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Acta Agron Sin ›› 2013, Vol. 39 ›› Issue (09): 1594-1601.doi: 10.3724/SP.J.1006.2013.01594

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Agrobacterium tumefaciens Mediated Transformation of RNAi CP Gene into Soybean (Glycine max L.)

ZHANG Jie-Qiong1,LI Hong-Yan1,HU Xiao-Nan1,SHAN Zhi-Hui2,*,TANG Gui-Xiang1,*   

  1. 1 Institute of Crop Science, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou 310058, China; 2 Oil Crops Research Institute, Chinese Academy of Agriculture Sciences, Wuhan 430062, China
  • Received:2012-11-28 Revised:2013-05-24 Online:2013-09-12 Published:2013-07-09
  • Contact: 唐桂香, E-mail: tanggx@zju.edu.cn; 单志慧, E-mail: zhihuijimi@163.com

Abstract:

 Soybean mosaic virus (SMV) causes a severe disease in soybean, which can be efficiently prevented by planting resistant cultivars. In order to improve the SMV resistance of soybean, an agrobacterium-tumefaciens mediated gene transformation was conducted in this study by using RNAi soybean mosaic virus coat protein (CP)gene as the expression vector, bar gene as the selective marker gene, and cotyledonary-node as the explant to get RNAi transformation soybean. As results, 22 putative transgenic soybean plants were obtained and 18 positive transgenic soybean plants were identified by coating with leaves herbicide, using bar protein quick dip stick and PCR analysis. The segregation ratio of T1 transgenic progeny showed that the transformed gene could be inherited according to the Mendel's law. The T1 southern blot analysis showed that the imported interference fragment was one copy. After SMV friction inoculation, the RNAi CP transgenic soybean plants showed good resistance to SMV. DAS-ELISA analysisat three weeks after SMV inoculation revealed that 100% of the non-transgenic lines SMV while only 7.69% of the RNAi plants were infected by SMV. The results demonstrated that the RNAi CP transgenic soybean plants obtained in the study are valuable resources for improving the SMV resistance of soybean.

Key words: Soybean, RNAi CP, Genetic transformation, Soybean mosaic virus, Virus identification

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