将GNA基因导入宁夏枸杞及其表达的研究  

A Study of the Introduction and Expression of GNA Gene in Lycium bararum L.

作　　者：曲玲[1] 曹有龙[2] 侯玉霞[1] 吴家和[3] 罗青[2] 田颖川[3] 

机构地区：[1]中国农业大学理学院,北京100094 [2]宁夏农林科学院,宁夏银川750002 [3]中国科学院微生物研究所,北京100080

出　　处：《西北林学院学报》2007年第3期88-91,共4页Journal of Northwest Forestry University

基　　金：宁夏回族自治区自然科学基金项目(B1006)

摘　　要：为了解决枸杞蚜虫侵害的难题,以宁夏枸杞主栽品种“宁杞1号”叶片为外植体,通过农杆菌介导的转化方法,将改造后的雪花莲凝集素GNA基因转入枸杞,以抗性愈伤组织诱导率作为转化效率的指标,对影响转化效率的因素进行了研究,初步建立了转化频率较高的转化系统。对获得的70个转基因株系进行PCR检测,阳性率为60%。运用Western blot杂交检测了2个转化子叶片和果实中GNA的表达,结果表明,其中一个转化株系在维管组织成分较高的叶片、青果中得到了GNA的表达,而在维管束成分极少的红果中未检测到GNA蛋白的表达。A modified Galanthus nivalis agglutinin （GNA） gene was introduced into the genome of main cultivar ＇Ningqi No. 1＇ of Lycium bararum in vitro with explants of leaf disks mediated by Agrobacterium tumefaciens. Several factors affecting transformation efficiencies were studied based on induction rate of resistant callus. An effective system for genetic transformation of Lycium bararum was set up, and 70 kanamycin-resistant transformants were obtained. Results of PCR indicated that 60 % kana resistant plants could be detected the gna gene. Western blot analysis of leaf and fruit proteins from two PCR positive and aphid resistant transformants was carried out. In one of transformants, the GNA protein was detected in leaves and green fruits where the vascular tissues occupies a relatively larger part of the organs, but in red fruits where the vascular tissues occupy few parts of the organs the GNA protein wasn＇t detected.

关 键 词：宁夏枸杞 雪花莲凝集素 CoYMV启动子 农杆菌介导遗传转化 Westernblot杂交 

分 类 号：S793.904[农业科学—林木遗传育种]