解淀粉芽胞杆菌BaX030的分离鉴定及抗菌功能  

Screening and antibacterial function of Bacillus amyloliquefaciens X030

作　　者：何浩[1] 朱颖龄 迟立庆 赵子昭 王婷[1] 左明星[1] 张通[1] 周逢娟 夏立秋[1] 丁学知[1] 

机构地区：[1]湖南师范大学生命科学学院微生物分子生物学国家重点实验室培育基地,湖南长沙410081

出　　处：《微生物学报》2015年第9期1133-1143,共11页Acta Microbiologica Sinica

基　　金：国家自然科学基金(31370116);国家"863计划"(2011AA10A203);湖南省高校创新平台开放基金项目(13CY002;10CY013);湖南省生物发育工程及新产品研发协同创新中心项目(20134486);国家级大学生创新训练项目(201410542009)~~

摘　　要：【目的】芽胞杆菌是微生物活性物的重要来源,从全国各地采集的72份土壤样品中分离出339株芽胞杆菌,研究各菌株抑菌活性,分离纯化抑菌活性物,为丰富芽胞杆菌菌种资源和微生物次级代谢物的挖掘奠定实际应用基础。【方法】采用水浴加热和稀释平板涂布等方法从河南花生地采集的土壤中筛选得到一株具有很强抑菌活性的芽胞杆菌,结合形态观察、生理生化特征和16S rRNA基因序列同源性比对分析,对该菌株进行鉴定。丙酮沉淀、葡聚糖凝胶柱层析、C18反相柱层析得到Bacillus amyloliquefaciens X030抑菌活性物,LC-MS/MS鉴定其分子量。利用滤纸片扩散法和平板对峙培养法测定抑菌谱及拮抗性质。【结果】筛选分离得到一株解淀粉芽胞杆菌,归类并命名为解淀粉芽胞杆菌Bacillus amyloliquefaciens X030。Ba X030对金黄色葡萄球菌(Staphylococcus aureus)、白色念珠菌(Candida albicans)、酵母菌(Saccharomycetes)有较强抑制效果,对水稻稻瘟病菌(Pyriculariaoryzae)、辣椒尖胞炭疽病菌(Chili pointed cell anthrax)、枇杷炭疽病菌(Gloeosporium eriobotryae speg)、烟草黑胫病菌(Phytophthora parasitica)有良好拮抗活性。初步确定Ba X030产生的抑菌活性物为多肽类化合物。【结论】分离得到的B.amyloliquefaciens X030产生了一个对病原细菌具有较强抑制作用的多肽,同时该菌株在拮抗植物病原真菌方面也有明显的效果。[ Objective] We isolated 339 bacillus strains from 72 soil samples all over the country, then purified their antimicrobial compounds and studied the antibacterial activity, to enrich bacillus resources and explore their second metabolites. [ Methods] A bacillus strain with strong antibacterial activity was selected by dilution plate and water bath heating from a soil sample from a peanut plantation in Henan Province; this strain was identified according to morphological observation, physiological and biochemical characteristics, and consequences of 16S rRNA homologous analysis. Antibacterial compound from the identified strain, Bacillus amyloliquefaciens X030, was separated and purified by acetone precipitation, Sephadex chromatography, C18 reverse phase column chromatography. Its molecular weight was analyzed by LC-MS/MS. The antibacterial activity was characterized by disc diffusion and plate two-way cultivation. [ Results] Bacillus amyloliquefaciens was isolated that not only has antibacterial activity against Staphylococcus aureus, Candida albican and Saccharomycetes; but also against Pyriculariaoryzae, Chili pointed cell anthrax, Gloeosporium eriobotryae speg and Phytophthora parasitica. The compound was confirmed as polypeptide. [ Conclusion ] Bacillus amyloliquefaciens X030 can produce a polypeptide that inhibits pathogenic bacteria and plant pathogenic fungi.

关 键 词：解淀粉芽胞杆菌 抑菌 分离纯化 多肽 

分 类 号：Q93[生物学—微生物学]