In Vitro Haploid Production by Anther Culture of Dong Du Mustard (Brassica juncea)

Abstract

     Androgenesis is a phenomenon in which microspores are made to bypass the sexual pathway and follow the sporophytic mode of development to generate new plants without the intervention of fertilization under in vitro conditions. Micorospore culture provides an ideal system, with a large, relatively uniform population of haploid cells, for use mutant selection, genetic transformation and in studies on the molecular mechanism of induction of androgenesis and embryogenesis. This paper aimed at establishing a protocol for anther microspore embryogenensis in traditional variety of Brassica juncea (Dong Du mustard). Five haploid lines was created from anther microspore culture. The results showed that, medium and plant growth regulator concentration had a pronounced effect on androgenic response in anther microspore cultures of Brassica juncea (Dong Du mustard). The medium B5 is the most suitable for morphogenesis of anther microspores culture. The highest percent of embryogenensis (2,33%) was obtained B5 medium supplemented with α-NAA 0,5 mg/l and Kinetin 1 mg/l. MS medium supplemented with IAA 0,5 mg/l, BA 0,5 mg/ l, 3% sugar and 8 g / l agar was capable of regenerating seedling from embryos. MS medium supplemented with 1,0 mg/l BA was optimal for shoot multiplication with a rate of 3.94 after four weeks. MS medium with 0,5 mg/l α-NAA was appropriate for root induction in terms of root number per shoot (6.54 root/shoot) and average length of root (3.68 cm) after three weeks.