Molecular Biological Diagnosis of Meloidogyne Species Occurring in Korea

Oh, Hyung Keun; Bae, Chang Hwan; Kim, Man Il; Wan, Xin Long; Oh, Seung Han; Han, Yeon Soo; Lee, Hyang Burm; Kim, Ik Soo

doi:2009.25.3.247

Plant Pathol J > Volume 25(3); 2009 > Article

The Plant Pathology Journal 2009;25(3):247-255.
DOI: https://doi.org/10.5423/PPJ.2009.25.3.247 Published online September 30, 2009.

Molecular Biological Diagnosis of Meloidogyne Species Occurring in Korea

Hyung Keun Oh, Chang Hwan Bae, Man Il Kim, Xin Long Wan, Seung Han Oh, Yeon Soo Han, Hyang Burm Lee, Ik Soo Kim

Abstract

Root-knot nematode species, such as Meloidogyne hapla, M. incognita, M. arenaria, and M. javanica are the most economically notorious nematode pests, causing serious damage to a variety of crops throughout the world. In this study, DNA sequence analyses were performed on the D3 expansion segment of the 28S gene in the ribosomal DNA in an effort to characterize genetic variations in the three Meloidogyne species obtained from Korea and four species from the United States. Further, PCRRFLP (Polymerase Chain Reaction-Restriction Fragment Length Polymorphism), SCAR (Sequence Characterized Amplified Region) PCR and RAPD (Randomly Amplified Polymorphic DNA) were also utilized to develop methods for the accurate and rapid species identification of the root-knot nematode species. In the sequence analysis of the D3 expansion segment, only a few nucleotide sequence variations were detected among M. incognita, M. arenaria, and M, javanica, but not M. hapla. As a result of our haplotype analysis, haplotype 5 was shown to be common in M. arenaria, M. incognita, M. javanica, but not in the facultatively parthenogenetic species, M. hapla. PCR-RFLP analysis involving the amplification of the mitochondrial COII and large ribosomal RNA (lrRNA) regions yielded one distinct amplicon for M. hapla at 500 bp, thereby enabling us to distinguish M. hapla from M. incognita, M. arenaria, and M. javanica reproduced via obligate mitotic parthenogenesis. SCAR markers were used to successfully identify the four tested root-knot nematode species. Furthermore, newly attempted RAPD primers for some available root-knot nematodes also provided some species-specific amplification patterns that could also be used to distinguish among root-knot nematode species for quarantine purposes.

Key Words: PCR-RFLP, RAPD, Root-knot nematodes, SCAR PCR, 28S rDNA sequence