Application of Multiplex RT-PCR for Detection of Cucurbit-Infecting Tobamovirus

Cucumber green mottle mosaic virus (cgmmv) and kyuri green mottle mosaic virus (kgmmv) are seed borneviruses, and they are also transmitted mechanically during agricultural practice and through water. hence, theseviruses have potential diseases widely distributed throughout the world. to detect different strains of cgmmvand kgmmv, several specific primers for each virus were designed for single and multiplex rt-pcr. theresults of single and multiplex rt-pcr showed that cgmmv was detected in zucchini isolated in bali-indonesia, while kgmmv was detected both in zucchini isolated in bali-indonesia and cucumis metuliferusisolated in thailand. furthermore, artificial co-infection of these two viruses was prepared and carried out usingtwo different ways of viral rnas extraction. based on the results, it could be reported that viral rnas for cdnaamplification by multiplex rt-pcr could be extracted from a mixture of infected leaves or separate extractionof each viruses infected leaves. in addition, results presented in this study demonstrated the application ofmultiplex rt-pcr to simultaneously detect cgmmv and kgmmv from cucurbit leaves using a mixture offour primers and its feasibility as a sensitive and rapid laboratory assay. since, no multiplex rt-pcr techniquehas been described for the detection of cgmmv and kgmmv, this technique can be a good option for sensitiveand reliable tool for detection of two major cucurbit infecting tobamoviruses.