紫花苜蓿基因转化组织再生过程中褐化因素的分析  

Analysis and Solutions on Browning Factors in the Process of Genetic Transformation and Tissue Regeneration of Alfalfa(Medicago sativa L.)

作　　者：李云霞[1] 马晖玲[1] 成慧[2] 

机构地区：[1]甘肃农业大学草业学院/草业生态系统教育部重点实验室,甘肃兰州730070 [2]兰州大学草地农业科技学院,甘肃兰州730000

出　　处：《中国草地学报》2010年第4期69-74,共6页Chinese Journal of Grassland

基　　金：甘肃省农业生物技术研究与应用开发项目"紫花苜蓿抗病转基因植株的选育"(GNSW-2007-03);国家科技支撑计划"奶牛专项"子课题1"优质高产抗逆苜蓿和饲料作物新品种选育"子专题(2006BAD04A04-01-08)

摘　　要：以新疆大叶苜蓿、甘农3号紫花苜蓿和陇东苜蓿的下胚轴为外植体,以农杆菌介导法进行了抗旱基因的转化,针对转化组织在再生过程中出现的褐化现象,分析了转化后的外植体愈伤组织于冬夏两季在诱导、分化及再生过程中的褐化因素,并提出了解决褐化的办法。结果显示:在非季节性影响因素中,2,4-D的作用时间以15～20d为宜,超过30d褐化程度加深;蔗糖浓度在诱导、分化及再生3个阶段分别以30g/L、20g/L、10g/L为宜;基因型也影响褐化指数的高低。在季节性影响因素中,培养温度冬季以恒温25℃为宜,夏季以白天23℃+黑夜21℃为宜;光照以2000～3000lx为宜;夏季还要降低激素浓度或间断性去除激素作用来降低褐化。Hypocotyls of three varieties of alfalfa（Xinjiang Daye,Gannong No.3 and Longdong） were used as explants.Drought-resistant gene was transformed with agro bacteriummediated method.For the browning phenomenon occurred in the regeneration process,factors of browning which appeared in the induction,differentiation and regeneration of transformed callus of the explants in summer and winter were analyzed and some solutions were suggested.The study showed that,in the non-seasonal factors,the appropriate action time of 2,4-D was 15 to 20 days.A deeper browning level would be gotten above more than 30 days.Appropriate sucrose concentration of each of the three stages（induction,differentiation and regeneration） was 30g/L,20g/L,10g/L.Genotype also affected the level of browning.In the seasonal factors,appropriate incubation temperature in winter was 25℃ of constant temperature and in summer was 23℃ during the night and 21℃ during the day.Suitable light was about 2000～3000lux.Browning could be reduced by reducing the hormone concentration or intermittently removing hormones in summer.

关 键 词：紫花苜蓿 基因转化 再生 褐化 

分 类 号：Q943.1[生物学—植物学]