Li Ping, Jiang Dawei, Li Bing, Dai Xinyue, Wang Yanhong, Jiang Zhou, Wang Yanxin
State Key Laboratory of Biogeology and Environmental Geology, China University of Geosciences, Wuhan, 430074, People's Republic of China.
Ecotoxicology. 2014 Dec;23(10):1878-89. doi: 10.1007/s10646-014-1316-5. Epub 2014 Aug 21.
A survey of bacterial and archaeal community structure was carried out in 10 shallow tube wells in a high arsenic groundwater system located in Hetao Basin, Inner Mongolia by 16S rRNA gene based two-step nested PCR-DGGE, clone libraries and 454 pyrosequencing. 12 bacterial and 18 archaeal DGGE bands and 26-136 species-level OTUs were detected for all the samples. 299 bacterial and 283 archaeal 16S rRNA gene clones for two typical samples were identified by phylogenetic analysis. Most of the results from these different methods were consistent with the dominant bacterial populations. But the proportions of the microbial populations were mostly different and the bacterial communities in most of these samples from pyrosequencing were both more abundant and more diverse than those from the traditional methods. Even after quality filtering, pyrosequencing revealed some populations including Alishewanella, Sulfuricurvum, Arthrobacter, Sporosarcina and Algoriphagus which were not detected with traditional techniques. The most dominant bacterial populations in these samples identified as some arsenic, iron, nitrogen and sulfur reducing and oxidizing related populations including Acinetobacter, Pseudomonas, Flavobacterium, Brevundimonas, Massilia, Planococcus, and Aquabacterium and archaeal communities Nitrosophaera and Methanosaeta. Acinetobacter and Pseudomonas were distinctly abundant in most of these samples. Methanogens were found as the dominant archeal population with three methods. From the results of traditional methods, the dominant archaeal populations apparently changed from phylum Thaumarchaeota to Euryarchaeota with the arsenic concentrations increasing. But this structure dynamic change was not revealed with pyrosequencing. Our results imply that an integrated approach combining the traditional methods and next generation sequencing approaches to characterize the microbial communities in high arsenic groundwater is recommended.
通过基于16S rRNA基因的两步巢式PCR-DGGE、克隆文库和454焦磷酸测序技术,对内蒙古河套盆地高砷地下水系统中的10口浅管井进行了细菌和古菌群落结构调查。所有样品共检测到12条细菌DGGE条带、18条古菌DGGE条带以及26 - 136个物种水平的操作分类单元(OTU)。通过系统发育分析鉴定了两个典型样品中的299个细菌16S rRNA基因克隆和283个古菌16S rRNA基因克隆。这些不同方法得到的大多数结果与优势细菌种群一致。但微生物种群的比例大多不同,且焦磷酸测序得到的大多数样品中的细菌群落比传统方法得到的更丰富、更多样。即使经过质量过滤,焦磷酸测序仍揭示了一些传统技术未检测到的种群,包括阿氏杆菌属、嗜硫弧菌属、节杆菌属、芽孢八叠球菌属和噬冷杆菌属。这些样品中最主要的细菌种群被鉴定为一些与砷、铁、氮和硫还原及氧化相关的种群,包括不动杆菌属、假单胞菌属、黄杆菌属、短波单胞菌属、马赛菌属、动性球菌属和水生杆菌属,以及古菌群落亚硝化泉古菌属和甲烷鬃菌属。不动杆菌属和假单胞菌属在大多数这些样品中明显丰富。三种方法均发现产甲烷菌是主要的古菌种群。从传统方法的结果来看,随着砷浓度的增加,优势古菌种群明显从泉古菌门变为广古菌门。但焦磷酸测序未揭示这种结构动态变化。我们的结果表明,建议采用传统方法与新一代测序方法相结合的综合方法来表征高砷地下水中的微生物群落。