Molecular diagnosis and differentiation of Meloidogyne arenaria, Meloidogyne javanica and Meloidogyne incognita using SNP-based KASP assays

Root-knot nematodes (RKNs) are one of the most economically important plant-parasitic nematodes in many crops. Meloidogyne arenaria, M. incognita and M. javanica are the most common species attacking tomato, pepper, eggplant, cucumber, melon, watermelon and zucchini, which have economic importance in vegetable growing areas. Thus, a rapid and accurate identification system is required for RKNs. In this study, Kompetitive Allele-Specific PCR (KASP) genotyping assays were developed to identify three common RKNs. The novel technique developed in this study utilized only two primers: one allele-specific forward primer and a reverse common primer. Meloidogyne arenaria, M. incognita and M. javanica were separated easily from each other and other RKN species. This is the first report on three species identified by the KASP assay. The results showed that the KASP assays are suitable for the high-throughput identification of these species. The developed system can be used to identify three RKN species in research, routine and quarantine laboratories.