Rehydration conditions for isolation of high quality RNA from the lichen Lobaria pulmonaria

BMC Res Notes. 2014 Jul 10:7:442. doi: 10.1186/1756-0500-7-442.

Abstract

Background: The poikilohydric nature of lichens enables them to survive repeated episodes of desiccation by utilizing water when it becomes available. During rehydration, RNA-degrading endonucleases may be released, reducing RNA quantity and quality. Re-generation of a steady-state condition where RNA quantity and quality no longer fluctuate establishes a framework for development of new hypotheses for future investigations. Using Lobaria pulmonaria as a model species, the objective of this study was to compare the effect of different rehydration conditions on the quantity and quality of RNA from the rehydrated thallus.

Findings: Spectrophotometric measurements of total RNA and cDNA were performed for samples prepared from dry lichen or lichen after rehydration (0.5 h, 1 h, 2 h, 4 h or 24 h), with limited light and dark conditions, and at three temperatures (15°C, 20°C or 32°C) for some of these conditions. The results showed that rehydration of the thallus for 4 h at 20°C in light yielded the highest concentration and quality of RNA. A higher RNA concentration was obtained in light than in dark conditions, but the RNA quality was unaffected.

Conclusions: This study suggests that allowance of 4 h for thallus rehydration should be adequate to ensure complete recovery of transcription. After 4 h at 20°C further studies can be carried out on the RNA in this model species.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Desiccation
  • Lichens / chemistry*
  • Lichens / metabolism
  • Light
  • RNA Stability / radiation effects
  • RNA, Fungal / isolation & purification*
  • Specimen Handling
  • Temperature
  • Time Factors
  • Water / chemistry*

Substances

  • RNA, Fungal
  • Water