Effect of the Earthworms Lumbricus terrestris and Aporrectodea caliginosa on Bacterial Diversity in Soil

Abstract

Earthworms ingest large amounts of soil and have the potential to radically alter the biomass, activity, and structure of the soil microbial community. In this study, the diversity of eight bacterial groups from fresh soil, gut, and casts of the earthworms Lumbricus terrestris and Aporrectodea caliginosa were studied by single-strand conformation polymorphism (SSCP) analysis using both newly designed 16S rRNA gene-specific primer sets targeting Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, Deltaproteobacteria, Bacteroidetes, Verrucomicrobia, Planctomycetes, and Firmicutes and a conventional universal primer set for SSCP, with RNA and DNA as templates. In parallel, the study of the relative abundance of these taxonomic groups in the same samples was performed using fluorescence in situ hybridization. Bacteroidetes, Alphaproteobacteria, and Betaproteobacteria were predominant in communities from the soil and worm cast samples. Representatives of classes Flavobacteria and Sphingobacteria (Bacteroidetes) and Pseudomonas spp. (low-abundant Gammaproteobacteria) were detected in soil and worm cast samples with conventional and taxon-targeting SSCP and through the sequence analysis of 16S rRNA clone libraries. Physiologically active unclassified Sphingomonadaceae (Alphaproteobacteria) and Alcaligenes spp. (Betaproteobacteria) also maintained their diversities during transit through the earthworm intestine and were found on taxon-targeting SSCP profiles from the soil and worm cast samples. In conclusion, our results suggest that some specific bacterial taxonomic groups maintain their diversity and even increase their relative numbers during transit through the gastrointestinal tract of earthworms.

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Acknowledgments

This work was supported by the European Union Project “Biotic and Abiotic Mechanisms of TSE Infectivity Retention and Dissemination in Soil” TSE-SOIL-FATE (QLRT-2001-02493). K.N.T thanks the Fonds der Chemischen Industrie for generous support. B.A.B. and A.V.K. acknowledge the support of the Russian Foundation for Basic research (RFFI grant 06-04-48557). P.N.G. acknowledges the support from BiotechGenoMik project of the Federal Ministry for Science and Education (BMBF).

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Correspondence to Taras Y. Nechitaylo.

Electronic supplementary material

Supplementary information contains phylogenetic analysis of bacterial 16S rRNA clones from the libraries, TT-SSCP profiles of a number of bacterial groups and phylogenetic analysis of ribotypes from the SSCP profiles, and details on statistical analysis of FISH counts.

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Supplementary SSCP Analysis

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Nechitaylo, T.Y., Yakimov, M.M., Godinho, M. et al. Effect of the Earthworms Lumbricus terrestris and Aporrectodea caliginosa on Bacterial Diversity in Soil. Microb Ecol 59, 574–587 (2010). https://doi.org/10.1007/s00248-009-9604-y

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Keywords

  • Firmicutes
  • Bacteroidetes
  • Gammaproteobacteria
  • Alphaproteobacteria
  • Betaproteobacteria