Identification of metabolically active proteobacterial and archaeal communities in the rumen by DNA- and RNA-derived 16S rRNA gene

J Appl Microbiol. 2013 Sep;115(3):644-53. doi: 10.1111/jam.12270. Epub 2013 Jul 1.

Abstract

Aims: To gain new insights into the metabolic contribution of bacterial group in the rumen.

Methods and results: Both DNA- and RNA-derived bacterial 16S ribosomal materials from bovine rumen contents were used as the template for bacterial community and analyse microbiota by three methods namely custom phylogenetic microarray, quantitative real-time PCR and denaturing gradient gel electrophoresis techniques. Bacterial analysis showed that genera affiliating with the Proteobacteria apparently made a greater metabolic contribution to rumen function than their population sizes indicated. Analysis of another rumen microbial group, the methanogens, using clone libraries for the expressed methyl coenzyme reductase subunit A (mcrA) revealed that an uncultivated methanogen clade contributes one-third of RNA-derived mcrA sequences based on a limited number of clones analysed. These uncultivated methanogen species were not observed in the mcrA gene library based on the DNA-derived sequences.

Conclusions: The comparison of results obtained from DNA- and RNA-derived materials suggests that some of the Proteobacteria and novel methanogen species appeared to be low in abundance in the rumen maintained on grain-based diets might play a greater role in rumen metabolism.

Significance and impact of the study: These studies provide the first report to compare high-throughput analysis of bacterial 16S rRNA genes from DNA- and RNA-derived materials to indicate differences that species make to community structure and metabolic activity.

Keywords: 16S rRNA gene; RNA; mcrA; metabolic activity; methanogen; rumen.

MeSH terms

  • Animals
  • Archaea / classification*
  • Archaea / genetics
  • Archaea / isolation & purification
  • Archaea / metabolism
  • Cattle
  • Denaturing Gradient Gel Electrophoresis
  • Genes, rRNA
  • Oligonucleotide Array Sequence Analysis
  • Oxidoreductases / genetics
  • Phylogeny
  • Proteobacteria / classification*
  • Proteobacteria / genetics
  • Proteobacteria / isolation & purification
  • Proteobacteria / metabolism
  • RNA, Ribosomal, 16S / genetics*
  • Real-Time Polymerase Chain Reaction
  • Rumen / metabolism
  • Rumen / microbiology*
  • Sequence Analysis, DNA
  • Sequence Analysis, RNA

Substances

  • RNA, Ribosomal, 16S
  • Oxidoreductases