Reliable identification of Bursaphelenchus xylophilus by rDNA amplification
AbstractPCR direct identification and PCR-RFLP analysis from single nematodes were employed to discriminate Bursaphelenchus xylophilus from B. mucronatus. For PCR direct identification, two specific primer sets from ITS amplified a 220 bp and 330 bp fragment from DNA of B. xylophilus, respectively. But no amplification band was obtained from DNA of B. mucronatus. Restriction patterns of PCR-RFLP have revealed that HinD and MspI could be used to discriminate B. xylopbilus from B. mucronatus. Both methods employed here could be used to identify single specimens of B. xylophilus sensitively and accurately. It is suggested that PCR direct analysis is more convenient than other techniques. PCR-RFLP analysis could be used as an alternate method to confirm the result of PCR direct analysis when necessary.