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Assessment of the Diversity Abundance and Ecological Distribution of Members of Candidate Division SR1 Reveals a High Level of Phylogenetic Diversity but Limited Morphotypic Diversity

机译:SR1候选成员的多样性丰度和生态分布的评估显示出高水平的系统发育多样性但有限的形态型多样性

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摘要

We used a combination of 16S rRNA gene clone library surveys, quantitative PCR (qPCR) analysis, and fluorescent in situ hybridization to investigate the diversity, abundance, and distribution of members of candidate division SR1 in multiple habitats. Using SR1-specific 16S rRNA gene primers, we identified multiple novel SR1 lineages in four different anaerobic environments: sediments from Zodletone Spring, a sulfide- and sulfur-rich spring in southwestern Oklahoma; inner layers of microbial mats obtained from Sperm Pool, a high-temperature, low-pH pool (55°C, pH 2.5) in Yellowstone National Park; fresh bovine ruminal contents; and anaerobic freshwater pond sediments (Duck Pond) in Norman, Oklahoma. qPCR analysis indicated that SR1 members constitute a small fraction (<0.01%) of the microbial communities in Duck Pond and ruminal samples but constitute a significant fraction (11.6 and 48.7%) of the total number of bacterial 16S rRNA genes in Zodletone Spring and the inner layers of Sperm Pool microbial mat samples, respectively. By using SR1-specific fluorescent probes, filamentous cells were identified as the sole SR1 morphotype in all environments examined, with the exception of Sperm Pool, where a second bacillus morphotype was also identified. Using a full-cycle 16S rRNA approach, we show that each of these two morphotypes corresponds to a specific phylogenetic lineage identified in the Sperm Pool clone library. This work greatly expands the intralineage phylogenetic diversity within candidate division SR1 and provides valuable quantification and visualization tools that could be used for investigating the ecological roles, dynamics, and genomics of this as-yet-uncultured bacterial phylum.
机译:我们结合使用16S rRNA基因克隆文库调查,定量PCR(qPCR)分析和荧光原位杂交技术,研究了多个生境中SR1候选成员的多样性,丰度和分布。我们使用SR1特异性的16S rRNA基因引物,在四个不同的厌氧环境中鉴定了多个新的SR1谱系:俄克拉何马州西南部的Zodletone泉,富含硫化物和硫的泉水的沉积物;从黄石国家公园的高温,低pH池(55°C,pH 2.5)的Sperm Pool获得的微生物垫的内层;新鲜的牛瘤胃成分;和俄克拉荷马州诺曼的厌氧淡水池塘沉积物(鸭塘)。 qPCR分析表明,SR1成员在鸭塘和瘤胃样品中的微生物群落中占很小的比例(<0.01%),但在佐佐酮温泉(Zodletone)和精子池微生物垫样品的内层。通过使用SR1特异性荧光探针,在所有检查的环境中,丝状细胞被确定为唯一的SR1形态,除了精子池(Sperm Pool)之外,第二个芽孢杆菌也被鉴定为形态。使用全周期16S rRNA方法,我们显示这两个形态型分别对应于在精子库克隆文库中鉴定的特定系统发育谱系。这项工作大大扩展了候选分类SR1内的谱系内系统发育多样性,并提供了有价值的量化和可视化工具,可用于调查这种尚未培养的细菌门的生态作用,动力学和基因组学。

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