مقایسه مولکولی جمعیت‌هایی از Meloidogyne javanica و Meloidogyne incognita در ایران با روش PCR – RFLP

To carry out this study, total DNA was extracted from eggs and from second stage juveniles of several populations of Meloidogyne javanica and Meloidogyne incognita, using phenol / chloroform method. Following extraction, DNA was electrophoresed on 1% agarose gel to determine its quality and quantity. A specific primer pair (C2F3 / 1108 23 and 20 nucleotides, respectively) was used to discriminate M. javanica from M. incognita populations using polymerase chain reaction (PCR). Primer annealing sites were located in the 3′ portion of mitochondrial gene encoding cytochrome oxidase subunit II (COII) and in the 16S rRNA gene. Following PCR amplification, electrophoresis of amplified DNA showed 1.7 kb fragment in populations of both species. Digestion of 1.7 kb amplified product with HinfI restriction endonuclease resulted in the generation of two DNA fragments of 0.7 and 1.0 kb in M. javanica and three DNA fragments of 0.3, 0.4 and 1.0 kb in M. incognita. There were no differences in the digestion patterns among various populations of each species examined.