Development of Marker-Free Transgenic Cry1Ab Rice with Lepidopteran Pest Resistance by Agrobacterium Mixture-Mediated Co-transformation

作     者：QI Yong-bin YE Sheng-hai Lu Yan-ting JIN Qing-sheng ZHANG Xiao-ming 

作者机构：Institute of Crop and Nuclear Technology Utilization Zhejiang Academy of Agricultural Sciences Hangzhou 310021 China Biotechnology Institute Zhejiang University Hangzhou 310029 China 

出 版 物：《Rice science》 (水稻科学（英文版）)

年 卷 期：2009年第16卷第3期

页      面：181-186页

学科分类：09[农学] 0904[农学-植物保护] 0901[农学-作物学] 090401[农学-植物病理学] 090402[农学-农业昆虫与害虫防治] 

基　　金：supported by the Zhejiang Provincial Natural Science Foundation of China (Grant No.Z305650 and No.Y3080361) the Science and Technology Department of Zhejiang Province,China (Grant No.2004C12020) the National Key Technology Research & Development Program,China (Grant No.2006BAD01A01-5 and No.2008ZX08001-001) the 151 Foundation for the Talents of Zhejiang Province,China 

主　　题：rice (Oryza sativa) Agrobacterium mixture-mediated co-transformation CrylAb gene Hpt gene gene transfer marker-free transgenic plants pest resistance 

摘      要：CrylAb gene was transformed into four rice varieties, Zhejing 22, Zhejing 27, Jiahua 1 and Xiushui 63 mediated by Agrobacterium-mixture co-transformation. Rice genotype had an important effect on callus induction and transformation efficiency. Different mixtures of Agrobacterium strains （EHA105 and EHA101） contained Hpt and CrylAb genes resulted in different frequencies of resistant calli. There was no correlation between the frequency of transformants with the ratio of the Agrobacterium strain mixture contained Hpt and CrylAb genes. A total of 509 transgenic plants were obtained from the four rice varieties, and 272 T2 progenies were analyzed for CrylAb and Hpt genes. PCR analysis revealed that 412 regenerated plants were Hpt positive （80.94%）, 62 plants were also CrylAb co-transformants （15.05% in total frequency）, and 42 plants among the 272 T2 progenies were CrylAb positive but Hpt negative. This suggests that marker-free transgenic plants could be produced by co-transformation mediated by mixed Agrobacterium strains with the selectable marker gene and target gene Southern blot analysis of five independent marker-free T2 transgenic lines co-transformed from Zhejing 22 showed that CrylAb gene had been inserted into rice genome with a single copy. The transgenic plants showed significantly stronger resistance to lepidopteron than the non-transgenic plants under no application of insecticides against lepidopteron.