Investigation And RT-PCR Detection Of Wine Grape Virus Disease In Qinhuangdao

At present,China has reported the 14 kinds of grape viruses.The occurrence of viruses seriously restrict the development of grape industry in China.Qinhuangdao is one of the important wine-making viticulture areas in China,and there are few reports on the incidence of grape virus in Qinhuangdao.Therefore,This thesis was carried out on the following three aspects: First,Natural incidence investigation with wine grape virus in Qinhuangdao;Second,grape virus diseases RT-PCR detection;Thirdly,partial gene sequence analysis of GLRaV3 and GPGV Qinhuangdao isolates.The types of grape virus diseases were identified,which laid a foundation for the prevention and control of grape virus diseases in Qinhuangdao.The results of the study are mainly the following points.1.Five vineyards in Qinhuangdao for Grapevine fan leaf virus(GFLV),Grapevine leafroll-associated virus(GLRaVs),Grape virus B(GVB)and Grapevine fleck virus(GFKV)Natural infection situation was investigated.The results showed that GFLV and GLRaVs were widely distributed in the grape growing areas of Qinhuangdao,and the incidence of GLRaVs was the highest.GVB is only found in langes winery.No symptoms of GFKV were found in the vineyards surveyed.There were differences in the infection of GLRaVs among different grape plantations and varieties.The incidence of GLRaVs in gengzhuang grape plantation is higher than that of the other 4plantations;the GLRaVs of ’Merlot’ and ’Shiraz’ have higher infection rates.2.RT-PCR was performed on suspected symptom samples of 7 virus diseases in Qinhuangdao producing area.Among the tested samples,GFKV,GPGV,GLRaV3 and GVB were detected,among which GFKV,GPGV and GLRaV3 were investigated in all grapes,detection rates were 49.9%、59%、and47.3% respectively.The detection rate of GLRaV3 was the highest in the ’Petit Verdot’ variety,which was 82.2%;the detection rate of GFKV was the highest in the ’Petit Verdot’ variety,which was 84.4%;GPGV had the highest detection rate in the ’Muscat blanc’ variety,93.3%;GVB has the highest detection rate of ’Chardonnay’,which is 6.6%.In addition,all tested varieties were detected to contain double complex infection.On the basis of single virus RT-PCR detection,a dual RT-PCR detection system of GLRaV3 and GFKV was established,and the detection conditions were optimized.3.Sequencing analysis of GLRaV3-HSP70 and GPGV-MP genes obtained frompositive samples.The results showed that the nucleotide sequence homology of HSP70 gene of GLRaV3-QHD isolates with that of reported others was96.3%~99.3%.The nucleotide sequence homology of the GPGV-QHD isolate with the MP gene of the isolates reported at home and abroad was 93.3%~98.4%.