The Study On Effect Of OTU Gene Of VdCVl On The Plant Virus As Well As Transgene Verticillium Dahliae

Verticillium dahliae chrysovirus1(VdCV1) was found from the Verticillium dahlia in2011,and deduced that it contained an Ovarian Tumor protease gene, however the function of the OTUgene has not confirmed yet. So, in this paper, it was studied that effect of OTU gene of VdCV1on the plant virus as well as transgenic Verticillium dahliae in order to ascertain its effect on theinteraction functions among VdCV1-Verticillium dahlia-Cotton.First, on study on the OTU gene’s effect on plant virus, we use viral vector (FoMV) andcucumber mosaic virus(CMV) as basic frame to expressing foreign gene OTU, and get the viralexpression vector pCB301-FoMV-OTUp193and pCB301-F209-Δ2b-OTUp193. Then in orderto study the effects of OTU gene expression to FoMV RdRp replicase and the GFP expression ofthe recombinant virus CMVand pCB301-FoMV-Δ2,3,4,5, pCB301-FoMV-GFP,pCB301-FoMV-OTUp193, pCB301-F209-Δ2b, pCB301-F209-Δ2b-OTUp193andpCB301-F209-Δ2b-GFP were transformed into Nicotiana benthamiana, respectively, by someagrobacterium infiltration experiments. It shows that, the OTU gene have an stabilization effectto RNA of RdRp of FoMV, and it improved the expression of GFP of recombinant FoMV-GFPand CMV(F209-Δ2b-GFP,F109,F309). In short, the OTU gene demonstrated an enhancementeffect on viral replication. Furthermore, we have analysis the expression of pathogenesis-relatedgenes NtPR1a in different experimental treatment. The results showed that the OTU gene has aninhibition effect on the NtPR1a gene, and it indicated the OTU gene decreased the immunity ofplants.On the study on transgenic Verticillium dahlia, we put OTU gene and small fragments ofOTU gene—OTUp193gene inserted into the vector pHMG. And we named them thepHMG-OTU and pHMG-OTUp193vectors. In addition, the OTU and GFP gene also insertedinto the vector pBHt1. Then using the methods of Agrobacterium tumefaciens-mediatedtransformation, pHMG, pHMG-OTU, pHMG-OTUp193, pBHt1and pBGFP were transformedinto the Verticillium dahliae T-9. The results o show that we have gain the transformant ofVerticillium dahliae with GFP gene, and proved that the Pmg1promoter can be used in thegenetic transformation of Verticillium dahlia. The expression vectors with OTUgene(pHMG-OTU and pHMG-OTUp193) and pBHt1were also successfully transformed into Verticillium dahliae and the transformants are detected by genome DNA PCR. However, it isstill need further research for this result.