塔里木河流域土壤小单孢菌的选择性分离及其抗菌活性筛选  

Selective isolation and antibacterial screening of Micromonospora from soils in Tarim River Basin

作　　者：梁红丽 沈红玲[1] 杨秋红 万传星[1] LIANG Hongli;SHEN Hongling;YANG Qiuhong;WAN Chuanxing(College of Life Science and Technology,Tarim University/Xinjiang Production&Construction Crops Key Laboratory of Protection and Utilization of Biological Resources in Tarim Basin.Alar,Xinjiang 843300)

机构地区：[1]塔里木大学生命科学与技术学院/塔里木盆地生物资源保护利用兵团重点实验室,新疆阿拉尔843300

出　　处：《塔里木大学学报》2023年第1期19-30,共12页Journal of Tarim University

基　　金：新疆维吾尔自治区研究生科研创新项目“小单孢菌的分离鉴定与氨基糖苷类次生代谢产物的挖掘”(XJ2022G239);塔里木大学校长基金硕士项目“基因组挖矿技术指导下的新糖肽类抗生素发现研究”(TDZKSS202001)。

摘　　要：小单孢菌(Micromonospora)是产抗最多的稀有放线菌,为了探索塔里木河流域土样中小单孢菌物种多样性并筛选抗菌活性菌株,本研究通过可培养的方法以塔里木河流域采集的10份土样为研究对象,采用6种不同的分离培养基,通过添加特定抗生素选择性分离培养小单孢菌,并对小单孢菌进行物种多样性及次生代谢产物生物合成基因簇分析。结果表明:选用可培养方法从塔里木河沿岸采集的土样中分离鉴定出484株菌株,分属于3门5纲13目19科22属70种,其中小单孢菌属菌株424株,占87.60%,小单孢菌属的出菌率高。以金黄色葡萄球菌、大肠杆菌、鼠伤寒沙门氏菌、福氏志贺氏菌、解淀粉欧文氏菌和白色念珠菌等病原菌作为靶标菌对小单孢菌的发酵粗提物进行抗菌活性筛选,结合antiSMASH生物合成基因簇分析,筛选出5株既具有抗菌活性又具有代谢潜力的小单孢菌。研究表明塔里木河沿岸含水量高的土样中拥有丰富的小单孢菌资源,通过向分离培养基中添加新生霉素、萘啶酮酸、放线菌素等抗生素施加选择压,可以有选择性地分离获得小单孢菌。Micromonospora produced most of the antibiotics among rare actinomycete strains.In order to explore the species diversity of Micromonospora in the soil of the Tarim River Basin and screen the strains with antibacterial activity,10 soil samples collected from the Tarim River Basin.Micromonospora were selectively isolated and cultured by adding specific antibiotics in 6 different isolation media.The species diversity and biosynthesis gene clusters of secondary metabolites in Micromonospora were analyzed.The results showed that 484 strains belonged to 70 species,22 genera,19 families,13 orders,5 classes,and 3 phyla were isolated and identified,among which included 424 strains of Micromonospora,accounting for 87.60%.The effect of selective isolation was obvious.Staphylococcus aureus,Escherichia coli,Salmonella typhimurium,Shigella flexneri,Erwinia amylovora,and Candida albi-cans were selected as target pathogenic bacteria to evaluate the antibacterial activity of the fermentation crude extract of Micro-monospora.Combined with antiSMASH biosynthesis gene cluster analyses,5 Micromonospora strains with both antibacterial activity and metabolic potential were screened.The research showed that there were abundant Micromonospora resources in the wet soil in the Tarim River Basin.By adding neomycin,naphthylpyruvic acid,actinomycin and other antibiotics into the isolation media and applying selective pressure,Micromonospora can be directionally isolated.

关 键 词：塔里木河流域 小单孢菌 选择性分离 多样性 抗菌活性 

分 类 号：Q939.[生物学—微生物学]