Abstract:

The pine wood nematode (Bursaphelenchus xylophilus) is a highly damaging pest on a world scale. Monochamus alternatus is the most important vector of B. xylophilus. A quick, reliable and replicable method is needed for detecting the pest. To that end, we firstly developed a simple procedure for isolating DNA from the mixture of nematode +beetle tissue for subsequent nematode detection by PCR amplification. Then one pair of B. xylophilus-specific primers that generated a specific amplicon of 403 bp (DQ855275) located respectively in the ITS1 and ITS2 regions， were choosen. No amplicon was obtained from either M. alternatus or any of the 12 other nematodes species related to pine wilt disease (B.mucronatus, B.aberrans, B.corneolus, B.leoni, B.hunanensis, B.teratospicularis, Aphelenchoides resinosi, Seinura steineri, Ditylenchus parvus，Odorhabdiplogaster xiphocaudatus, Rhabditida sp. And Parasitorhabditis sp.) with the primers. The same 403 bp amplicon was also firstly obtained by PCR amplification from the beetle tissue infested with B. xylophilus.

Key words: Bursaphelenchus xylophilus, beetle tissue of Monochamus alternatus, specific PCR primers