flame of the spirit lamp.
should be