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Molecular Analysis of Bacterial Community Structure and Diversity in Unimproved and Improved Upland Grass Pastures

机译:未经改良和改良的旱地草场细菌群落结构和多样性的分子分析

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摘要

Bacterial community structure and diversity in rhizospheres in two types of grassland, distinguished by both plant species and fertilization regimen, were assessed by performing a 16S ribosomal DNA (rDNA) sequence analysis of DNAs extracted from triplicate soil plots. PCR products were cloned, and 45 to 48 clones from each of the six libraries were partially sequenced. Phylogenetic analysis of the resultant 275 clone sequences indicated that there was considerable variation in abundance in replicate unfertilized, unimproved soil samples and fertilized, improved soil samples but that there were no significant differences in the abundance of any phylogenetic group. Several clone sequences were identical in the 16S rDNA region analyzed, and the clones comprised eight pairs of duplicate clones and two sets of triplicate clones. Many clones were found to be most closely related to environmental clones obtained in other studies, although three clones were found to be identical to culturable species in databases. The clones were clustered into operational taxonomic units at a level of sequence similarity of >97% in order to quantify diversity. In all, 34 clusters containing two or more sequences were identified, and the largest group contained nine clones. A number of diversity, dominance, and evenness indices were calculated, and they all indicated that diversity was high, reflecting the low coverage of rDNA libraries achieved. Differences in diversity between sample types were not observed. Collector’s curves, however, indicated that there were differences in the underlying community structures; in particular, there was reduced diversity of organisms of the α subdivision of the class Proteobacteria (α-proteobacteria) in improved soils.
机译:通过对从一式三份的土壤样地中提取的DNA进行16S核糖体DNA(rDNA)序列分析,评估了以植物种类和施肥方案区分的两种类型草地的细菌群落结构和根际多样性。克隆了PCR产物,并对来自六个文库中每个文库的45至48个克隆进行了部分测序。对得到的275个克隆序列的系统进化分析表明,在未施肥,未经改良的土壤样品和经改良和改良的土壤样品中,丰度存在很大差异,但任何系统发育组的丰度均无显着差异。在分析的16S rDNA区域中,几个克隆序列是相同的,并且该克隆包含八对重复的克隆和两组一式三份的克隆。尽管发现三个克隆与数据库中的可培养物种相同,但发现许多克隆与其他研究中获得的环境克隆关系最密切。将克隆按> 97%的序列相似性水平聚类为可操作的分类单位,以量化多样性。总共鉴定出包含两个或多个序列的34个簇,最大的一组包含9个克隆。计算了许多多样性,优势度和均匀性指数,它们都表明多样性很高,这反映出所实现的rDNA文库覆盖率较低。没有观察到样品类型之间多样性的差异。然而,收藏家的曲线表明,底层社区的结构存在差异。特别是在改良的土壤中,Proteobacteria类(α-proteobacteria)的α细分有机体的多样性降低了。

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