Comparison of 16S rRNA and 16S rDNA T-RFLP approaches to study bacterial communities in soil microcosms treated with chromate as perturbing agent

Microb Ecol. 2005 Oct;50(3):375-84. doi: 10.1007/s00248-004-0222-4. Epub 2005 Nov 3.

Abstract

Transcripts of ribosomal RNA have been used for assessing the structure and dynamics of active bacterial populations; however, it remains unclear whether the information provided by community profiling derived from RNA is different from that derived from DNA, particularly when a selective pressure is applied on the bacterial community. In the present work, terminal-restriction fragment length polymorphism (T-RFLP) community profiles based on DNA and RNA extracted from soil microcosms treated with a toxic concentration of chromate were compared. Microcosms of a nonpolluted agricultural soil and of a heavy-metal-rich soil (serpentine) were treated with chromate and DNA and RNA were extracted. T-RFLP analysis was performed on amplified and retro-amplified 16SrRNA gene sequences, and band profiles obtained from samples of DNA and of RNA were compared. Some of the T-RFLP bands, identified as peculiar peaks in the profiles, were cloned and sequenced for taxonomic interpretation. Results indicated that: (1) community profiles derived from RNA and DNA were partly overlapping; (2) there was a strong correlation between the dynamics shown by RNA- and DNA-based T-RFLP profiles; (3) chromate addition exerted a clear effect on both agricultural and serpentine soil bacterial communities, either at the DNA and at the RNA level; however, the profiles derived from RNA showed sharper differences between treated and control samples than that of DNA-based profiles.

MeSH terms

  • Agriculture
  • Asbestos, Serpentine
  • Bacteria / classification
  • Bacteria / genetics
  • Bacteria / isolation & purification*
  • Chromates
  • DNA, Bacterial / genetics
  • Polymorphism, Restriction Fragment Length*
  • RNA, Bacterial / genetics
  • RNA, Ribosomal, 16S / genetics
  • Sensitivity and Specificity
  • Soil / analysis
  • Soil Microbiology*
  • Soil Pollutants
  • Species Specificity

Substances

  • Asbestos, Serpentine
  • Chromates
  • DNA, Bacterial
  • RNA, Bacterial
  • RNA, Ribosomal, 16S
  • Soil
  • Soil Pollutants