茎用芥菜细胞质雄性不育系子叶原生质体培养和高效植株再生(英文)

Efficient Plant Regeneration from Cotyledon-derived Protoplasts of Cytoplasmic Male-sterile Tuber Mustard (Brassica juncea Coss.var.tumida Tsen et Lee)

利用茎用芥菜细胞质雄性不育系原生质体培养获得了再生植株 ,并研究了影响原生质体培养的因素。结果表明 ,子叶是茎用芥菜原生质体培养最佳的外植体 ,1 0d苗龄的子叶原生质体在改良MS培养基上培养 3d后发生第 1次细胞分裂 ,6d后发生第 2次分裂 ,3周后形成细胞团 ,5周后形成肉眼可见的小愈伤。培养基中缺少NAA或 2 ,4 D都会降低愈伤组织的再生能力。在含一定浓度的NAA( 0 .2 5mg/L)和 2 ,4 D( 0 .2 5mg/L)培养基上诱导的愈伤组织质地致密且有光泽 ,芽的分化能力高 ;在MS +BA 1mg/L +NAA 0 .2mg/L的培养基上芽的分化频率高达近 2 9% ,再生芽在 1 /2MS +NAA 0 .1mg/L培养基上生根 。

Plants of cytoplasmic male sterile (CMS) tuber mustard were regenerated from cotyledon derived protoplasts. The factors affecting protoplast culture and plant regeneration were studied. Protoplasts isolated from 10 day old seedlings of CMS tuber mustard showed cell division upon culture on modified MS medium, and microcalli were produced at a high plating efficiency of nearly 16% after a 5 week culture. Various combinations and concentrations of growth regulators were added to the medium for the proliferation of microcalli, and to obtain regenerants from the calli. The results showed that NAA or 2,4 D was necessary for callus proliferation. The absence of NAA and 2,4 D resulted in production of the callus with low regenerative potential, and in the presence of NAA ( 0.25 mg/L) and 2,4 D(0.25 mg/L), compact and bright calli were produced with high reproductive potential. Regenerated shoots were obtained at a frequency of nearly 29% when the compact and bright calli were selected and transferred at an early stage to the medium containing BA (1 mg/L) and NAA( 0.2 mg/L). All the regenerated shoots were rooted on 1/2 MS medium with NAA 0.1 mg/L added.