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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。
The hypocotyl callus of alfalfa (Medicago sativa L.) was treated with NaN_3 solution and continuously cultured on MS medium containing the lethal concentration of methionine for 6 months. 100 mmol / L methionine and differentiate into regenerated plants. The mutant cell line obtained was 7.2 times more resistant to methionine than control after six months of selective pressure release and showed cross-resistance to ethionine (3.3-fold that of the control) . The contents of methionine, lysine, threonine and isoleucine in the resistant cell lines and their regenerated plants both increased significantly compared with the control. SDS-PAGE and peroxidase isoenzyme bands of the resistant lines were significantly different from the control, and a new band appeared, indicating that the variation lines have produced changes in the gene product.