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《Scientia Agricultura Sinica》 2013-06
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Silencing Rice Stripe Virus Gene in Laodelphax striatellus(Fallén) by dsRNA Feeding

CHEN Ai-ling1,2,LIN Ke-jian1,HE Kang1,2,WANG Gui-rong1,LI Fei2(1State Key Laboratory for Biology of Plant Diseases and Insect Pests/Institute of Plant Protection,Chinese Academy of Agricultural Sciences,Beijing 100193;2College of Plant Protection,Nanjing Agricultural University/Key Laboratory of Crop Pest Integrated Management,Ministry of Education,Nanjing 210095)  
【Objective】 The objective of this study is to use RNA interference(RNAi) technique to explore the feasibility of inhibiting the reproduction of Rice stripe virus(RSV) and to probe the interactions among different RSV genes in vitro.【Method】 The inhibition function of exogenous dsRNAs to the expression of RSV genes in Laodelphax striatellus(Fallén) were investigated through feeding synthesized dsRNA in vitro.According to the sequences of RSV,three types of dsRNAs(dsRdRp,dsNSvc4,dsCP) were designed and synthesized and then used to feed 2nd instar L.striatellus that infested with RSV.The dsRNA were synthesized by using the T7 RiboMax Express RNAi System.Then 1 μg?μL-1 dsRNA was added to the artificial diet for feeding.The expression level of each RSV gene in insects was examined by quantitative real-time PCR(qRT-PCR) at 96th hour after dsRNA treatment.【Result】 The results of qRT-PCR indicated that the expression of two RSV genes,NSvc4 and CP,decreased by 93.2% and 94.9%,respectively.Although the decreasing amplitude of RdRp gene was significantly lower,it still decreased by up to 45.6%.Further studies showed that the expressions of other non-target RSV genes decreased when feeding specific dsRNA,which indicated that interaction might exist among different RSV genes.【Conclusion】 This study offered an important basis for controlling RSV through dsRNA microbe engineering or transgenic rice technology.
【Fund】: 国家“973”计划项目(2010CB126200);; 国家自然科学基金项目(31272042)
【CateGory Index】: S435.111.4
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