In vivo and in vitro colchicine manipulations for enhancing doubled haploid production efficiency in bread wheat

The doubled haploidy breeding approach using chromosome elimination technique following wheat x maize system has provided the most suitable tool for doubled haploid production in bread wheat, but this has low success rate. Colchicines manipulations were done at in vivo and in vitro levels for enhancing the efficiency of doubled haploid production in bread wheat. For in vivo experiment colchicine concentrations ranging from 100 to 10000 ppm were injected with and without 2,4-D at first internode at different time intervals viz. 24, 48,72 and 96 hours after pollination. After 16-18 days embryos were excised and cultured on MS medium. For in vitro experiment the haploid embryos were rescued on MS medium containing colchicines at different concentrations ranging from 100 to 500 ppm and moved to colchicine free MS medium after different time intervals i.e. 24, 48, 72 and 96 hours. Control experiment was also conducted for both the methods in which colchicine treatment was not given neither in field nor in medium but at tillering stage by dipping the crowns of regenerated haploid plantlets in 0.1% colchicine solution supplemented with 1.5 ml/l dimethyl sulfoxide at 20 °C for 5 hours. Data on various parameters was analysed using three factorial RBD. The in vivo colchicine application was found to be most superior over standard method and in vitro method for enhancing doubled haploid production efficiency in bread wheat.