苜蓿培养细胞抗羟脯氨酸变异体的筛选和特性分析  

SELECTION AND CHARACTERISTICS ANALYSIS OF L HYDROXYPROLINE RESISTANT VARIANT FROM MEDICAGO SATIVA L. CALLUS CULTURES

作　　者：王瑛[1,2] 牛炳韬[1,2] 贾敬芬 

机构地区：[1]兰州大学生物系 [2]河北大学生物系

出　　处：《应用与环境生物学报》1998年第4期340-344,共5页Chinese Journal of Applied and Environmental Biology

基　　金：国家教委博士点基金

摘　　要：以返地卫星搭载的苜蓿（ＭｅｄｉｃａｇｏｓａｔｉｖａＬ．）种子为实验材料，取其无菌苗下胚轴切段诱导愈伤组织，该愈伤组织经正筛选法，获得抗脯氨酸类似物Ｌ羟脯氨酸（Ｈｙｐ）的变异细胞系（Ｈｙｐｒ）．Ｈｙｐｒ脱离选择压力３ｍｏ后，对Ｈｙｐ的抗性仍比对照强，ＦＷ细胞内游离脯氨酸ｗ＝０．７２ｍｇ／ｇ，是ＣＫ的３．４３倍，同时还具有对ＮａＣｌ和ＰＥＧ的交叉抗性．和ＣＫ相比，Ｈｙｐｒ愈伤组织可溶性蛋白质ＳＤＳＰＡＧＥ图谱出现两条新多肽（Ｍｒ≈５６×１０３、３２×１０３）；其过氧化物酶同工酶谱中酶活性明显高于对照，并出现１条新带，酯酶同工酶酶谱中亦出现３条新带．这些特性分析表明，该变异细胞系对Ｈｙｐ的抗性是稳定的．By positive selection method in vitro , L hydroxyproline resistant variant (Hyp r) was obtained from hypocotyl derived calli of Medicago sativa (Alfalfa) seedlings, whose seeds were tested in a satellite having returned in 1994. After 3 months of subculturing without selection pressure, Hyp r maintained the resistance to L Hyp and its free proline mass fraction ( w ) was 0.72 mg/g, which was 3.43 folds of that in control. Hyp r also showed high cross resistance to NaCl and PEG. Compared to the control, two new bands appeared in SDS PAGE patterns and a new band in the peroxidase isoenzyme patterns, as well as three new bands in the esterase isoenzyme patterns from the soluble proteins of Hyp r calli. The results indicated that Hyp r had produced the proteins encoded by changed genes and its resistance to L Hyp was stable.

关 键 词：苜蓿 愈伤组织 抗羟脯氨酸变异系 生理生化特性 

分 类 号：S551.703.5[农业科学—作物学] Q944.6[生物学—植物学]