Identification And Evolution Analysis Of Pepper Virosis In Hunan

Hunan is the important origin of the pepper, with the frequent transport of pepper trade expansion and around the seed, pepper virus diseases is becoming more and more serious, the impact of the pepper yield and product quality. There is a big difference between the main types of each region, many kinds of pepper virus disease, due to the regional climate, environment and other. Therefore, in order to understand the viral pathogens, to establish a rapid, accurate and sensitive virus detection technology for pepper production has important practical significance.In this study, Hunan pepper virosis samples by siRNAs deeping sequencing analysis and RT-PCR testing to verify that the main types of virus in Hunan are Tobacco mosaic virus (TMV), Cucumber mosaic virus(CMV), Potato virus Y(PVY), Broad bean wilt virus 2(BBWV2), Pepper mild mottle virus(PMMoV), Chilli veinal mottle virus(ChiVMV), Pepper vein yellows virus(PeVYV), Chilli ringspot virus(ChiRSV) and Tobacco mild green mosaic virus(TMGMV), PeVYV, TMGMV and ChiRSV were first report in Hunan,and mixed infection of this virus.1.the complete genome sequence of ChiVMV-HN and PMMoV-HNl Hunan isolate was obtained by cloning. ChiVMV-HN (GenBank accession number:KR296797 complete genome sequence is 971 Ont, don’t contain poly A.5’non coding region (5’-UTR) and 3’non coding region (3’- UTR) with 163 and 283 bp, respectively.164～9427 nucleotide sequences constitute a large ORF(open reading frame). The consistency of the complete genome of ChiVMV-HN were compared other same species was found 78.89%-99.07%.Establishment of phylogenetic tree revealed that ChiVMV is divided into 3 large groups, and the other Hunan isolate P1 (AY859497) genetic relationship recently, there may is a certain geographical correlation.PMMoV-HN1 (GenBank accession number:KP345899 complete genome sequences were 6356nt,5’non coding region (5’-UTR) and 3’non coding region (3’-UTR) with 69 and 198 bp, respectively. Four open reading frame (ORF), coding protein, respectively,126kDa protein,183kDa protein 28kDa protein and 17.5 kDa protein.Consistency between PMMoV-HN1 and other same species 94.2%-99.7%. Establishment of phylogenetic tree revealed that PMMoV is divided into two branch group,PMMoV-HN 1 (NCBI accession number:KP345899) and Beijing separation. PMMoV-CN(NCBI accession number:AY859497) phylogenetic relationship among recently.2.The ChiVMV, PMMoV, BBWV2 and PeVYV coat protein gene conservative sequence, designed to be able to simultaneously amplify the four kinds of virus specific primers. Four pairs of specificity primer determined by the Primer-BLAST were amplified out of 513,236,718 and 383 specific fragments, establishing a multiplex-PCR system for detection of ChiVMV, PMMoV, BBWV2 and PeVYV. The multiple PCR reaction system and the reaction conditions were improved, and the four viruses were detected simultaneously by multiplex-PCR reaction. The applicability of multiplex-PCR system was tested by 196 pepper samples.