苜蓿抗甲硫氨酸变异体的筛选(英文)

SELECTION OF METHIONINE-RESISTANT VARIANT OF MEDICAGO SATIVA

紫花苜蓿(Medicago sativa L.)下胚轴愈伤组织用NaN_3溶液诱变处理后,在含有全致死浓度甲硫氨酸的MS培养基上进行了6个月的连续筛选培养,获得了能抗100mmol/L甲硫氨酸的变异细胞系,并分化成再生植株。所获变异细胞系在脱离选择压力6个月后,对甲硫氨酸的抗性仍比对照高7.2倍,并表现出对乙硫氨酸的交叉抗性(为对照抗性的3.3倍)。抗性细胞系及其再生植株的甲硫氨酸、赖氨酸、苏氨酸和异亮氨酸含量均比对照有大幅度增加。抗性系的SDS-PAGE电泳图谱及过氧化物酶同工酶谱带均与对照有显著不同,并出现了新带,表明变异系已经产生变化了的基因产物。

A variant cell line of Medicago saliva L. resistant to methionine (100 mmol/L) was selected .from calli treated with 1.5 mmol/L sodium azide. Plants were regenerated from this selected cell line. After growing for 6 months on the medium devoid of selection pressure, the variant cell line still showed high level of resistance, which was 7.2-fold higher than that of the wild type. This indicated the stability of the resistance expression. The variant cell line also expressed a 3.3-fold higher cross-resistance to ethionine, than that of the wild type. The contents of methionine, lysine, threonine and isoleucine in the variant calli were respectively 2.0, 1.43, 1.71 and 1.57 times higher than those in the wild type. However, in the variant regenerated plantlets they respectively were 2.0, 1.33, 1.22 and 1.22 times higher than those in the wild type regenerants. Four new protein bands, viz. 65 kD, 23 kD, 20 kD and 18 kD, appeared in the SDS-PAGE pattern of soluble proteins in the variant calli. In the peroxidase isoenzyme electrophoresis pattern, two new bands were also observed in the variant calli.