利用病毒诱导的基因沉默技术沉默TSWV的NSm基因

Using Virus-induced Gene Silencing to Silence the NSm Gene of TSWV

【目的】番茄斑萎病毒(Tomato spotted wilt orthotospovirus,TSWV)在全球范围内严重危害农业生产,病毒诱导的基因沉默(virus-induced gene silencing,VIGS)技术主要用于植物内源基因的功能研究,而其沉默植物外源病毒基因用于病毒防控鲜有报道。本研究利用VIGS技术在辣椒中沉默植物外源病毒TSWV的NSm基因,为辣椒产业防控TSWV病害奠定技术基础。【方法】以辣椒(Capsicum annuum L.)为研究材料,首先构建辣椒PDS基因的沉默载体pTRV2-PDS以评估该沉默载体在辣椒中的有效性,再构建TSWV NSm基因的沉默载体pTRV2-NSm评估该沉默载体对TSWV NSm基因的沉默效果。【结果】构建的沉默载体pTRV2-PDS在辣椒中有效,沉默载体pTRV2-NSm可以高效沉默TSWV的NSm基因,NSm的表达量仅为对照组的4.6%,且表现出对TSWV的抗性。【结论】构建的沉默载体pTRV2-NSm可以沉默TSWV的NSm基因,并使辣椒对TSWV产生一定的抗性。

[Purpose]Tomato spotted wilt orthotospovirus(TSWV)has seriously endangered agricultural production around the world.Virus-induced gene silencing(VIGS)is mainly used for the functional research of endogenous genes on plant and the use of VIGS to silence exogenous virus genes for plant protection against virus is rarely reported.In this study,VIGS was used to silence the NSm gene of TSWV on pepper in order to lay a technical foundation for the prevention and control of TSWV disease in pepper planting.[Method]Peper(Capsicum annuum L.)was selected as the re-search material.The silencing vector pTRV2-PDS for pepper PDS gene was constructed to evaluate the effect of the silencing vector in pepper.The silencing vector pTRV2-NSm was constructed to si-lence NSm gene of TSWV and protect pepper from TSWV.[Result]The pTRV2-PDS vector could effectively silence the PDS gene in pepper.The pTRV2-NSm vector can efficiently silence the NSm gene of TSWV on pepper.Inoculation of pTRV2-NSm made the expression of NSm in pepper only 4.6%of that in the control group,and provided resistance to TSWV.[Conclusion]This study showed that pTRV2-NSm vector could silence the NSm gene of TSWV on peppers,and give pepper resistance against TSWV.