摘要
土壤中分离的枯草芽胞杆菌,其发酵液对苹果炭疽病菌具有很强的抑制作用。为了确定其抑菌物质的主要成分,本试验采用硫酸铵分级沉淀获得无菌滤液中的抗菌蛋白粗提物,超滤后经过聚丙烯酰胺凝胶电泳,胶带回收进行分离鉴定,得到1种具有抑菌活性的蛋白条带。该蛋白对供试的6种植物病原真菌具有抑制作用,该蛋白的活性表现在80℃,30 min 仍有抑菌作用,对 pH（4～7）具有一定的适应范围、另外对紫外照射（0～12 h）、抑制剂、有机溶剂均不敏感。质谱鉴定该条带含有两种蛋白,分别为假定蛋白 BSU35980 gi｜16080651,相对分子量11079 Da,等电点4.78,匹配率为68%；丝氨酸羟甲基转移酶 gi｜16080743,相对分子量为45575 Da,等电点为5.56,匹配率为67%。显微观察发现抑菌蛋白可以使菌丝发生断裂、扭曲、畸形、肿大。
Bacillus subtilis was isolated from the soil,and the fermentation liquid had strong inhibitory action a-gainst Colletotrichum gloeosporioides .In order to identify the antimicrobial compounds,the antimicrobial sub-stances were isolated and purified by （NH4 ）2 SO4 fractional precipitation,ultrafiltration,PAGE-polyacrylamide gel electrophoresis,and tape reciting.The results showed that the purified active protein had inhibitory action to 6 plant pathogenic fungi.The protein was found to be stable at 80 ℃ for 30 min.The inhibitory activity of the protein was observed in the range of pH value from 4 to 7,and not sensitive to ultraviolet radiation,inhibitor and organic solvents.Two kinds of proteins contained in this stripe were identified by mass spectrum.They were the hypothetical protein BSU35980 gi｜1 608065 1,with a relative molecular mass of 1 1 079 Da and isoelectric point （p I）of 4.78,and a matching ratio of 68%,and serine hydroxymethyltransferase gi｜1 6080743,with a relative molecular mass of 45 575 Da and isoelectric point （p I）of 5.56,and a matching ratio of 67%.Microscope obser-vation revealed that the protein could break up and enlarge the mycelia.
出处
《植物保护》
CAS
CSCD
北大核心
2015年第6期49-54,共6页
Plant Protection
基金
安徽省教育厅自然科学重点科研项目(KJ2007A095)
关键词
枯草芽胞杆菌
凝胶电泳
理化性质
质谱鉴定
Bacillus subtilis
PAGE-polyacrylamide gel electrophoresis
physiochemical property
mass spectrum identification
