农杆菌介导的玉米合子基因转化  

Zygote Transformation Using Agrobacterium-mediated Method for Maize

作　　者：刘德璞[1] 袁英[1] 郝文媛[1] 王景会[1] 李晓辉[1] 孔祥梅[1] 姜志磊[1] 孙传波[1] 曲文利[1] 

机构地区：[1]吉林省农业科学院生物技术研究中心,长春130033

出　　处：《分子植物育种》2008年第5期874-880,共7页Molecular Plant Breeding

基　　金：国家转基因植物中试及产业化基地(吉林)专项(JY03-17-1)资助

摘　　要：本研究建立了农杆菌介导的玉米合子转化方法。用农杆菌介导合子转化方法,以含有bar基因的标准双元载体PTF102和含有bar基因和双价抗虫基因Cry1A(a)或Cry1A(c)、PTA(半夏凝集素)的载体p3300-Bt-pta转化玉米自交系吉8902、丹340、吉4112、吉853、铁7922及PA91,直接从受体植株得到转化种子,用除草剂PPT筛选和PCR鉴定,获得转基因植株及后代。实验分析了2002年、2003年和2004年的3批转化操作的结实率、转化率及转基因的遗传情况:经农杆菌侵染的雌穗平均结实率为39%,合子转化频率达1%以上,转基因可以遗传下去。农杆菌介导玉米合子转化方法可以重复获得成功,表明我们成功建立起一个新的不依赖组织培养的玉米转基因技术体系。A gene transfer method of Agrobacterium-mediated maize zygote transformation have been established. Using the method, standard binary （non-super binary） vector PTF 102 containing bar selectable marker gene and p3300-Bt-pta containing bar selectable marker gene and two interest genes CrylA （a） or CrylA （c） and PTA were transferred into 6 maize inbred lines of Ji8902, Dan340, Ji4112, Ji853, Tie7922, PA91. The transformed seeds were obtained direct from receptor plants and transgenic plants were selected from seeding with Basta herbicide, which were further identified to be genetic by PCR analysis. The analysis of the rate of seed setting of infected ears, transformation frequency and genetics oftransgene, from 3 year tests of 2002, 2003, 2004 showed that rate of seed setting is 39% and transformation frequency is up to more than 1%, that the method can be repeated successfully and transferred gene can heredity. Further a novel system of tissue transformation for maize have developed by us.

关 键 词：玉米 合子 农杆菌介导法 基因转化 

分 类 号：S513[农业科学—作物学]