番茄斑萎病毒N基因的VIGS载体构建

Construction of a VIGS vector containing N gene derived from tomato spotted wilt orthotospovirus

番茄斑萎病毒(tomato spotted wilt orthotospovirus,TSWV)严重危害多种经济作物和园艺植物,其N基因与病毒的侵染力密切相关,将N基因直接构建到VIGS载体上研究其致病功能目前鲜见报道。本研究将TSWV N基因构建到pTRV-PTV00载体上,注射本氏烟,通过qRT-PCR定量分析发现先接种TSWV后注射沉默载体的植株抗TSWV效率达57.60%,先注射沉默载体后接种TSWV的植株抗TSWV效率达99.14%。结果表明先注射载体后接种TSWV的N基因沉默效率更高。TSWV N基因VIGS载体的构建可为研究N基因在病毒致病等方面的功能提供前期材料,并为TSWV抗病育种和田间绿色防控提供一定的理论依据。

Tomato spotted wilt orthotospovirus(TSWV)causes serious diseases of many economically important plants such as cash crops and horticultural flora,TSWV contains a N gene that has strong effects on its infection,but there is no report about construction of a VIGS vector containing N gene to study its pathogenic function.In this study,the N gene of TSWV was inserted into pTRV-PTV00 vector and their combinant vector was then injected into N.benthamiana to study VIGS efficiency.Quantitative real-time PCR(qRT-PCR)analysis showed that the anti-TSWV efficiency of the plants inoculated with TSWV before injected the constructed vector was 57.60%,while the plants injecting constructed vector before inoculated with TSWV showed anti-TSWV efficiency of up to 99.14%.The results indicated that injecting constructed vector before inoculated with TSWV enhance silencing efficiency significantly.The constructed vector can be used for studying the function of N gene,for example,in pathogenesis,and the obtained information providing theoretical basis for TSWV control in fields.