Journal of Taiwan Agricultural Research

Author:Hui-Fang Ni, Ruey-Shyang Chen, Mei-Hwa Wang, Tung-Tsuan Tsay and Yung-Hsiung Cheng*

Abstract:

    The main objective of the study is to design a pair of specific primers for the detection of Meloidogyne spp. by PCR. The internal transcribed spacer (ITS) region of the ribosomal DNA from the representative isolates of M. incognita, M. javanica, M. arenaria, and M. graminicola were amplified by universal primers. The entire ITS region, including 5.8S subunit, was cloned and sequenced. The aligned results of obtained sequences showed that a high level of sequence identity was found among M. incognita, M. javanica, and M. arenaria. Two PCR primers, Mi148 and Mi452, were designed based on rDNA sequence of M. incognita. The primer pair was subsequently shown to amplify a 342 bp fragment from the DNA of Meloidogyne spp. On the contrary, this primer pair failed to amplify any fragments from the DNA from other tested nematodes. Using this primer pair, Meloidogyne spp. could be successfully detected from diseased plants and soil. The detection limit of the primer pair was as few as 10 pg template DNA, or 5 juveniles in soil.

Key words:Meloidogyne spp., Plant pathogenic nematodes, PCR-mediated detection, Ribosomal DNA (rDNA), Internal transcribed spacer (ITS)

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