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A metagenome of a full-scale microbial community carrying out enhanced biological phosphorus removal

机译:全面微生物群落的元基因组可增强生物除磷能力

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摘要

Enhanced biological phosphorus removal (EBPR) is widely used for removal of phosphorus from wastewater. In this study, a metagenome (18.2 Gb) was generated using Illumina sequencing from a full-scale EBPR plant to study the community structure and genetic potential. Quantitative fluorescence in situ hybridization (qFISH) was applied as an independent method to evaluate the community structure. The results were in qualitative agreement, but a DNA extraction bias against gram positive bacteria using standard extraction protocols was identified, which would not have been identified without the use of qFISH. The genetic potential for community function showed enrichment of genes involved in phosphate metabolism and biofilm formation, reflecting the selective pressure of the EBPR process. Most contigs in the assembled metagenome had low similarity to genes from currently sequenced genomes, underlining the need for more reference genomes of key EBPR species. Only the genome of ‘Candidatus Accumulibacter', a genus of phosphorus-removing organisms, was closely enough related to the species present in the metagenome to allow for detailed investigations. Accumulibacter accounted for only 4.8% of all bacteria by qFISH, but the depth of sequencing enabled detailed insight into their microdiversity in the full-scale plant. Only 15% of the reads matching Accumulibacter had a high similarity (>95%) to the sequenced Accumulibacter clade IIA strain UW-1 genome, indicating the presence of some microdiversity. The differences in gene complement between the Accumulibacter clades were limited to genes for extracellular polymeric substances and phage-related genes, suggesting a selective pressure from phages on the Accumulibacter diversity.
机译:增强型生物除磷(EBPR)被广泛用于从废水中去除磷。在这项研究中,使用Illumina测序从完整的EBPR植物中生成了一个元基因组(18.2 Gb),以研究群落结构和遗传潜力。定量荧光原位杂交(qFISH)被用作评估群落结构的独立方法。结果在定性上一致,但是使用标准提取方案鉴定了针对革兰氏阳性细菌的DNA提取偏倚,如果不使用qFISH则无法确定。群落功能的遗传潜力表明参与磷酸盐代谢和生物膜形成的基因富集,反映了EBPR过程的选择性压力。组装后的基因组中的大多数重叠群与当前已测序基因组的基因具有较低的相似性,这表明需要更多关键EBPR物种的参考基因组。除磷生物的一个属“ Candidatus Accumulibacter”的基因组与该元基因组中存在的物种之间的关系非常密切,可以进行详细的研究。通过qFISH,积累细菌仅占所有细菌的4.8%,但是测序的深度使人们能够深入了解其在全规模植物中的微生物多样性。匹配Accumulibacter的读物中只有15%与测序的Accumulibacter clade IIA菌株UW-1基因组具有高度相似性(> 95%),表明存在一些微多样性。 Accumbacterbacter进化枝之间基因互补的差异仅限于细胞外聚合物质的基因和噬菌体相关的基因,表明噬菌体对Accumulibacter多样性具有选择性压力。

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