高通量测序分析云南腾冲热海热泉微生物多样性

Amplicon-based high-throughput sequencing reveals the microbial diversity in Rehai hot springs, Tengchong, Yunnan Province

【背景】云南腾冲热海热泉中蕴含着丰富的极端微生物资源。【目的】揭示云南腾冲热海热泉中微生物物种多样性及群落结构差异,发掘酸性热泉中铁、硫氧化功能微生物。【方法】采用Illumina HiSeq高通量测序技术对3处热泉15个水体样品中微生物16SrRNA基因V4-V5区进行测序及生物信息学分析。【结果】3处热泉中共获得578061条有效序列,聚类为141个可操作分类单元(Operational taxonomic unit,OTU),包括19个门66个属。鼓鸣泉(GMQ)、蛤蟆嘴(HMZ)、黄瓜箐(HGQ)3处热泉均以泉古菌门(Crenarchaeota)和厚壁菌门(Firmicute)为主。从属水平分析,碱性热泉鼓鸣泉(GMQ)和中性热泉蛤蟆嘴(HMZ)分别注释到37、32个属,优势属均为芽孢杆菌属(Bacillus)和热棒菌属(Pyrobaculum)。酸性热泉黄瓜箐(HGQ)共注释到20个属,优势属为酸杆菌属(Acidibacillus)和酸硫杆状菌属(Acidithiobacillus),此外,具有铁、硫氧化潜力的菌属有喜酸菌属(Acidicaldus)、硫化芽孢杆菌属(Sulfobacillus)、硫化叶菌属(Sulfolobus)及生金球菌属(Metallosphaera)等,进一步通过硫氧化培养基分离获得了这些菌属中的纯菌株。【结论】云南腾冲热海热泉水体中蕴含丰富的微生物资源,热泉间微生物物种组成差异明显;酸性热泉中存在多种具有潜在铁、硫代谢功能的菌种;未分类类群、非培养类群丰度很高,尤其是蕴藏着可观的古菌资源。

[Background]Rehai hot springs in Yunnan Province contain a wealth of extreme microbial resources.[Objective]This study was proposed to study the microbial diversity and community structures in Rehai hot springs,especially the diversity of iron-and sulfur-oxidizing microorganisms in the acid hot springs.[Methods]The microbial 16 S rRNA V4-V5 genes region was sequenced by Illumina HiSeq technology and analyzed by bioinformatics software.[Results]A total of 578 061 high-quality sequences were obtained from 15 samples of 3 hot springs,resulting in 141 OTU belonging to 19 phyla,66 genera.Crenarchaeota and Firmicutes were main phyla in Gumingquan(GMQ),Hamazui(HMZ)and Huangguaqing(HGQ).On the genera level,there were 37 and 32 genera in alkaline hot spring GMQ and neutral hot spring HMZ,respectively,in which the major genera were Bacillus and Pyrobaculum.Among the 20 genera present in acidic hot spring HGQ,the genera Acidibacillus and Acidithiobacillus were dominant.Furthermore,some species of Acidicaldus,Sulfobacillus,Sulfolobus and Metallosphaera genera were isolated from HGQ,which have potential to oxidize iron and sulfur.[Conclusion]The microbial resources were diverse in Rehai hot springs and the microbial compositions of the three hot springs were obviously different.Iron-and sulfur-oxidizing microorganisms,uncultured and unclassified microbes,especially archaea,were rich in acidic HGQ.