VIGS结合HIGS诱导的烟草对斜纹夜蛾与棉铃虫的抗性比较

Comparative Study on the Resistance of Tobacco Plants Induced by VIGS and HIGS to Spodoptera litura and Helicoverpa armigera

为探索防控斜纹夜蛾和棉铃虫的新途径,比较研究了病毒诱导表达昆虫NDUFV2(NV2)基因片段的烟草植株对斜纹夜蛾与棉铃虫的抗性。以含SliNV2370或HaNV2370基因片段的CMV或TRV重组质粒-农杆菌,采用质粒-农杆菌浸润法接种本氏烟植株,将RT-PCR检测接种成功的本氏烟植株用于斜纹夜蛾或棉铃虫抗性试验,并采用RT-qPCR分析斜纹夜蛾和棉铃虫NV2基因表达量。结果表明:通过CMV载体或TRV载体表达SliNV2370基因片段的本氏烟植株都未能显著增强对斜纹夜蛾的抗性;斜纹夜蛾SliNV2基因表达量下降了19%左右,但沉默SliNV2基因的效果都没有达到差异表达基因的水平。CMV载体或TRV载体表达HaNV2370基因片段的本氏烟植株,都能显著增强对棉铃虫的抗性,分别提高36.5%和31.5%。棉铃虫HaNV2基因表达量分别下降了72.5%和78.7%,沉默HaNV2基因的效果都达到了差异表达基因的水平。说明通过CMV载体或TRV载体表达NDUFV2(NV2)基因片段的本氏烟植株未能显著增强对斜纹夜蛾的抗性,但能显著增强对棉铃虫的抗性。

Spodoptera litura and Helicoverpa armigera are important agricultural pests and widely distributed in the world.To develop new methods to control Spodoptera litura and Helicoverpa armigera,a study on the resistance of tobacco plants expressing NDUFV2(NV2)fragment induced by the virus vector was carried out.Agrobacterium-mediated infectious cDNA clones of CMV and TRV containing gene fragment SliNV2370 or HaNV2370 were constructed respectively,and the Nicotiana benthamiana plants were inoculated by agrobacterium infiltration method.The expressions of the virus and the target gene fragments in N.benthamiana were detected by RT-PCR.The successfully inoculated plants were used to study the resistance to Spodoptera litura or Helicoverpa armigera,and the NV2 gene expressions of Spodoptera litura and Helicoverpa armigera were analyzed by RT-qPCR.The results showed that the expression of SliNV2370 by CMV or TRV vector did not significantly enhance N.benthamiana plants resistance to Spodoptera litura,though the silencing effect of SliNV2 gene was about 19%which did not reach the differential expression level.The expression of HaNV2370 by CMV or TRV vector significantly enhanced N.benthamiana plants resistance to Helicoverpa armigera by 36.5%and 31.5%,respectively,and the expressions of HaNV2 gene were significantly reduced by 72.5%and 78.7%,respectively.Taking together,we concluded that the expression of NDUFV2(NV2)gene fragment by CMV or TRV vector did not significantly enhance N.benthamiana plants resistance to Spodoptera litura,but it significantly enhanced N.benthamiana plants resistance to Helicoverpa armigera.