eee TS 7PQUIREMENTS Jglass bottles, corked vial/tube, petridishes, scissors, forceps, acid, hydrochloric acid, acetocarmine, distilled water, spirit ‘slips, blotting paper etc. Onion bulbs, conical fl needles, methyl alcohol, aceti lamp, microscope, slides, cove ——anDURE PROCEDURE —_— 1, Take a medium sized bulb of onion and trim off the old roots from its base by means of a sharp blade. 2, Place the onion on a conical flask/glass bottle full of w: water. Keep it for a week to grow the roots. 3. Cut 5 mm off the tips of roots and put them into a vial cont acetic acid and methanol. Keep for one hour. This process is ¢ root tips should be done in the morning between 7.00 a.m. to 8.00 a.m. during th and between 9.30 a.m. to 11.30 a.m. during the winter). 4, Remove 2 or 3 root tips and hydrolyse them by warming to 61 for 15 minutes. ater, with its base touching the aining a mixture of 1: 3 alled fixation. (Cutting of e summer 0°C in 1 N hydrochloric acid Onion Onion root Beaker Water Fig. 6.1. Method of grewing onion root tips em thoroughly in water. 5. Remove the root tips and wash th drolysed root tip in a drop and place 6. Place a drop of acetocarmine on a slide. Put one hy ‘a coverslip on the root. 7. Gently squash the root by tapping tl until the cells separate and spread out into a very thin | Make sure that there are no air bubbles under the coverslip. 8. Gently warm the slide over a flame for @ few seconds. 9. Observe first under the low power of the microscope to locate the dividing cells. Exam- ine the different stages of mitosis under the high power of the microscope. he coverslip with the blunt end of a pencil or needle yer. OBSERVATIONS: stangular cells with pink nucleus are seen e microscope, Tec following stages become distinct : (Figs. 6.2 Under low power of th f the microscope scattered. Under high power 0 and 6.3)Interphase (i) It is a non-dividing phase of the cell cycle between two successive cell divisions. (ii) Chromatin fibres appear in the form of a network within the nucleus, (iii) Nuclear envelope and nucleolus are distinct. Prophase (i) Chromatin material shortens and condenses into thread like structures cally chromosomes, (ii) Each chromosome consists of two chromatids, jointed at a point called (iii) Nuclear membrane and nucleolus start disintegration and disappear at the end prophase. Metaphase (i) A bipolar spindle de of each chromosome become clear. (ii) Chromosomes become arranged at the equator of the spindle, (iii) Each chromosome get attached to the spindle fibres at its centromere, Anaphase (i) The two sister chromatids of cach chromosome separate from the centromere ao move towards the opposite poles, (ji) The daughter chromosomes (separated chromatids) appear V, J, L and I shay depending upon the position of centromere, Telophase (i) The spindle disappears and the daughter chromosomes uncoil to form fibres at the two poles, ; (jj) Nuclear membrane and nucleolus reappears and two daughter nuclei appe opposite poles. (ii) Cytokinesis occurs by cell plate formation between the two daughter nue ntromere ops in the cell. Chromosomes become thick and two chromate chro®”PRECAUTIONS The base of the onion bulb should be in contact of water while growing the roots. Root tips should be fixed in the morning between 8 to 10 A.M. The shde should be warmed gently much above the flame of the spirit lamp. adInterphase stage | Cell wall Nuclear membrane |~—— Nucleolus Chromosomes Spindle fibres |_—— Chromatids Cell wall Chromosomes (Chromatids) Spindle fibres Late anaphase Nuclear membrane Chromatin fibres Nucle — Cell membrane Nuclear _— membrane |___ Disappearing nucleolus Chromosomes |—— Cell wall Daughter —~ chromosomes — Spindle fibres — Daughter cells Daughter nuclei — Cell plate Nuclear membrane Telophase stage