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Robust RNAi-Based Resistance to Mixed Infection of Three Viruses in Soybean Plants Expressing Separate Short Hairpins from a Single Transgene

文献类型： 外文期刊

作者： Zhang, Xiuchun 1 ; Sato, Shirley 3 ; Ye, Xiaohong 5 ; Dorrance, Anne E. 1 ; Morris, T. Jack 5 ; Clemente, Thomas E. 3 ;

作者机构： 1.Ohio State Univ, Dept Plant Pathol, Wooster, OH 44691 USA

2.Chinese Acad Trop Agr Sci, Key Lab Trop Crop Biotechnol, Minist Agr, Inst Trop Biosci & Biotechnol, Haikou 571101, Peoples R China

3.Univ Nebraska, Ctr Plant Sci Innovat, Lincoln, NE 68588 USA

4.Univ Nebraska, Dept Agron & Hort, Lincoln, NE 68588 USA

5.Univ Nebraska, Nebraska Ctr Virol, Lincoln, NE 68583 USA

6.Univ Nebraska, Sch Biol Sci, Lincoln, NE 68583 USA

关键词： double-stranded RNA;transgenes;35S promoter

期刊名称：PHYTOPATHOLOGY （ 影响因子：4.025； 五年影响因子：4.394 ）

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收录情况： SCI

摘要： Transgenic plants expressing double-stranded RNA (dsRNA) of virus origin have been previously shown to confer resistance to virus infections through the highly conserved RNA-targeting process termed RNA silencing or RNA interference (RNAi). In this study we applied this strategy to soybean plants and achieved robust resistance to multiple viruses with a single dsRNA-expressing transgene. Unlike previous reports that relied on the expression of one long inverted repeat (IR) combining sequences of several viruses, our improved strategy utilized a transgene designed to express several shorter IRs. Each of these short IRs contains highly conserved sequences of one virus, forming dsRNA of less than ISO bp. These short dsRNA stems were interspersed with single-stranded sequences to prevent homologous recombination during the transgene assembly process. Three such short IRs with sequences of unrelated soybean-infecting viruses (Alfalfa mosaic virus, Bean pod mottle virus, and Soybean mosaic virus) were assembled into a single transgene under control of the 35S promoter and terminator of Cauliflower mosaic virus. Three independent transgenic lines were obtained and all of them exhibited strong systemic resistance to the simultaneous infection of the three viruses. These results demonstrate the effectiveness of this very straight forward strategy for engineering RNAi-based virus resistance in a major crop plant. More importantly, our strategy of construct assembly makes it easy to incorporate additional short IRs in the transgene, thus expanding the spectrum of virus resistance. Finally, this strategy

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