Identification Of Pathogens Causing Strawberry Virus Disease In Henan Province And Exploration Of Virus CDNA Clone Infection System

Strawberry is a kind of horticultural plant with high economic value and widely spreaded in the world.Since the runners and branches are the main origin of asexual propogation of strawberry,strawberry decline is easy to occur which shows the symptom of leaf distortion and stunting leading to the loss of yield and quality of strawberry,hence influences the economic value of strawberry.Currently the distribution of strawberry virus diseases in Henan province is unknown,thereby,in this study collecting strawberry diseased leaves in five areas of Henan province,to ascertain the mian pathoheny of causing strawberry virus diseases in Henan province,and build up the multiple PCR detection system of five strawberry virus diseases,at the same time,exploring the cDNA clone infect system of two main strawberry virus which library have owned,in order to provide theoretical basis and technological support for controlling strawberry virus diseases of Henan province.The mian experimental result as following:Through the acquisition of Zhengzhou city,Xinxiang,Luoyang,Jiaozuo and Pingdingshan Strawberry leaf disease,Strawberry virus specific primers to make use of 11 RT-PCR test,the results showed that in Zhengzhou caused Strawberry virus disease pathogen mainly with Strawberry vein banding virus,Strawberry necrotic shock virus,Strawberry pallidosis-associated virus.The main pathogens of strawberry virus disease caused by xinxiang city are as follows Strawberry mottle virus,SVBV,SPaV。The main pathogens causing strawberry virus disease in pingdingshan are SVBV and SMYEV.The main pathogens causing strawberry virus disease in jiaozuo city are SVBV,which can be detected in the above five regions.Compound infection of SMoV and SVBV was found in luoyang city.Compound infection of SVBV and SMYEV was found in pingdingshan city.Based on SPaV,which is the first discovery in China,the nearly full-length sequence of SPaV was obtained by piecewise amplification,with RNA1 of 8066bp and RNA2 of 7978bp.Phylogenetic analysis was carried out by amplifying the cp full-length reading frame of different SVBV isolates,and the results showed that the 5 isolates and other isolates in China were all clustered into a large branch,the Canadian isolates were isolated,and the Zhengzhou isolates were most closely related to the Hefei.By amplifying the cp full-length reading frame of SNSV,systematic analysis was carried out.The results showed that the SNSV isolates in zhengzhou were closely in Florida,the United States,and clustered into a large branch with the isolates in the United States and Australia,while the Japanese isolates formed a branch separately.In addition,according to the obtained sequences of 5 strawberry viruses,specific primers were designed and the multiple PCR amplification system of 5 strawberry viruses was established through fumble annealing temperature and time.The amplification lengths were SVBV 816 bp,SMoV 620 bp,SMYEV 444 bp,SPaV 276 bp and SNSV 134 bp,respectively.Inoculate SMYEV and SMoV through vacuum filtration and friction to noculate SMYEV and SMoV,the results show that:friction inoculation efficiency of SMoV was 2/3 and 1/3,respectively,vacuum inoculation efficiency of SMoV was 1/4 and 1/2,respectively,the vacuum inoculation efficiency of SMYEV was 1/6 and 2/11,respectively.Nicotiana benthamiana which be infiltrated by agrobacterium,showed that both viruses could infect the inoculated leaves but could not be detected in the system leaves.