柞蚕微孢子虫孢子分离纯化方法

Study on the method of separation and purification in Nosema pernyi

柞蚕微粒子病是柞蚕Antheraea pernyi(Guérin-Méneville)的主要胚胎传染性病害,病原为柞蚕微孢子虫Nosema pernyi(Wenet Ding),其病原分离提纯技术研究对于柞蚕微粒子病的防治具有重要意义。本文利用差速离心和Percoll密度梯度离心法研究了柞蚕微孢子虫孢子的分离纯化方法,结果表明,采用不连续密度梯度分离纯化柞蚕微孢子虫孢子的效果比单一浓度的效果好,以浓度为25%、50%、75%、100%不连续梯度,15000r/min离心30min分离纯化得到的柞蚕微孢子虫孢子纯净度高。

Nosema pernyi（Wen et Ding） is a lethal pathogen caused pebrine disease in Antheraea pernyi（Guérin-Méneville）.Pebrine disease,which is vertically transmitted,is one of the most harmful diseases of A.pernyi.Better information on this disease and its vector are required to prevent outbreaks.This paper compares two methods of separating and purifying the spores of N.pernyi;differential centrifugation and Percoll density gradient centrifugation.The results show that discontinuous density gradient centrifugation was superior to single density gradient centrifugation.The purity of spores centrifuged at 15 000 r /min for 30 min under a discontinuous density gradient was 25%,50%,75% and 100%.