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16S rRNA高通量测序研究集雨窖水中微生物群落结构及多样性
摘要点击 5631  全文点击 1514  投稿时间:2016-08-11  修订日期:2016-11-10
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中文关键词  典型集雨饮水地区  集雨窖水  Illumina MiSeq高通量测序  细菌多样性  潜在致病菌
英文关键词  typical rainwater harvesting area  cellar water  MiSeq high-throughput sequencing  bacterial diversity  potential pathogen
作者单位E-mail
杨浩 兰州交通大学环境与市政工程学院, 兰州 730070
寒旱地区水资源综合利用教育部工程研究中心, 兰州 730070 
haoyang7614236@163.com 
张国珍 兰州交通大学环境与市政工程学院, 兰州 730070
寒旱地区水资源综合利用教育部工程研究中心, 兰州 730070 
 
杨晓妮 甘肃农业大学资源与环境学院, 兰州 730070  
武福平 兰州交通大学环境与市政工程学院, 兰州 730070
寒旱地区水资源综合利用教育部工程研究中心, 兰州 730070 
 
赵炜 兰州交通大学环境与市政工程学院, 兰州 730070
寒旱地区水资源综合利用教育部工程研究中心, 兰州 730070 
 
张洪伟 兰州交通大学环境与市政工程学院, 兰州 730070  
张翔 兰州交通大学环境与市政工程学院, 兰州 730070  
中文摘要
      为探究集雨窖水中细菌多样性及功能与主要水质因子之间关系,应用16S rRNA基因-Illumina MiSeq高通量测序技术分析和比较了不同类型集雨水窖中窖水的微生物群落结构及多样性.通过提取4组样品基因组DNA,对16S rRNA V4区测序,共获得用于物种分类的OTU 1605条,涵盖了22门、42纲、71目、115科、146属的细菌.微生物多样性分析表明,窖水具有很高的细菌多样性,不同样品多样性存在差异,细菌优势菌群分布呈现出大量的稀有种和少数常见种的模式.菌群分类发现,拟杆菌门(Bacteroidetes)、变形菌门(Proteobacteria)、放线细菌门(Actinobacteria)、疣微菌门(Verrucomicrobia)和OD1是主要的细菌菌门,其相对丰度占群落的87.1%~94.8%;放线菌门的放线细菌纲(Actinobacteria)、酸微菌纲(Acidimicrobiia),拟杆菌门的噬纤维菌纲(Cytophagia)、黄杆菌纲(Flavobacteriia)、鞘脂杆菌纲(Sphingobacteriia),变形菌门中α-变形菌纲(α-Proteobacteria)、β-变形菌纲(β-Proteobacteria)、γ-变形菌纲(γ-Proteobacteria),疣微菌门丰收神菌纲(Opitutae)、疣微菌纲(Verrucomicrobiae)、Pedosphaerae,OD1的ZB2为纲层次上的优势菌;红细菌属(Rhodobacter)、脱氮单孢菌属(Dechloromonas)、黄杆菌属(Flavobacterium)、不动杆菌属(Acinetobacter)、丛毛单孢菌属(Comamonas)、假单孢菌属(Pseudomonas)、嗜氢菌属(Hydrogenophaga)等优势菌属大多数为具有脱氮除磷功能的反硝化菌和异养硝化-好氧反硝化菌;RDA分析表明,不同环境因子对微生物群落的影响有着明显的不同,即第Ⅱ簇内的菌属主要与反映窖水有机物含量的指标UV254、高锰酸盐指数、BOD5之间呈显著正相关关系,而第Ⅲ簇内的菌属主要与反映窖水氮、磷水平的指标TN、NO2--N、NO3--N、TP、NH4+-N显著正相关,体现了生态功能的簇内相似性及簇间差异性.研究结果加深了对窖水微生物群落结构和多样性的认识,为深入研究窖水细菌多样性及功能与水质因子之间关系提供了借鉴.
英文摘要
      This study was aimed to explore the bacterial diversity of cellar water as well as to study the relationship between the bacterial diversity and environmental factors. The MiSeq high-throughput sequencing was used to analyze and compare the bacterial diversity and community composition of samples from different cellar water samples. Overall 1605 optimized reads were obtained from four samples based on high-throughput sequencing of the V4 region of the 16S rRNA gene. Bacterial species detected in these samples covered 22 phyla,42 classes,71 orders,115 families, 146 genera. Analysis showed that the bacterial diversity was very high in these samples, and there were differences among different samples. The distribution characteristics of the dominant bacteria showed patterns of a large number of rare species and a few common types. Taxonomic assignment analysis indicated that Bacteroidetes,Proteobacteria,Actinobacteria,Verrucomicrobia,OD1 dominated in the Cellar water, and accounted for 87.1% to 94.8% at phylum level. The predominant groups were Actinobacteria,Acidimicrobiia,Cytophagia, Flavobacteriia, Sphingobacteriia,α-Proteobacteria,β-Proteobacteria,γ-Proteobacteria,Opitutae, Verrucomicrobiae,Pedosphaerae and ZB2 at class level. At genus level Rhodobacter,Dechloromonas,Flavobacterium,Acinetobacter,Comamonas,Pseudomonas,Hydrogenophaga,et al were the abundant taxa, which were mainly denitrifying bacteria and heterotrophic nitrification-aerobic denitrification bacteria. The result of RDA suggested that the influences of different environmental factors on different microbes were different. Bacterial community Ⅱ had significant positive correlation with UV254,permanganate index,BOD5,and Bacterial community Ⅲ had significant positive correlation with TN,NO2--N,NO3--N,TP,NH4+-N. This research should deepen the understanding on microbial community in Cellar water, and provide references for the association of bacterial composition and diversity with environmental factors.

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