摘要
选择青海高原海拔 40 0 0m高山上生长的唐古特红景天为实验材料 ,以叶片为外植体 ,在MS +BA2 +NAA0 2 固体培养基上诱导出黄绿色、松脆愈伤组织 .愈伤组织细胞在同样成分的液体培养基中培养获得成功 .在悬浮培养液中可检测到分泌蛋白的存在 .经SDS 聚丙烯酰胺凝胶电泳分析表明 ,低温锻炼或脱落酸 (ABA)诱导后 ,细胞分泌蛋白的多肽谱带数增加 .与此相对应的是 ,细胞的抗冻能力也明显提高 .PAS染色揭示多肽中均含有糖基 .通过测定热滞值 。
The culture of callus and cell suspension induced from leaves of Rhodiola algida Var.tangutica collected from Qinghai plateau(altitude: 4 000 m ),on solid and in liquid culture medium(MS+BA 2+NAA 0 2 )respectivelly,was successful.After cold (4～5℃)acclimation or ABA treatment for 10 or 14 days, the freezing resistance of suspension cells was increased. Proteins from the liquid suspension cell culture medium exhibited antifreeze activity. Treatment of the antifreeze proteins(AFPs) with boiling or trypsin caused inactivation,proving the protein characteristics of AFPs.Periodate and borate inhibited the antifreeze activity of AFPs,suggesting the presence of carbohydrate. Of the four treatments(25℃;25℃+ABA;5℃;5℃+ABA),2～3 polypeptides(29～50 ku) could be detected from the one dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS PAGE). PAS(periodic acid schifts reagent) reaction of SDS PAGE showed the presence of carbohydrate in the polypeptides.
出处
《生物化学与生物物理进展》
SCIE
CAS
CSCD
北大核心
2000年第5期555-559,共5页
Progress In Biochemistry and Biophysics
基金
国家自然科学基金!(3980 0 118)
霍英东青年教师基金!(710 30 )
关键词
高山植物
唐古特红景天
悬浮培养细胞
抗冻蛋白
alpine plant, Rhodiola algida Var. tangutica, suspension cell culture, antifreeze proteins
