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Specific Detection and Real-Time PCR Quantification of Potentially Mycophagous Bacteria Belonging to the Genus Collimonas in Different Soil Ecosystems

机译:不同土壤生态系统中属于准直属属的潜在嗜食菌细菌的特异性检测和实时PCR定量

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摘要

The bacterial genus Collimonas has the remarkable characteristic that it grows at the expense of living fungal hyphae under laboratory conditions. Here, we report the first field inventory of the occurrence and abundance of Collimonas in soils (n = 45) with naturally different fungal densities, which was performed in order to test the null hypothesis that there is a relationship between the presence of Collimonas and fungal biomass. Estimates of fungal densities were based on ergosterol measurements. Each soil was also characterized in terms of its physical and chemical properties and vegetation and management types. Culturable Collimonas was identified in plate-spread soil samples by its ability to clear colloidal chitin, in combination with a Collimonas-specific restriction fragment length polymorphism analysis of 16S rRNA PCR amplified from individual colonies. Using this approach, we found culturable collimonads only in (semi)natural grasslands. A real-time PCR assay for the specific quantification of Collimonas 16S rRNA in total soil DNA was developed. Collimonas was detectable in 80% of the soil samples, with densities up to 105 cells g−1 (dry weight) soil. The numbers of Collimonas cells per gram of soil were consistently lowest in fungus-poor arable soils and, surprisingly, also in fungus-rich organic layers of forest soils. When all soils were included, no significant correlation was observed between the number of Collimonas cells and ergosterol-based soil fungal biomass. Based on this result, we rejected our null hypothesis, and possible explanations for this were addressed.
机译:细菌科利莫纳氏菌具有显着的特征,即它在实验室条件下以牺牲活菌丝的形式生长。在这里,我们报告了自然界不同​​真菌密度的土壤(n = 45)中Collimonas的发生和丰度的第一个现场调查,其目的是检验零假设:Collimonas的存在与真菌之间存在关系生物质。真菌密度的估计基于麦角固醇测量。还根据每种土壤的物理和化学性质以及植被和管理类型对其进行了表征。通过清除散布的甲壳质的能力,结合从单个菌落扩增的16S rRNA PCR的Collimonas特异性限制性片段长度多态性分析,在板状土壤样品中鉴定出可培养的Collimonas。使用这种方法,我们仅在(半)天然草原上发现了可培养的准直虫。建立了实时定量PCR技术,用于对土壤中总DNA中的Collimonas 16S rRNA进行特异性定量。在80%的土壤样品中可检测到Collimonas,密度高达10 5 个细胞g -1 (干重)土壤。真菌贫乏的土壤以及森林土壤富含真菌的有机层中,每克土壤中的Collimonas细胞数量始终是最低的。当包括所有土壤时,Collimonas细胞的数量与基于麦角固醇的土壤真菌生物量之间没有显着相关性。基于此结果,我们拒绝了原假设,并提出了对此的可能解释。

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