A direct method to isolate dna from phyllosphere microbial communities without disrupting leaf tissues

Wataru Suda, Michiei Oto, Seigo Amachi, Hirofumi Shinoyama, Masahiro Shishido

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

We developed a method for direct DNA isolation from phyllosphere microbial communities, designated Direct-DIP. This method comprises DNA extraction from non-shredded leaves with benzyl chloride, and DNA purification by gel filtration. Scanning electron microscopy showed that epiphytic microorganisms were completely removed from the leaf surface after benzyl chloride treatment, while microstructures of the leaf were not damaged. Clear DGGE profiles were obtained regardless of the plant species. Shannon diversity indices of DGGE profiles by Direct-DIP were higher than those by a conventional method. Our findings suggest that Direct-DIP is a rapid, simple, and cost-effective method of extracting DNA from phyllosphere microbial communities.

Original languageEnglish
Pages (from-to)248-252
Number of pages5
JournalMicrobes and Environments
Volume23
Issue number3
DOIs
Publication statusPublished - 2008
Externally publishedYes

Fingerprint

phyllosphere
microbial communities
microbial community
DNA
leaves
chlorides
chloride
methodology
diversity index
microstructure
purification
scanning electron microscopy
gel
microorganism
gels
microorganisms
tissue
method
tissues
cost

Keywords

  • DGGE
  • DNA isolation
  • Epiphytic bacteria
  • Phyllosphere

ASJC Scopus subject areas

  • Ecology, Evolution, Behavior and Systematics
  • Soil Science
  • Plant Science

Cite this

A direct method to isolate dna from phyllosphere microbial communities without disrupting leaf tissues. / Suda, Wataru; Oto, Michiei; Amachi, Seigo; Shinoyama, Hirofumi; Shishido, Masahiro.

In: Microbes and Environments, Vol. 23, No. 3, 2008, p. 248-252.

Research output: Contribution to journalArticle

Suda, Wataru ; Oto, Michiei ; Amachi, Seigo ; Shinoyama, Hirofumi ; Shishido, Masahiro. / A direct method to isolate dna from phyllosphere microbial communities without disrupting leaf tissues. In: Microbes and Environments. 2008 ; Vol. 23, No. 3. pp. 248-252.
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