Professional Documents
Culture Documents
Content
Totipotency Historical background Requirements for cell and Tissue Cultures A tissues culture laboratory Nutrient media Inorganic chemicals Growth hormones Organic constitutents Vitamins Amino acids Culture of plant materials Explant culture Callus formation and its culture Organogenesis Root culture Shoot culture and micropropagation Cell culture Benefits from cell culture Somatic embryogenesis Somaclonal variation Protoplast culture Isolation Regeneration Protoplast fusion and somatic hybridization Fusion products Method of somatic hybridization Anther and pollen Culture Culturing techniques In vitro androgenesis (direct and indirect androgenesis) Mentor pollen technology Embryo culture Embryo rescue Protoplast fusion in fungi
Culturing techniques
Methods of anther culture are shown in Fig. 8.9. Anthers are superficially sterilized and washed with double distilled sterile water. They are excised from the flower buds and their proper developmental stages are determined under microscope. On confirmation of a stage, (a) the anthers are directly transferred on nutrient agar or liquid medium where induction of embryogenesis occurs, or (b) the pollen grains are aseptically removed from the anthers and cultured on liquid medium (Reinert and Bajaj, 1977).
Fig. 8.9. Methods of anther culture, and production of haploid and diploid plants in vitro.
This diagram shows the various stages of anther and isolated pollen culture. The stages of anther culture from anther to haploid plantlet can be described as follows: a) an unopened flower bud, 1b) anthers, 1c) the anthers in culture, 1d) and 1e) proliferating anther, 1f) haploid callus, 1g) differentiating callus, h) haploid plantlet. Isolated pollen culture is as follows: a) an unopened flower bud, 3b) isolated pollen from a cultured anther, 3c) pollen culture, 3d) multinucleate pollen, 3e) and 3f) pollen embryo. Homozygous plants can be obtained by treating the haploid plantlets with colchicine.