RNAi-mediated SMV-P3 Silencing Increases Soybean Resistance to Soybean Mosaic Virus

YANG Xiang-Dong1 NIU Lu1 ZHANG Wei1 YANG Jing1 DU Qian1 XING Guo-Jie1 GUO Dong-Quan1 LI Qi-Yun1 DONG Ying-Shan1

(1.Agricultural Biotechnology Institute, Jilin Academy of Agricultural Sciences/Jilin Provincial Key Laboratory of Agricultural Biotechnology, Changchun, China 130033)

【Abstract】 Soybean mosaic virus (SMV) is one of the most important diseases in major soybean production areas, which has severe effects on soybean production and seed quality in China. Cultivation of disease-resistant varieties is the most economical and effective strategy to prevent and control SMV. In this study, RNAi fragments of the gene encoding P3 protein, which is involved in SMV mobility and affecting host range, were introduced into soybean by plant-mediated RNA interference (RNAi) techniques to explore the influence of RNAi-mediated SMV-P3 silencing on soybean SMV resistance. Southern blot analysis revealed that exogenous RNAi fragments were integrated into the soybean genome at low copy numbers (1–4). T1–T3 generation transgenic soybeans were sprayed with herbicide and inserted fragments were examined using PCR. The results indicated that T-DNA insertion fragments could be stably inherited between generations of transgenic soybean. Inoculation of T2and T3 generation transgenic soybeans with SMV suggested that transgenic soybeans exhibited significantly higher resistance to the prevailing SMV strain, SC-3 in major soybean production areas than the non-transgenic control varieties Williams 82 and SN9. The disease index was reduced by 4.37–18.51. Furthermore, the resistance could be stably inherited. In conclusion, RNAi-mediated SMV-P3 silencing can significantly increase the SMV resistance of the transgenic soybean. Soybean mosaic virus(SMV)is one of the most important diseases in major soybean production areas and has severe effects on soybean production and seed quality in China.Breeding disease-resistant varieties is the most economical and effective strategy to prevent and control SMV.In this study,RNAi fragments of the gene encoding P3 protein,which is involved in SMV mobility and affecting host range,were introduced into soybean by plant-mediated RNA interference(RNAi)techniques to explore the influence of RNAi-mediated SMV-P3 silencing on soybean SMV resistance.Southern blot analysis revealed that exogenous RNAi fragments were integrated into the soybean genome at low copy numbers(1–4).T1–T3generation transgenic soybeans were sprayed with herbicide and inserted fragments were examined using PCR.The results indicated that T-DNA insertion fragments could be stably inherited between generations of transgenic soybean.Inoculation of T2and T3generation transgenic soybeans with SMV suggested that transgenic soybeans exhibited significantly higher resistance to the prevailing SMV strain,SC-3in major soybean production areas than the non-transgenic control varieties Williams 82 and SN9.The disease index was reduced by 4.37–18.51.Further,the resistance could be stably inherited.In conclusion,RNAi-mediated SMV-P3 silencing can significantly increase the SMV resistance of the transgenic soybean.

【Keywords】 Soybean; Soybean mosaic virus; SMV-P3; RNAi-mediated gene silencing;

【DOI】

【Funds】 China National Major Project for Developing New GM Crops (2016ZX08004-004) supported by the China National Major Project for Developing New GM Crops(2016ZX08004-004) Science & Technology Development Project of Jilin Province (20150204011NY) the Science&Technology Development Project of Jilin Province(20150204011NY)

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This Article

ISSN:0496-3490

CN:11-1809/S

Vol 42, No. 11, Pages 1647-1655

November 2016

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Abstract

  • 1 Materials and methods
  • 2 Results and analysis
  • 3 Discussion
  • 4 Conclusions
  • References