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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。
In this study, Agrobacterium-mediated transformation of maize zygotes was established. Agrobacterium tumefaciens-mediated transformation method was used, the standard binary vector PTF102 containing the bar gene and the vector p3300-P containing the bar gene and the bivalent insect-resistant genes Cry1A (a) or Cry1A (c) and PTA (Pinellia ternata) Bt-pta was transformed into maize inbred line Ji 8902, Dan 340, Ji 4112, Ji 853, Tie 7922 and PA91. Transformed seeds were directly obtained from recipient plants. The transgenic plants and progeny were obtained by PPT screening and PCR identification. The seed setting rate, transformation rate and genetic inheritance of three batches of transformation were experimentally analyzed in 2002, 2003 and 2004: the average seed set rate of the infected ears was 39% and the frequency of zygotic transformation was over 1% Transgenic can be inherited. Agrobacterium-mediated maize zygotic transformation method can be repeated success, indicating that we have successfully established a new tissue culture-free corn gene technology system.