Identification Of Nematode Species Of Genus Bursaphelenchus From Packing Wood Materials

Pine wilt disease has caused great damage to the forestry and ecological system in China. It became the most destructive threat to pine wood production in our country.Plant quarantine is one of important methods to prevent the disease. Some other Species of genus Bursaphelenchus are discovered from imported coniferous wrapper besides Bursaphelenchus xylophilus. Some of them are very similar to B.xylophilus by morphological characters. The following work was carried out by using eight samples of Bursaphelenchus spp. Intercepted from Port as researched material.Seven species of Bursaphelenchus spp. from coniferous wrapper were identified. They were B.xylophilus, B.mucronatus, B.rainulfi, B.conicaudatus, B.doui, B.singaporiensis, and B.ar..., respectively. B.rainulfi and B.conicaudatus were reported in 2000 and 2002, respectively. Recently, B.doui, B.singaporiensis and B. ar...have been identified as new species which have not been formally published yet (Private communication with Dr. Helen Braasch ). The sample wx5 has not been completely identified. This is first report about the identification of the above five species in China, In this work , species similar to B.xylophilus (B.doui , B.singaporiensis and B.conicaudatus) were compared with B.xylophilus.ITS regions of B. xylophilus , B. mucronatus and wx5 were analyzed. Amplification of ITS-PCR with universal primers showed that ITS of B. xylophilus and B. mucronats were amplified a ITS product with 940 bp and ITS product of wx5 is 1150 bp. As a results, ITS-PCR can be used to identify wx5 from B. xylophilus and B. mucronatus.Four restriction edonucleases were used to digest the ITS-PCR products of B. xylophilus and wx5. Digestion results of Alu I , Msp I , Hinf I andHae III revealed that interspecific digestion fragments were different. Sequence differences in the ITS region were used to synthesize specific primers to distinguish B. xylophlus from other species. When analyzing B. xylophilus, B. mucronatus and wx5 with the specific primer, only a PCR product with 580 bp was amplified from B. xylophilus.