To overcome limitations in the morphological identification of Bursaphelenchus species, a molecular approach was used. The internal transcribed spacer 1 (ITS1) of the nuclear ribosomal DNA (rDNA) region was amplified and sequenced. A pairwise comparison of this sequence data revealed that the differences in the ITS1 region (approximately 308 bp) between four B. xylophilus isolates were slight (only 1 bp), but there are marked differences (up to 7 bp) in this region between the B. mucronatus isolates from China (Guangdong) and France.The data also revealed that the differences in the ITS1 region between the two Bursaphelenchus species range from 32-39 bp. Based on these data, the method of extracting the genomic DNA of the individual pinewood nematode and PCR-SSCP (single-strand conformation polymorphism) analysis were employed for the unequivocal differentiation of the Bursaphelenchus variant from B. xylophilus and B. mucronatus. These methods should provide a valuable tool for sensitive and accurate identification of individual worm of B. xylophilus.
NETL
NSTL
维普
万方数据
