对健康和患病蜜蜂幼虫血淋巴进行SDS-PAGE电泳和蛋白酶、酯酶的活性染色,以研究蜜蜂球囊菌分泌的胞外酶种类及对蜜蜂侵染的作用.结果表明:健康蜜蜂幼虫血淋巴中的蛋白含量丰富,主要由4种高分子质量的蛋白组成,而患病幼虫血淋巴中的蛋白含量很少,主要蛋白组分被降解;在患病幼虫血淋巴中可以检测到健康幼虫所没有的多种蛋白酶和酯酶的活性.可见,蜜蜂球囊菌在侵染蜜蜂幼虫的过程中涉及到胞外酶的作用,这些胞外酶参与降解蜜蜂的组织,为蜜蜂球囊菌的生长提供营养.%Bee chalk brood disease is one of the primary honeybee disease. It is caused by a entopathogenic fungi, Ascosphaera apis. Extracellular enzymes of entopathogenic fungi are often involved in host infecting process and therefore are vital pathogenic factors. Which extracellular enzymes were secreted by A. apis to infect honeybee is not clear so far. Here we collect the hemolymph from the healthy bee larvae and the A. apis infected larvae for SDS-PAGE, protease and esterase zymogram analysis. The result showed that the hemolymph protein is high content and composed of four main high molecular weight proteins in healthy bee larvae,and it is low content and composed of low molecular weight proteins in A. apis infected larvae. Multiple protease and esterase were identified from the A. apis infected larvae. Such results showed that A. apis could secreted multiple extraeellular enzymes to degrade bee larvae tissue for its growth. This is the first report that certificated the extraeellular enzymes of A. apis involving in infecting honeybee larvae, which is significant to illuminate the infecting process and pathogenic mechnism of A. apis.
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