Plant Regeneration from in vitro Cultured Anthers of Black Mustard (Brassica nigra Koch)

Abstract

AbstractAnthers of Brassica nigra, excised from fresh as well as cold‐pretreated (3 days at 3 ± 2°C) buds cultivated on modified B5 medium (Gamborg et al. 1968) containing sucrose level varying from 2 % to 10 %, along with 1O−6M BAP (benzylaminopurine) and 9 × 10−6M 2,4‐D (2,4‐dichlorophenoxyacetic acid), developed calli and/or embryos. The latter response was observed only in anthers reared on media containing 6 % or higher levels of sucrose. On media containing two or four per cent sucrose, the anthers produced calli, exclusively. The growth of embryos was inhibited or else they started callusing if left on the media containing higher levels of sucrose. However, on transfer to MS medium (Murashige and Skoog 1962), containing 2 % sucrose, embryos started callusing and subsequently a few secondary embryos differentiated. Such embryos were sub‐cultured on MS + 5 × 10−6M BAP + 2 % sucrose, wherein numerous shoots developed from embryos. The shoots were rooted by transferring to a medium containing 5 × 10−6M NAA (naphthalene acetic acid). Within two months of culture, some of these plants started flowering in vitro.