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新疆罗布泊盐湖放线菌多样性及多相分类
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摘要
嗜盐放线菌是一类存在于天然高盐域中的极端微生物,起源早,生存环境恶劣。作为一种生命的奇迹,其特殊的生理机制,基因类型和代谢产物成为认识生命现象,阐明生物多样性形成机制和推动生物技术产业健康快速发展的宝贵资源,也是各国争夺战略资源和相关高新技术知识产权的一个重要领域。
     至今为止,人类获得的纯培养微生物仅为实有微生物数量的1%,极端微生物更是稀少。物种资源的严重匮乏无疑限制了微生物资源开发的深度和广度。为此,笔者以罗布泊盐湖这个高温、干旱、高盐、贫瘠的极端生态环境为对象,通过非培养和可培养手段研究极端环境放线菌资源,客观认识盐湖生态放线菌的多样性,加深人们对宇宙生命现象的理解,以及由此而产生的可能对生命科学发展带来的深远意义。
     关于罗布泊盐湖非培养放线菌的研究,一些令人惊喜的结果被发现。首先,罗布泊盐湖放线菌多样性极为丰富,Ilumatobacter, Saccharopolyspora, Streptomyces, Actinopolyspora, Kocuria, Streptomonospora, Streptosporangium, Amycolatopsis, Isoptericola, Nocardiopsis, Dietzia和Glycomyces共12个属的放线菌被获得;其次,盐湖中存在有大量放线菌新类群,58.82%克隆的序列同源性≤97%,36.78%的序列无法归入任何一个属,预示着可能是一些科一级或更高的新的放线菌分类单元;最后,放线菌群落中占优势的菌群为Glycomyces (20.1%), Dietzia (16.13%), Isoptericola (10.3%)和Nocardiopsis (5.88%).同时,Saccharopolyspora和Actinopolyspora的比例最小,仅为0.49%。最值得注意的一点是普通土壤环境广泛生存的优势种链霉菌Streptomyces在罗布泊这个极端干旱高盐的环境中仅占到1.47%。说明盐湖这种生态环境中更容易获得稀有放线菌,是挖掘放线菌新物种资源的理想之地。
     可培养的结果更让我们惊叹于微生物强大的生存与繁殖能力。共有18个属的放线菌被发现,即嗜盐糖霉菌属(Haloglycomyces)、链霉菌属(Streptomyces)、放线多孢菌属(Actinopolyspora)、糖霉菌属(Glycomyces)、小单孢菌属(Micromonospora)、链单孢菌属(Streptomonospora)、糖单孢菌属(Saccharomonospora)、拟无枝菌酸菌属(Amycolatopsis)、拟诺卡氏菌属(Nocardiopsis)、产丝菌属(Myceligenerans)、考克氏菌属(Kocuria)、栖白蚁菌属(Isoptericola)、普劳斯氏菌属(Prauserella)、短杆菌属(Brevibacterium)、糖多孢放线菌属(Saccharopolyspora)、疣孢菌属(Verrucosispora)、涅斯捷连科氏菌属(Nesterenkonia)和嗜盐放线产孢菌属(Haloactinospora)。发现新物种8个,其中6个新种通过多相分类技术被确定其系统分类地位。菌株YIM92370的16SrRNA基因序列相似性与亲缘关系最近的同源菌株Glycomyces arizonensis仅有93.7%,定为糖霉菌科的1个新属,即嗜盐糖霉菌属(Haloglycomyces),白色嗜盐糖霉菌新种(Haloglycomyces albus);菌株TRM415是Brevibacterium属的一个新成员,命名为盐生耐盐短杆菌(Brevibacterium salitolerans);菌株TRM40136是Actinopolyspora属内的一个新分类单元。命名为新疆放线多孢菌(Actinopolyspora xinjiangensis);菌株TRM40133是糖多孢菌属的一个新成员,分离至罗布泊盐湖,因此,命名为盐湖糖多孢菌(Saccharopolyspora lacisalsi);菌株TRM40137为糖霉菌属的一个新成员,命名为喜盐耐盐糖霉菌(Glycomyces halotolerans);菌株TRM F103是Amycolatopsis属的新成员,命名为盐生耐盐拟无枝菌酸(Amycolatopsis salitolerans)。就实验结果而言,GFA、GW1、AGG、 MM和GL3培养基得到的放线菌多样性及其新类群较多,可考虑用于盐湖生态放线菌的分离培养基。由于不同放线菌对盐浓度依赖性的不同,根据不同盐浓度放线菌的分离结果,建议分离盐湖放线菌时采用多个盐浓度。总的来说,世界上没有万能的培养基,也没有统一的分离条件,只有不断的设计和更新分离手段,才能更好的反映真实的微生物世界。
Halophilic actinomycetes from extremely hypersaline environments are an age-old microorganism. Therefore, they could adapt to the peculiar environment conditions and produce some unique gene types and natural active compounds. Extreme environmental microbe is a valuable source for knowing the mechanism of biodiversity and boosting the development of biotechnology industry. Extremophiles are some strategic biosources for the protection of intellectual property rights in novel technology field.
     To this day, culture-dependent microbe quantity, especially extremophiles are very little, which seriously restrict the exploitation and utilization of microbial source. Phylogenetic diversity of halophilic actinomycetes from hypersaline, arid and sterile environments in Lop Nur salt lake was investigated by using the culture-dependent and culture-independent methods and phylogenetic analysis based on16S rRNA gene sequences. So as to we impersonally know actinobacterial diversity and to deeply understand life phenomenon.
     As far as the result of culture-independent methods on actinobacterial diversity, some good results are detected in this study. Firstly, The results showed that there were abundant actinobacterial diversity in the sediment of Lop Nur salt lake. These clones belong to12genera in the Acidimicrobidae, namely Ilumatobacter, Saccharopolyspora, Streptomyces, Actinopolyspora, Kocuria, Streptomonospora, Streptosporangium, Amycolatopsis, Isoptericola, Nocardiopsis, Dietzia and Glycomyces. Secondly, abundant novel actinobacteria exit in the salt lake.58.82%clones sequences exhibited low similarity (<97.0%) to other species of the Acidimicrobidae which represent novel actinobacteria taxa. Others (36.78%) exhibited lower similarity (<92.0%) which formed some large distinct clades in phylogenetic tree of actinobacterial16S rRNA gene sequences among Phylum Actinobacteria, may represent some new suborder or new class. Finally, the most abundant and diverse clones were the Glycomyces (20.1%), Dietzia(16.13%), lsoptericola(10.3%) and Nocardiopsis(5.88%), not Streptomyces(1.47%). The results showed that there were abundant actinobacterial diversity and some unknown actionobacterial groups existed in hypersaline environments in the sediment of Lop Nur salt lake.
     These actinomycetes isolated from hypersaline Environments belong to18genera, namely Haloglycomyces, Streptomyces, Actinopolyspora, Glycomyces, Micromonospora, Streptomonospora, Saccharomonospora, Amycolatopsis, Nocardiopsis, Myceligenerans, Kocuria, Isoptericola, Prauserella, Brevibacterium, Saccharopolyspora, Verrucosispora, Nesterenkonia and Haloactinospora. Excitingly, eight novel strains ware isolated. In accordance with the principle of polyphasic taxonomy, a systematic study had been conducted on the taxonomy of six novel isolates. Phylogenetic analysis showed that strain YIM92370T, having lower16S rRNA gene sequence similarity of93.7%with the described species of this Glycomycetaceae, can be distinguished from representatives of other two genera Glycomyces and Stackebrandtia in the family. The novel genus and species Haloglycomyces albus gen. nov., sp. nov. were proposed, with strain YIM92370Tas the type strain of Haloglycomyces albus. On the basis of the polyphasic evidence, the strain TRM40136T should be designated as a novel species of the genus Actinopolyspora for which the name Actinopolyspora xinjiangensis sp. nov. In addition, three novel halotolerans actinobacteria species(Brevibacterium salitolerans, Glycomyces halotolerans and Amycolatopsis salitolerans) as well as one holaphilic species(Saccharopolyspora lacisalsi) were proposed.
     According to test result, five media (GFA、GW1、AGG、MM and GL3) can good to isolate actinobacteria of salt lake. But no medium is almighty. Accordingly, we should design and innovate more media or isolated methods to make better to know the life world.
引文
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