Abstract

Community-level physiological profiling (CLPP) analyses from very diverse environments are frequently used with the aim of characterizing the metabolic versatility of whole environmental bacterial communities. While the limitations of the methodology for the characterization of whole communities are well known, we propose that CLPP combined with high-throughput sequencing and qPCR can be utilized to identify the copiotrophic, fast-growing fraction of the bacterial community of soil environments, where oligotrophic taxa are usually dominant. In the present work we have used this approach to analyze samples of litter and soil from a coniferous forest in the Czech Republic using BIOLOG GN2 plates. Monosaccharides and amino acids were utilized significantly faster than other C substrates, such as organic acids, in both litter and soil samples. Bacterial biodiversity in CLPP wells was significantly lower than in the original community, independently of the carbon source. Bacterial communities became highly enriched in taxa that typically showed low abundance in the original soil, belonging mostly to the Gammaproteobacteria and the genus Pseudomonas, indicating that the copiotrophic strains, favoured by the high nutrient content, are rare in forest litter and soil. In contrast, taxa abundant in the original samples were rarely found to grow at sufficient rates under the CLPP conditions. Our results show that CLPP is useful to detect copiotrophic bacteria from the soil environments and that bacterial growth is substrate specific.

Highlights

  • Microbial communities in forest soil have an essential role in organic matter decomposition and provide critical services to the ecosystem

  • The qPCR showed that there was no significant increase in the 16S copy numbers during plate incubation in the control wells with no substrate addition (“water“), while the 16S rRNA gene copy numbers in the DNA extracted from the plates were significantly higher in those wells where absorbance increased, indicating bacterial growth, than in the control wells (Fig 1)

  • Community-level physiological profiling (CLPP) using the BIOLOG system appears to be highly relevant for the screening of opportunistic r-strategists capable of fast growth that are competitive in mixed cultures, conditions to be expected at certain microniches, such as the tree root surfaces, which exude organic acids

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Summary

Introduction

Microbial communities in forest soil have an essential role in organic matter decomposition and provide critical services to the ecosystem. This role is of high importance considering the consequences of C storage or release and the fact that forest biomes currently represent a large C sink [1]. Utilization of various organic compounds by soil microbiota remains only partially understood or inferred from genome or metagenome sequencing [2]. Community-level physiological profiling (CLPP), based on sole C source utilization patterns, has been.

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