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与β-亚群氨氧化细菌相关的土壤和海洋16S rRNA基因序列的分子多样性

Molecular diversity of soil and marine 16S rRNA gene sequences related to beta-subgroup ammonia-oxidizing bacteria.

作者信息

Stephen J R, McCaig A E, Smith Z, Prosser J I, Embley T M

机构信息

Department of Zoology, Natural History Museum, London, United Kingdom.

出版信息

Appl Environ Microbiol. 1996 Nov;62(11):4147-54. doi: 10.1128/aem.62.11.4147-4154.1996.

Abstract

We have conducted a preliminary phylogenetic survey of ammonia-oxidizing beta-proteobacteria, using 16S rRNA gene libraries prepared by selective PCR and DNA from acid and neutral soils and polluted and nonpolluted marine sediments. Enrichment cultures were established from samples and analyzed by PCR. Analysis of 111 partial sequences of c. 300 bases revealed that the environmental sequences formed seven clusters, four of which are novel, within the phylogenetic radiation defined by cultured autotrophic ammonia oxidizers. Longer sequences from 13 cluster representatives support their phylogenetic positions relative to cultured taxa. These data suggest that known taxa may not be representative of the ammonia-oxidizing beta-proteobacteria in our samples. Our data provide further evidence that molecular and culture-based enrichment methods can select for different community members. Most enrichments contained novel Nitrosomonas-like sequences whereas novel Nitrosospira-like sequences were more common from gene libraries of soils and marine sediments. This is the first evidence for the occurrence of Nitrosospira-like strains in marine samples. Clear differences between the sequences of soil and marine sediment libraries were detected. Comparison of 16S rRNA sequences from polluted and nonpolluted sediments provided no strong evidence that the community composition was determined by the degree of pollution. Soil clone sequences fell into four clusters, each containing sequences from acid and neutral soils in varying proportions. Our data suggest that some related strains may be present in both samples, but further work is needed to resolve whether there is selection due to pH for particular sequence types.

摘要

我们利用通过选择性PCR制备的16S rRNA基因文库以及来自酸性和中性土壤、污染和未污染海洋沉积物的DNA,对氨氧化β-变形菌进行了初步的系统发育调查。从样品中建立富集培养物并通过PCR进行分析。对约300个碱基的111个部分序列进行分析后发现,环境序列在由培养的自养氨氧化菌定义的系统发育辐射范围内形成了七个簇,其中四个是新的。来自13个簇代表的更长序列支持了它们相对于培养分类群的系统发育位置。这些数据表明,已知的分类群可能不能代表我们样品中的氨氧化β-变形菌。我们的数据进一步证明,基于分子和培养的富集方法可以选择不同的群落成员。大多数富集培养物包含新的类硝化单胞菌序列,而新的类硝化螺菌序列在土壤和海洋沉积物的基因文库中更为常见。这是海洋样品中存在类硝化螺菌菌株的首个证据。检测到土壤和海洋沉积物文库序列之间存在明显差异。对污染和未污染沉积物的16S rRNA序列进行比较,没有提供有力证据表明群落组成由污染程度决定。土壤克隆序列分为四个簇,每个簇包含来自酸性和中性土壤的不同比例的序列。我们的数据表明,两个样品中可能都存在一些相关菌株,但需要进一步研究来确定是否存在因pH值导致的特定序列类型的选择。

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