甘蓝下胚轴原生质体再生植株

Plant Regeneration from Hypocotyl Protoplasts of Brassica oleracea

经纯化后,甘蓝下胚轴原生质体的产量为1.5×10~6g^(-1)(FW),采用液体浅层培养的方法进行培养。2～3d后,发生第一次分裂,第10天,统计分裂频率为61％,5周内形成大量的细胞团和小愈伤组织,统计植板率为1.1％,把小愈伤组织转到与原生质体培养基相同激素的MS固体培养基上增殖。当愈伤组织长到3～5mm大小时,接到分化培养基上,芽分化率为46.7％,分化出来的芽长到3～4cm长时,从基部切下,插入生根培养基,两星期左右即可长成完整植株。

The yield of purified protoplasts isolated from hypocotyl of Brassica olear-acea was 1. 5 X 106g -1 (FW)and they were cultured in a K8p liquid medium supplemented with 0. 2 mg/L 2, 4-D, 1mg/L NAA and 0. 5mg/L BA. The protoplasts started to divide after 2~3 days of culture, and the division frequency was 61% after 10 days. The cell clones and microcalli could be obtained in 5 weeks, then transferred to the proliferation MS medium supplemented with the same hormones as the K8p liquid medium. The plating frequency was about 1.1%. Calli of 3~5mm in diameter were transferred on MS differentiation medium, the frequency of shoot but formation was about 46. 7%. Whole plants were obtained upon transferring 3~4cm shoots to 1/2 MS medium with 0. 5mg/L IBA.