Page 1 : Anther Culture ie, , The culture of immature anthers into embryos is called, anther culture. \t was first done by Guha and Maheswari in, tobacco. Anther culture provides haploid plants. Now about, 250 species of dicot and moncot plants are regenerated from, anthers. Eg. wheat. maize, coconut, rubber, tobacco, etc., , In young anthers, the cells are diploid. The diploid, cells undergo meiosis to form haploid tetraspores. In the, culture medium the haploid tetraspores dedefifferentiated into, haploid parenchyma cells. Thus a mass of parenchyma cells, is produced in the anther. eo, , _ The cell mass becomes circular and forms a globular, embryo and. then heart-shaped embryo. Finally tarpedoshaped embryoid develops in the-culture medium. The, development of embryoid from the anther looks like-the, development of zygotic embryo. :, , Anther culture is very successful if microspores of, the anther have just completed meiotic division (Sunderland,, , 1979)., Procedure For Anther Culture —, 1. Unopened flower buds are collected from plants., , 2. The flower buds are surface sterilized with tween, 80 and mercuric chloride solution. Then they are washed with, - distilled water. :

Page 2 : ; ae Dissected stamens., Surface. , No e, ¥ sterilization J (f () ( ( | ( (|, nase, Unopen, , Flower buds é Acetocarmine, squash test, , Not : aid |, , , , , , , , suitable Suitable, for another, culture, Ig fad,, , , , , , , , Selected stamens., , , , , , , , , , , , Fig.20.12: Stages of anther culture., , 3. The anthers are taken from a flower buds carefully, , and kept ina petri disc. In such a Way anthers are taken from, all flowerbuds and put in separate petri discs.

Page 3 : 4, The anthers just completed first meiotic division., ected by acetocarmine squash test. :, '§, Suitable anthers are aseptically transferred toa liq, dor solid nutrient meditim containing glutamine. L-serine, ul ah : ;, and inositol. ;, ¢, , 6. The culture plates are incubated at 25°C for about, {5 days. During the incubation anthers grow into embryoids., , 7. The embryoids are transferred toa rooting medium, in the culture plates 3000 lux illumination is given to the, culture. The embryoids become plantlets within 4-5 weeks., , _8. The well developed haploid plantlets are planted in, pots in a green house for proper acclimatization. , Uses of Anther Culture, , 1. Anther culture is used to produce haploid plantlets., They have a single set of chromosomes in the cells. So the, recessive genes also express their traits in the plants. Fhe, recessive characters can easily be identified in haploid plants., , 2. Haploids have reduced number of genes so the possibilities of metabolic complexities are minimum. Hence, they are used in the study of metabolic pathways., , 3. Anther culture is of much tse in invitro mutagenesis, for crop improvement., , 4. Haploid plants do not produce fertile gametes. This, is due to abnormalities during gametogenesis. especially in, , meiosis. The chromosome number of haploids is doubled by, , colchicine treatment. The haploid embryoids are immersed, , In colchicine for two days and grown into plants in vitro. The, resulting plants are diploid and fertile., , ____ Application of colchicine paste to axillary buds of haploid plants is found to be effective in chromosomal doubling., , Here. branches of the haploid plants become diploid and, fertile, fates i, , re Se!, , ¢Q