Identification of Bursaphelenchus xylophilus and B.mucronatus by PCR-RFLP Technique

A ploymerse chain reaction-restriction fragment Length polymorphism(PCR-RFLP) analysis was used for discrimination of isolates of Bursaphelenchus xylophilus and B.mucronatus.The amplication of isolates of B.xylophilus yielded one fragment of approximately 890bp.But that of B.mucronatus was about 930bp.Digestion of amplified products of each nematode isolate with five restriction endonucleases revealed the results as follows:(1)Dra I digestion of its products of B.xylophilus populations yielded two fragments,510 and 380bp.But Dra I couldn't digest ITS products of B.mucronatus populations.(2)Sal I couldn't digest the ITS products of all B.xylophilus populations.But it could digest that of B.mucronatus populations to two fragments,which were 720 and 220bp;(3)Digesting production of four B.xylophilus populations by MspI yielded two fragments,530 and 360bp,except GZ02 which couldn't be digested.But B.mucronatus populations yielded three fragments,340,290 and(180bp);(4) All populations of B.xylophilus and B.mucronatus could not be digested by Apa I;(5) Digestions of amplified products of B.xylophilus and B.mucronatus with Xho I yielded two fragments respectively,520 and 370,530 and 400bp.The restriction endonucleases,Dra I and Sal I,could be used for identification of B.xylophilus and B.mucronatus.The results of digestion of B.xylophilus and B.mucronatus were such sharp difference that it was very easy to identify and it was very convenient to be applied,Msp I and Xho I were not fit for identification of B.xylophilus and B.mucronatus;Apa I could not distinguish and identify B.xylophilus and B.mucronatus.