甘蓝子叶原生质体培养与植株再生的研究

The Culture of Cotyledon Protoplasts and Regeneration of Plantlets of Brassica oleracea

将0.5％纤维素酶Rs+0.1％果胶酶Y-23酶溶液振荡处理3h,可得到最高的细胞分裂率,但原生质体分离数量较低。0.5％纤维素酶R-10+0.05％离析酶R-10+0.05％果胶酶Y-23酶溶液,25℃下处理14h,原生质体分离数量及细胞分裂率均较高。1/2MS培养基添加NAA3mg/L+BA0.5mg/L+2,4-D 0.1mg/L的液体培养基有利于细胞团形成及愈伤组织生长。本试验获得了完整的再生植株,芽分化率达20％左右。

Through the treatment of application of 0.5% cellulase Rs+0.1% pectolyase Y-23 by shaking for 3h, the highest cell division efficiency was obtained, but the number of protoplast isolation was relatively low. Also, through the treatments using enzyme mixture of 0.5% cellulase R-10+0.05% macerozyme R-10+0.005% pectolyase Y-23 incubated at 25℃: for 14h, more protoplasts were isolated and a high efficiency of cell division was gained. The liquid medium of 1 / 2 MS containing 3 mg/ L NAA, 0.5 mg/ L BA and 0.1 mg/ L 2,4-D was favourable to the formation of cellular aggreate and the callus growth. Complete plantlets regenerated were obtained, and the bud differentiation rate can reach 20% or so in this experiment.