Molecular diagnostics of some trichodorid nematodes and associated Tobacco rattle virus

Several Paratrichodorus and Trichodorus (trichodorid) nematode species are natural vectors of Tobacco rattle virus (TRV) and cause economically important diseases, especially in potato and ornamental bulbous crops. Identification of trichodorid species based on morphological characters is laborious, time-consuming, and requires the services of highly trained personnel. Molecular diagnostics for trichodorid nematodes, using the ribosomal DNA repeat unit, were successfully developed to distinguish two Paratrichodorus and two Trichodorus species. The complete sequences of the 18S genes and the ITS-1 regions for these species were obtained and species-specific primers successfully designed for them. An RT-PCR assay was developed utilizing isolate-specific primers that amplify serologically distinguishable strains of TRV in individual trichodorid nematodes. The primers were based on the highly conserved RNA-1 segment of the bipartite genome and also on different parts of the RNA-2 segment of the virus genome.