黄瓜绿斑驳花叶病毒的生物学与分子检测鉴定

Biological and Molecular Testing Techniques for Cucumber Green Mottle Mosaic Virus

根据已报道的黄瓜绿斑驳花叶病毒(CGMMV)外壳蛋白(CP)基因序列合成1对引物,对侵染黄瓜的CGMMV广西分离物CP基因片段进行扩增和序列分析,结果表明该分离物与CGMMV-GX-G(DQ647384)同源性最高,达100%。将病毒分离物摩擦接种于5种葫芦科植物、普通烟、心叶烟及苋色藜上,结果表明在葫芦科植物上均表现症状,在苋色藜上产生局部侵染,不侵染普通烟和心叶烟。接种实验发现不同植物及接种时间对寄主的发病有一定影响。上述植物接种后进行RT-PCR检测,结果表明,从接种病毒的葫芦科植物上和苋色藜上均扩增到该病毒的特异性条带,而普通烟、心叶烟和健康对照没有扩增出目的条带。生物学与分子检测鉴定结果表明CGMMV-GX属于CGMMV西瓜株系。

Based on the reported nucleotide sequences of the coat protein（CP） gene of Cucumber Green Mottle Mosaic Virus（CGMMV）,a pair of primers was designed and used for cloning and sequencing CP gene fragment from the genome of CGMMV isolate which infected cucumber in Guangxi province,China.Results showed that the highest homology could reach 100% at the nucleic acid level with CGMMV-GX-G（DQ647384）.Mechanical inoculation test with the isolate was carried out and reverse transcription polymerase chain reaction（RT-PCR） was applied to detect and identify.Typical systemic symptoms were developed on those hosts of cucumber,watermelon,muskmelon,pumpkin and bottle gourd after inoculated with CGMMV,and local symptoms was found on Chenopodium amaranitcolor.No symptom was found on Nicotiana tabacum var.K326 and N.glutinosar.The result of analysis of variance proved that the different plant and inoculation time on the infection.The results of RT-PCR showed specific bands in the samples of inoculated cucurbitaceous plants while no bands were detected in the sample of Nicotiana tabacum var.K326,N.glutinosar and the untreated control group.Results of biological and molecular detection and identification showed that the isolate CGMMV-GX belonged to CGMMV W strains.