Detection and Identification of Bursaphelenchus Species with DNA Fingerprinting and Polymerase Chain Reaction

Judith H. Harmey, Matthew A. Harmey

Abstract


We have evaluated the potential of DNA-based methods to identify and differentiate Bursaphelenchus spp. and isolates. The isolation of a DNA probe, designated X14, and development of a DNA fingerprinting method for the identification and differentiation of Bursaphelenchus species and strains is described. Polymerase chain reaction (PCR) amplification of DNA isolated from Bursaphelenchus species using two primers derived from the sequence of the cloned repetitive DNA fragment X14 resulted in multiple band profiles. A 4-kb fragment thus amplified from B. xylophilus DNA was not amplified from B. mucronatus or B. fraudulentus DNA. In addition to this fragment, several other fragments are amplified from the three species. The banding patterns obtained allowed species identification and may have value in determining taxonomic affinities. Key words: Bursaphelenchus, differentiation, DNA fingerprinting, identification, nematode, pinewood nematode species complex (PWNSC), polymerase chain reaction, systematics, taxonomy.

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