激活标签转化结球甘蓝的研究  

Transformation of Cabbage(Brassica oleracea L.) Using Activation Tagging Plasmid

作　　者：王凤华[1] 陈双臣[1] 李光远[1] 王丽敏[1] 

机构地区：[1]河南科技大学林学院,河南洛阳471003

出　　处：《西北植物学报》2009年第5期905-909,共5页Acta Botanica Boreali-Occidentalia Sinica

基　　金：河南省自然科学基金(2007210004);河南科技大学人才科学研究基金(05-126)

摘　　要：以夏光甘蓝下胚轴为材料,采用含激活标签pSKI015的农杆菌GV3101进行遗传转化研究,对预培养时间、侵染时间和共培养时间等转化条件进行优化.结果表明,预培养2d,侵染6min,共培养1d是遗传转化的较优组合.采用GV3101转化下胚轴结合glufosinate筛选,获得了2株表型变异植株:一株叶片中心绿色,边缘黄化;另一株叶片卷曲.PCR检测显示这2株植株均含有pSKI015增强子序列,Southern blot结果显示这2株植株具有明显的杂交信号,说明这2株植株为激活标签突变体.Genetic transformation for cabbage was studied using hypocotyls of ＇Xiaguang＇ variety and Agrobacteriurn turnefacierns GV3101 （with activation tagging plasmid pSKI015）. The transformation factors of cabbage,including pre-culture time,invading time and co-culture time were optimized. The results showed that pre-culture for two days, invading for six minutes and co-culture for one day was the best treatment for cabbage transformation. Two plantlets were obtained invading hypocotyls of cabbage with GV3101 and screening on medium added with 1. 5 mg/L glufosinate. Of those two plantlets, one with yellow-edge and green central leaves, the other with roll leaves. Results of PCR amplification and southern blot showed that enhancer of pSKI015 had been inserted into genome of these two plantlets and thus they were activation tagging mutants of cabbage. Activation tagging transformation system for cabbage was established in this experiment, which was the basis for construction library of activation tagging mutants.

关 键 词：甘蓝 遗传转化 激活标签 突变体 

分 类 号：Q813.2[生物学—生物工程]