松材线虫体外酶组成分析

The Enzymes in the Secretions of Pine Wood Nematode ( Bursaphelenchus xylophilus )

将表面消毒的松材线虫，在Ｔｒｉｓ－ＨＣｌ缓冲液中振荡培养，培养液经硫酸铵沉淀、透析、冷冻真空干燥，制成的干样，用于聚丙烯酰胺凝胶电泳。结果表明，松材线虫能向体外分泌多种可溶性蛋白，对其中酶组成分析发现有纤维素酶、过氧化物酶、蛋白酶、淀粉酶等。同样条件下未能检出β－半乳糖酶、β－木糖酶、β－葡聚糖酶。

After surface sterilization in the antibiotic solution of 0.002% actidion and 0.1 % sulphostreptomycete,PWN, Bursaphelenchus xylophilus, were in the buffer of Tris HCl,0.05M,pH7.2,for shaking incubation at 25 ℃ for 72 hours.Then the suspension was centrifugated at 1 500 rpm for 5 min.The supernatant,in which there were secretions of PWN,was filtered through 0.22 μm millipore and precipitated proteins in 100% of the saturation (NH 4) 2SO 4 solution.The sediments were dialysised in redistilled water and eventually dried to powder by freezing vaccum drier.The powder,which was used to detect enzymes in polyacrylamide electrophoresis gel with the dye staining methods of some enzymes,showed that there were cellulases(E.C.3.2.1.4),protease,peroxidase(E.C.3.2.1.4)and amylase(E.C.3.2.1.1) in the secretions of PWN.It is considered from the results that the enzymes mentioned above not only just the cellulase,but all of them possibly take part in the primary reaction of pine wilt disease caused by the nematode of Bursaphelenchus xylophilus.