广西红树林根际土壤放线菌的原位培养分离及其活性筛选  

In situisolation of actinomycetes and screening bioactive potential from mangrove rhizosphere soils in Guangxi

作　　者：姜明国[1,2] 甘光华[1,2] 杨立芳[3] 李西妮[2] 杨桂柳 庹利 孙承航[4] 黄玲[3] 蓝金枝 

机构地区：[1]广西红树林保护与利用重点实验室,广西北海536000 [2]广西民族大学广西高校微生物与植物资源利用重点实验室,广西南宁530008 [3]广西民族大学化学化工学院广西林产化学与工程重点实验室,广西南宁530008 [4]中国医学科学院/北京协和医学院医药生物技术研究所,北京100050

出　　处：《海洋学报》2015年第2期55-64,共10页

基　　金：广西红树林保护重点实验室(GKLMC-201204);国家自然基金(31260004;81172963);广西自然基金(2012GXNSFAA053038);高等学校博士学科点专项科研基金博导类项目(20111106110032);广西民族大学重点科研项目(2012MDZD041)

摘　　要：为了发掘广西红树林根际土壤的放线菌资源,本文利用原位培养装置,埋于根际土壤中俘获放线菌,30d后取回实验室,采用平板涂布法对4个地点的原位培养样品于15种培养基上进行分离纯化;对分离株基于16SrRNA基因序列进行系统发育分析;进一步进行抗菌活性和产酶活性检测。共分离得到113株放线菌。对其中33株放线菌进行测序,结果表明20株属于链霉菌属,11株属于拟诺卡氏菌属,1株属于伦兹氏菌属,1株与拟诺卡氏菌属相似性最高为90%,很可能属于放线菌一个新属。抗菌实验结果显示其中有7株、4株、18株、6株、10株、3株实验菌株分别对大肠埃希菌、铜绿假单胞菌、金黄色葡萄球菌、普通变形杆菌、乙型溶血性链球菌和肺炎克雷伯氏菌具有抑制作用;有55株、62株、24株、72株的实验菌株分别具有纤维素酶活性、淀粉酶活性、胶原蛋白酶活性、酯酶活性。原位培养可以丰富对广西红树林根际土壤的认识,分离到了新种甚至可能是新属的放线菌,分离得到的部分放线菌菌株具有较高生物活性,为后续工作提供了良好的实验材料。In order to efficiently capture actinomycetes from mangrove rhizosphere soils in Guangxi,traps for particularly cultivating actinomycetes in situ were implanted into soil and were incubated for thirty days.A general selective isolation procedure was employed onto fifteen different media.Phylogenetic neighbour-joining tree was conducted on the 16 SrRNA gene sequences of the isolates and type strains.The biological activities and enzyme production of the isolated strains were evaluated.One hundred thirty-three actinomycetes were recovered.Of the thirtythree randomly picked isolates,twenty were Streptomyces,eleven were Nocardiopsis and one was Actinosynnema.One isolate,GXUN10,was most closely related to Nocardiopsis with a low 16 SrRNA gene sequence identity of90.0%,and might represent a new genus.Seven,four,eighteen,six,ten and three isolates showed antimicrobial activities respectively to Escherichia coli,Pseudomonas aeruginosa,Staphylococcus aureus,Proteusbacillus vulgaris,Hemolytic streptococcus and Pneumococcus.Fifty-five,sixty-two,twenty-two and seventy-two isolates respectively showed cellulase,amylase,collagen enzyme and esterase activities.This study provides further evidence that unusual and rare actinomycetes from mangrove rhizosphere soils in Guangxi can be recovered by in situcultivation traps.

关 键 词：原位培养 红树林 根际土壤 放线菌 生物活性 

分 类 号：Q938[生物学—微生物学]