Investigation And Identification Of Vegetable Virus Diseases In Liaoning Province

Vegetable virus diseases have become one of the most impotant factors that restrict the development of vegetable industry. It can cause a great impact on the quality and output. However, up to now, our understanding of the species, distribution, variation of the vegetable viruses were limited in Liaoning province. Some important virus such as Tomato yellow leaf curl virus, Pepper mild mottle virus, Cucumber mosaic virus and viruses on okra and Cryptotaenia japonica should be pay attention to. The survey and identification of vegetable virus were studied systematically in this article and reference was provided for future prevention and treatment of vegetable virus disease. Results showed as follows:1. The samples of vegetable viruses disease were collected and tested. The distribution characteristics of vegetable virus diseases were clarified in Liaoning province.2015, the 634 samples were collected from Solanaceae (tomato, pepper), Cucurbitaceae (cucumber), Leguminosae (bean), Malvaceae (okra) and Umbelliferae (Cryptotaenia japonica.), including the common vegetables, and health vegetables and wild vegetables in 22 towns, Tieling, Jinzhou, Xinmin, Haicheng, Fengcheng in Liaoning Province. The monoclonal antibody of Tobacco mosaic virus, Cucumber mosaic virus, Broad bean wilt virus, Tomato spotted wilt virus, Cucumber green mottle mosaic virus were used in Dot-ELISA method. TMV, CMV, BBWV, TSWV, PVY and other types of viruses were detected in Solanaceae vegetables. BBWV and TSWV were detected in Leguminosae vegetables. CMV and CGMMV were detected in Cucurbitaceae vegetables. The results showed that 117 samples were infected by viruses, the detection rate was 18%. Among them, TMV, CMV, PVY, BBWV and TSWV were detected and the average detection rate was 11.2%,9%,2.8%,2.4% and 0.9% separately. But CGMMV not be find in Cucurbitaceae vegetables. From the view of various crops, Solanaceae vegetables have higher virus rate. The detection rate was 23.5%, Cucurbitaceae and Leguminosae vegetables viruses detection rate were 12.8% and 13% respectively. There was many kinds of vegetable viruses in Fengcheng city. In the survey,12 samples were detected to have compound infection, the detection rate was 1.9%. Compound infection of TMV+CMV+BBWV+TSWV, TMV+BBWV and TMV+CMV.In the survey, TYLCV and PMMoV were investigation because that these viruses have continued to spread, the expansion of the trend of in Liaoning province in recent years. At the same time viruses were identified on okra and Cryptotaenia japonica in order to provide reference for the research and prevention of this kind of virus disease in the future.2. The molecular identification of TYLCV was carried on in order to clear the distribution in Liaoning Province. Samples were collected in many cities, such as Haicheng, Jinzhou, Shenyang, Tieling, Fengcheng and Fuxin. Whole genome of isolates were amplified by PCR used back to back primers. The results showed the samples in Haicheng were infected by TYLCV. The nucleotide sequence homology of TYLCV Haicheng isolate and TYLCV-SDCL (JQ038240.1) Shandong isolate was 99.35% without variation. In the phylogenetic tree and other domestic and foreign isolates belong to the same branch may have the same evolutionary ancestors.3. PMMoV was identification by biological, serological and molecular biological method. There are local spots in N. tabacum var. samsun, Nicotiana tabacum, N. glutinosa and chlorisis spots in Chenopodium amaranticolor. Maybe PMMoV infection in pepper by PMMoV preliminary judgment. By DAS-ELISA and RT-PCR detection, the results were PMMoV. Cloned PMMoV CP gene sequence, found that PMMoV isolates and PMMoV-BD(HQ699079) highest homology was 99.79%, and the phylogenetic evolution belong to a tree.4. The identification of viruses on okra and Cryptotaenia japonica were carried on by biological, serological and molecular biological methods. Okra and Cryptotaenia japonica became important health vegetable and have broadly market prospect for the rich nutritional value. Biological results showed symptom on Nicotiana tabacum, Gomphrena globosa and Chenopodium amaranticolor. According to symptoms judgment okra and Cryptotaenia japonica were infected by TMV. DAS-ELISA and sequence analysis showed that both of them were infected by TMV. Okra TMV isolates and Kunming Yunnan (HE818410.1) highest homology was 100%, and the phylogenetic evolution belong to a tree; Cryptotaenia japonica. TMV isolates and Kunming Yunnan (HE818410.1) and Guizhou (HE818450.1) highest homology was 100% and the phylogenetic evolution belong to a tree.