Isolation of fungal DNA from plant tissues and removal of DNA amplification inhibitors

Dean K. Malvick, Eric Grunden

Research output: Contribution to journalArticlepeer-review

31 Scopus citations

Abstract

Specific primers and the polymerase chain reaction (PCR) are increasingly used for detection of fungi in plants. Detection depends on isolation of DNA that is free of compounds that inhibit amplification. We report on the significance of inhibition in soybean stems with a focus on studies of the vascular fungal pathogen Phialophora gregata. A simple DNA extraction procedure based on a FastDNA® kit is described that allows consistent detection of fungi in soybean stems using PCR. The addition of polyvinylpyrrolidone and supplemental DNA purification steps overcame inhibition in over 90% of samples. These methods should also facilitate studies with other plant and fungal species.

Original languageEnglish (US)
Pages (from-to)958-960
Number of pages3
JournalMolecular Ecology Notes
Volume5
Issue number4
DOIs
StatePublished - Dec 1 2005

Keywords

  • DNA
  • Diagnosis
  • Fungi
  • Inhibition
  • PCR
  • Soybean

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