机构地区: 华南农业大学资源环境学院植物病理学系
出 处: 《华中农业大学学报》 2007年第1期37-40,共4页
摘 要: 利用蛋白纯化系统AKTA Purifier 100对来源于稻瘟菌(Magnaporthe grisea)ZC13菌株97-151a菌丝细胞壁的糖蛋白激发子CSBⅠ进行纯化过程的优化。离子交换层析条件的选择结果表明:弱阴离子柱DE-AE-Sepharose FF和Tris-HCl缓冲体系的洗脱效果较好。新的步骤是:选用YM30膜超滤,省略ConA-Sepha-rose 4B亲和层析,粗激发子过凝胶柱Sephacryl S-100,阴离子柱DEAE-Sepharose FF,获得的活性峰D3,即为纯化的CSBⅠ。随着激发子的逐步纯化,其诱导不同水稻品系叶片中的POD比活相应显著升高(P<0.01),CSBⅠ的诱导活性最高。该纯化方法简便,重现性稳定,提高了纯化效率。 A procedure for purifying a glycoprotein elicitor CSB I from hyphal cell walls of the strain 97-151a of M. grisea race ZC13 equipped with AKTA Purifier 100 system was optimized. After centrifugation, uhrafihration with YM30 membrane, crude elicitor fraction was applied to Sephacryl S-100 column, then the active peak S1 was injected onto DEAE-Sepharose fast flow column. The bioassay indicated that elicitor activity was restricted to the symmetric elution peak D3. It was named CSB Ⅰ. CSB Ⅰ showed as a single band on SDS-PAGE followed by Commassie Brilliant Blue R-250 staining or periodic acid-Schiff staining. With the further purification, the induction of the peroxidase activity in different rice cuhivar leaves after treatment with each step elicitor was higher (P〈0. 01). The new effective strategy should provide useful information on discussing the possible implication of such a glycoprotein elicitor in plant-microbe interactions.