摘要
用ＮａＮ３ 诱变过的红豆草（Ｏｎｏｂｒｙｃｈｉｓｖｉｃｉａｅｆｏｌｉａ Ｓｃｏｐ．）愈伤组织筛选得到了能抗８０ ｍ ｍ ｏｌ／Ｌ甲硫氨酸的变异细胞系——ＯＮｍ ｅｔｒ，并得到了再生植株。ＯＮｍ ｅｔｒ 脱离选择压力６ 个月后，其抗性仍比野生型的高５．６ 倍。同时还表现出对乙硫氨酸的交叉抗性，其抗性是野生型的６．５ 倍， 表明该抗性系抗性表达稳定。ＯＮｍ ｅｔｒ 愈伤组织中甲硫氨酸、赖氨酸、苏氨酸含量分别是野生型的４．００、１．０９、１．５０ 倍。ＯＮｍ ｅｔｒ再生植株中甲硫氨酸、赖氨酸、苏氨酸、异亮氨酸含量分别为野生型的２．０、３．５、３．５、２．５ 倍。在ＯＮｍ ｅｔｒ 愈伤组织可溶性蛋白ＳＤＳ－ＰＡＧＥ图谱中，出现了两条新多肽（３０ ｋＤ、２６ ｋＤ）。ＯＮｍ ｅｔｒ 细胞系过氧化物酶同工酶谱中亦出现了两条新酶带。
The variant cell line of Onobrychis viciaefolia Scop.(ONmet r)resistant to 80 mmol/L methionine was isolated from calli which was treated with NaN 3. This ONmet r cell line was induced to regenerate plantlets. After growing for 6 months on a medium without selection pressure,the ONmet r cell line was still highly resistant to methionine being 5.6 fold higher than that of the wild type. The variant cell line also expressed high level of cross resistance to ethionine which was 6.5 fold higher than that of the wild type. The contents of total methionine(Met)、lysine(Lys)、threonine(Thr) in ONmet r calli were 4.00、1.09、1.50 fold respectively higher than those of the wild type. The contents of total Met、Lys、Thr、Ile(isoleucine) in ONmet r regenerants were 2.0、3.5、3.5、2.5 fold respectively higher than those of the wild type. Two new bands appeared in SDS PAGE profile as well as in the superoxidase isoenzymes electrophoresis pattern of the soluble proteins of ONmet r calli,thus indicated that the variant had carried the products of the changed genes.
基金
国家教委博士点基金
关键词
红豆草
甲硫氨酸
变异体
筛选
Onobrychis viciaefolia
Methionine
Variant selection
Regenerated plantlet
