根肿(Plasmodiophora brassicae)休眠孢子的纯化和超声波破碎方法研究

Purification Method and Ultrasonic Disruption of Plasmodiophora brassicae Resting Spores

根肿(Plasmodiophora brassicae Woronin)是严重危害十字花科植物的世界性病害.采用蔗糖密度梯度离心和超声波处理方法从油菜染病根部组织分离和提纯根肿休眠孢子.众多因素可以影响提纯休眠孢子的纯度,为了得到更高纯度和均匀度的休眠孢子的优化方法,我们对一系列影响因素如超声波破碎功率、pH、孢子破碎浓度、处理时间和不同的水溶物质等进行了研究.在pH值5.5左右、添加0.5%的二甲基亚砜、孢子浓度约为7×108、345W功率下超声波处理10min条件下获得了最优数量和均匀度的孢子,杂菌数量减少了51%以上,杂菌种类减少超过75%.超声波处理50min后,孢子破碎率稳定地达到90%以上,除菌率达99%以上.用此方法能在较短时间内获得高纯度和高均匀度的休眠孢子,为休眠孢子的深入研究提供了资料.

Plasmodiophora brassicae Woronin is a serious worldwide disease of the plants of Cruciferae. The resting spores of P. brassicae were isolated and purified from infected root tissues of rape by density gradient centrifugation of sucrose and ultrasonic treatment. There were many factors that could effect the purification of resting spores. In order to optimize a method to obtain more purified and uniformity resting spores, various values of ultrasonic power, pH, spores concentration, treatment time and different water - solubility solutes were studied. The best uniformity of the spore suspension was obtained under the optimized conditions, pH 5.5, DMSO concentration 0.5%, spores concentration 7 × 10^8 and power value 345 W for 10 min. The miscellaneous microorganisms were reduced by more than 51%, and the species of miscellaneous microorganisms reduced by more than 75%. Almost 90% spores were disrupted by ultrasonic treatment for 50 min and over 99% miscellaneous microorganisms were removed. The purified and dispersed spores could be harvested in a short time by this method. It may provide some useful data for further studying on resting spores of P. brassicae. Fig 4, Tab 3, Ref 10