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IDENTIFICATION OF PRATYLENCHUS COFFEAE AND P. LOOSI USING SPECIFIC PRIMERS FOR PCR AMPLIFICATION OF RIBOSOMAL DNA BY T. UEHARA1), T. MIZUKUBO2), A. KUSHIDA1) and Y. MOMOTA1) 1)Laboratory of Plant Nematology, Hokkaido National Agricultural Experiment Station, Hitujigaoka 1, Toyohira-ku, Sapporo 062-8555, Japan; 2)Laboratory of Plant Nematology, Kyushu National Agricultural Experiment Station, Nishigoshi, Kikuchi-gun, Kumamoto 861-1192, Japan The 18-26S region of rDNA of P. coffeae and P. loosi were amplified by PCR from genomic DNA. Sequence analyses indicated that the 5.8S gene sequences were conserved whereas the internal transcribed spacer (ITS) sequences were divergent. Sequence differences in the ITS were used to synthesize specific primer sets to differentiate P. coffeae and P. loosi. PCR assays with these primers specifically amplified a characteristic DNA fragment from each species. Moreover, the same specific amplification products were obtained using DNA extracted from single females males or juveniles. Keywords: identification, ITS sequences, PCR, rDNA. Root-lesion nematodes, Pratylenchus spp., are cosmopolitan and economicall) important migratory endoparasitic nematodes (Jatala & Berge, 1990). More than 20 Pratylenchus species are known in Japan (Mizukubo, 1992). Some of these, e.g. P. penetrans (Cobb) Filipjev & Schuurmans Stekhoven, P. coffeae Zimmer- man, P. vulnus Allen & Jensen and P. loosi
Nematologica – Brill
Published: Jan 1, 1998
Keywords: PCR; identification; ITS sequences; rDNA
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