Abstract
To appreciate the functional diversity of communities of soil eukaryotic micro-organisms we evaluated an experimental approach based on the construction and screening of a cDNA library using polyadenylated mRNA extracted from a forest soil. Such a library contains genes that are expressed by each of the different organisms forming the community and represents its metatranscriptome. The diversity of the organisms that contributed to this library was evaluated by sequencing a portion of the 18S rDNA gene amplified from either soil DNA or reverse-transcribed RNA. More than 70% of the sequences were from fungi and unicellular eukaryotes (protists) while the other most represented group was the metazoa. Calculation of richness estimators suggested that more than 180 species could be present in the soil samples studied. Sequencing of 119 cDNA identified genes with no homologues in databases (32%) and genes coding proteins involved in different biochemical and cellular processes. Surprisingly, the taxonomic distribution of the cDNA and of the 18S rDNA genes did not coincide, with a marked under-representation of the protists among the cDNA. Specific genes from such an environmental cDNA library could be isolated by expression in a heterologous microbial host, Saccharomyces cerevisiae. This is illustrated by the functional complementation of a histidine auxotrophic yeast mutant by two cDNA originating possibly from an ascomycete and a basidiomycete fungal species. Study of the metatranscriptome has the potential to uncover adaptations of whole microbial communities to local environmental conditions. It also gives access to an abundant source of genes of biotechnological interest.
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Acknowledgements
This work was supported by grants from the University Lyon 1 (BQR 2005), the French Ministry of Ecology and Sustainable Development (PNETOX programme) and the IFR 41. Sequencing of ribosomal genes was supported by an ‘Etude et Appuis’ grant of the Bureau des Ressources Génétiques. We thank Jacques Guinberteau and the other members of the INRA of Bordeaux for their help in field work and the determination of fungal species and the DTAMB of the University Lyon 1 for the use of the Agilent Bioanalyzer.
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Bailly, J., Fraissinet-Tachet, L., Verner, MC. et al. Soil eukaryotic functional diversity, a metatranscriptomic approach. ISME J 1, 632–642 (2007). https://doi.org/10.1038/ismej.2007.68
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DOI: https://doi.org/10.1038/ismej.2007.68
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