Phylogenetic Analysis of Actinomycete Isolates from Iron Mine Tailings

铁矿尾矿放线菌分离株的系统发育分析(摘要)(英文)

[Objective]The aim was to discuss the actinomycete biodiversity of iron-mine tailings by phylogenetic analysis of 12 typical isolates. [Method]The genomic DNAs were extracted by phenol-chloroform method; phylogeny analysis was carried out based on 16S rDNA PCR amplification and sequencing. [Result]The results showed that all the 12 strains belong to the genus Streptomyces sharing 98.7%-99.9% similarities with their nearest known neighbors. [Conclusion]Streptomyces is the dominant culturable actinomycete group of iron mine tailings,in which there are many potential novel species.

[目的]通过确定12株典型分离株的系统发育地位,初步探讨铁矿尾矿中放线菌生物多样性。[方法]12株典型放线菌来源:将新鲜斜面菌株接种于贝奈特液体培养基中,28℃、180r/min摇床培养至对数生长中期。镜检验纯后4℃、10000r/min离心5min,收集菌体。采用酚-氯仿法提取基因组DNA,PCR扩增16SrDNA。所测序列采用Blast与GenBank中序列比较后,挑取相似性较大的菌株序列,采用MEGA4.1构建所测序列与相关种属的邻接法(Neighborjoining)系统发育树,并分析其系统发育地位。[结果]12株铁矿尾矿放线菌均属于链霉菌属(Streptomyces),其中菌株034005与红灰链霉菌(S.rubrogriseus)最近,序列相似性为99.2%;菌株035035与灰白链霉菌(S.griseoloalbus)最近,序列相似性为98.7%;菌株035015和035030与脆弱链霉菌(S.fragilis)最近,序列相似性分别为99.9%和99.6%;菌株035024与昌帕瓦特链霉菌(S.champavatii)最近,序列相似性为99.7%;菌株035004和035034均与娄彻氏链霉菌(S.rochei)最近,序列相同;菌株032004、032008和032015均与弗氏链霉菌(S.fradiae)最近,序列相似性分别为99.9%、99.8%和99.8%;菌株033003与黄褐链霉菌(S.flavofuscus)最近,序列相似性为100.0%;菌株035033与浅玫瑰链霉菌(S.roseolus)最近,序列相似性为99.8%。[结论]铁矿尾矿中可培养放线菌类群以链霉菌为主,其中存在大量潜在的新种。