Biological and Molecular Testing Techniques for Cucumber Green Mottle Mosaic Virus

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.