The development and molecular characterization of a rapid detection method for Rice root-knot nematode (Meloidogyne graminicola)

The root-knot nematode Meloidogyne graminicola is a major constraint in rice production in the world. Using rDNA-ITS sequences data alignments, the genetic variation among twenty-one populations of M. graminicola (sixteen from Myanmar and five from China) was investigated. The results showed that all the populations were clearly separated from other species and that there was a low level of genetic variation among the isolates. A set of species-specific primers was designed to develop a species-specific molecular tool for the precise identification of M. graminicola. The primer reliability, specificity and sensitivity tests showed that the primer set (Mg-F3 and Mg-R2) amplified the expected fragment size of 369 bp from the template DNA of target nematode populations but not from non-target organisms. A duplex PCR test allows for saving diagnostic time and costs by amplifying the species of interest from a nematode mixture. Therefore, this species-specific primer set may be a powerful tool to improve taxonomic identification by non-specialists and the design of successful management practices as well.