Identification of Japanese species of the genus Meloidogyne (Nematoda: Meloidogynidae) by PCR-RFLP analysis

The polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis for identification of ten Meloidogyne species from Japan was attempted on an individual second-stage juvenile. The PCR was performed with the primer set to amplify the region between the cytochrome oxidase subunit II gene and the large subunit of the 16S ribosomal RNA gene in mitochondrial DNA. A single fragment of about 1.7 kb was produced in M. incognita, M. javanica and M. arenaria. The sizes of amplified products differed in M. hapla, M. suginamiensis, M. marylandi, M. mali, M. camelliae, M. sp. near mall and M. sp. near arenaria. The fragment size of each product ranged from 0.5 kb to 0.6 kb. DraI, MseI, SspI and VspI produced digested fragments in all ten Meloidogyne species. Though a SspI or VspI + HinfI-digested pattern separated the ten Meloidogyne species into nine groups, we were unable to distinguish M. arenaria from M. javanica using these patterns. MseI-digested pattern alone distinguished M. arenaria from M. javanica. MseI and either SspI or VspI + HinfI-digested patterns of the PCR product are most suitable for discrimination of these ten Meloidogyne species from Japan.