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不同有机改良剂施肥土壤中低有机碳培养基上细菌菌株的分离及部分特性研究。

Isolation and partial characterization of bacterial strains on low organic carbon medium from soils fertilized with different organic amendments.

机构信息

Plant Research International BV, Wageningen University and Research Center, Wageningen, The Netherlands.

出版信息

Microb Ecol. 2010 Nov;60(4):829-39. doi: 10.1007/s00248-010-9670-1. Epub 2010 Apr 27.

Abstract

A total of 720 bacterial strains were isolated from soils with four different organic amendment regimes on a low organic carbon (low-C) agar medium (10 µg C ml(-1)) traditionally used for isolation of oligotrophs. Organic amendments in combination with field history resulted in differences in dissolved organic carbon contents in these soils. There were negative correlations between total and dissolved organic carbon content and the number of isolates on low-C agar medium, whereas these correlations were absent for bacterial strains isolated from the same soil on high-C agar medium (1,000 µg C ml(-1)). Repeated transfers (up to ten times) of the isolates from low-C agar medium to fresh low- and high-C agar media were done to test for exclusive growth under oligotrophic conditions. The number of isolates exclusively growing under oligotrophic conditions dropped after each subsequent transfer from 241 after the first to 98 after the third transfer step. Identification on the basis of partial 16S rRNA gene sequences revealed that most of the 241 isolates (as well as the subset of 98 isolates) belong to widespread genera such as Streptomyces, Rhizobium, Bradyrhizobium, and Mesorhizobium, and the taxonomic composition of dominant genera changed from the first transfer step to the third. A selected subset of 17 isolates were further identified and characterized for exclusive growth on low-C agar medium. Two isolates continued to grow only on low-C agar medium up to the tenth transfer step and matched most closely with Rhizobium sullae and an uncultured bacterium on the basis of the almost full-length 16S rRNA gene. It was concluded that the vast majority of strains which are isolated on low-C agar media belong to the trophic group of microorganisms adapted to a "broad range" of carbon concentrations, including well-known and widespread bacterial genera. Oligotrophy is a physiological, not a taxonomic property, and can only be identified by cultural means so far. We showed that true oligotrophs that are unable to grow on high carbon media are rare and belong to genera that also contain broad-range and copiotrophic strains.

摘要

总共从四种不同有机改良剂处理的低有机碳(低-C)琼脂培养基(传统上用于分离寡营养菌的 10 µg C ml(-1))土壤中分离出 720 株细菌。有机改良剂与田间历史相结合导致这些土壤中溶解有机碳含量存在差异。低-C 琼脂培养基上总溶解有机碳含量与分离株数量之间呈负相关,而高-C 琼脂培养基(1,000 µg C ml(-1)) 上分离的细菌菌株则不存在这种相关性。从低-C 琼脂培养基中分离的菌株反复(最多 10 次)转移到新鲜的低-C 和高-C 琼脂培养基上,以测试在寡营养条件下的专性生长。从第一次转移到第三次转移后,专性在寡营养条件下生长的分离株数量从 241 株下降到 98 株。基于部分 16S rRNA 基因序列的鉴定表明,大多数 241 株(以及 98 株分离株的子集)属于广泛分布的属,如链霉菌属、根瘤菌属、慢生根瘤菌属和中慢生根瘤菌属,并且优势属的分类组成从第一转移步骤到第三转移步骤发生了变化。从 17 个分离株中选择了一个子集进一步鉴定并表征其在低-C 琼脂培养基上的专性生长。两个分离株在第十次转移步骤中仅在低-C 琼脂培养基上继续生长,并且基于几乎全长 16S rRNA 基因,与 Rhizobium sullae 和一种未培养细菌最为匹配。结论是,在低-C 琼脂培养基上分离出的绝大多数菌株属于适应“宽范围”碳浓度的微生物营养组,包括广为人知和广泛分布的细菌属。寡营养是一种生理特性,而不是一种分类特性,迄今为止只能通过培养手段来识别。我们表明,无法在高碳培养基上生长的真正寡营养菌很少,并且属于也含有广范围和富营养菌的属。

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