玻利维亚进口盐中可培养嗜盐古菌组成分析  

Culturable Halophilic Archaea in the Commercial Salts Imported From Bolivia

作　　者：刘靖文 刘柳 陈飞龙 陈绍兴 LIU Jing-wen;LIU Liu;CHEN Fei-long;CHEN Shao-xing(College of Life Sciences,Anhui Normal University,Wuhu 241000,China)

机构地区：[1]安徽师范大学生命科学学院,安徽芜湖241000

出　　处：《安徽师范大学学报（自然科学版）》2021年第3期250-258,共9页Journal of Anhui Normal University(Natural Science)

基　　金：国家自然科学基金项目(31460003);安徽省重要生物资源保护与利用开放课题(591601).

摘　　要：嗜盐古菌是一类通常生活于高盐环境的极端微生物,独特的细胞结构和生理代谢系统赋予其重要的研究价值。本研究通过获取玻利维亚进口商品盐中嗜盐古菌资源,筛选一批具有较高胞外酶活、产嗜盐菌素的嗜盐古菌,并研究该生境中嗜盐古菌的群落组成。本研究采用不同pH值的嗜盐古菌培养基,以纯培养划线分离的方式,结合16S rRNA基因的测序和序列相似性分析,获得所分离菌株的分类地位并进行物种多样性分析。此外,采用胞外酶活的检测方法,对部分菌株的胞外酶酶活和产嗜盐菌素能力进行筛查。最终从该样品盐中共分离到505株嗜盐古菌,分别来自于Natronococcus、Natrinema、Natrialba、Halovivax、Haloterrigena、Halorubrum、Halopenitus、Halobellus、Halococcus、Halobacterium、Halalkalicoccus、Haloarcula和Halosimplex等13个属,其中有180株属于Halobacterium属,占比约35.6%,是本研究纯培养分离所揭示的优势类群。物种多样性分析结果显示该样品中可培养嗜盐古菌数量较高、物种数目多,但优势种群明显且均匀度不高,总体而言多样性不高。不同pH培养基纯培养分离结果显示:pH8.0和pH9.0的培养条件下分离得到的菌株物种多样性较高:前者Shannon指数为2.8,simpson指数为0.9;后者Shannon指数为2.4,simpson指数为0.9。胞外酶活检测发现6株产胞外淀粉酶,5株产胞外明胶酶,2株产胞外蛋白酶菌株,胞外氧化酶和触酶活性在嗜盐古菌中较为普遍。拮抗作用检测获得1株产嗜盐菌素的菌株。本研究通过纯培养方法获得了丰富的嗜盐古菌菌株资源,并初步揭示了玻利维亚进口商品盐样品中可培养嗜盐古菌的群落组成和多样性分布。Halophilic archaea is an extremophile which usually lives in a hypersaline environment.Its unique cell structure and metabolic system endow it an important research value.In order to investigate the species diversity and obtain some extracellular enzymes and halocin producing strains from world-wide hypersaline environment,a commercial salt sample imported from Bolivia was studied.In this study,medium with high salt concentration under different pH values was used,and pure cultures were isolated via continuous streaking.The 16S rRNA gene was sequenced with universal pair of primers.Based on sequence similarity search of the 16S rRNA gene,taxonomic position of each strain and the profile of the species diversity under different pH conditions were obtained.In addition,extracellular enzyme activities were detected on agar plate with chemical regents.Halocin activity was screen with four different indicators on agar plate.Finally,A total of 505 halophilic archaeal strains belonging to 13 genera,e.g.,Natronococcus,Natrinema,Natrialba,Halovivax,Haloterrigena,Halorubrum,Halopenitus,Halobellus,Halococcus,Halobacterium,Halkalicoccus,Haloarcula and Halosimplex,were isolated.Among them,180 strains belonged to Halobacterium,accounting for about 35.6%,which was the dominant group.Culturable species of halophilic archaea are rich in this sample,but the dominant group is obvious because of the low evenness index.Strain isolation under different pH media,we found that the species diversity index under pH 8.0(Shannon index:2.8;Simpson index:0.9)was higher than that of pH 9.0(Shannon index:2.4;Simpson index:0.9).Also we isolated 6 strains with extracellular amylase activities,5 strains with extracellular gelatinase activities,and 2 strains with extracellular protease activities.Activities of extracellular oxidase and catalase were common in halophilic archaea.A halocin-producing strain was obtained.In this study,pure culture approach revealed that there were abundant resources of halophilic archaea in this sample,and species diver

关 键 词：极端环境微生物 高盐环境 嗜盐古菌 16S rRNA基因 物种多样性 

分 类 号：Q93[生物学—微生物学]