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

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

doi:10.1264/jsme2.23.248

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

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

Department of Microbiology and Immunology

Research output: Contribution to journal › Article › peer-review

16 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 language	English
Pages (from-to)	248-252
Number of pages	5
Journal	Microbes and Environments
Volume	23
Issue number	3
DOIs	https://doi.org/10.1264/jsme2.23.248
Publication status	Published - 2008

Keywords

DGGE
DNA isolation
Epiphytic bacteria
Phyllosphere

ASJC Scopus subject areas

Ecology, Evolution, Behavior and Systematics
Soil Science
Plant Science

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10.1264/jsme2.23.248

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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.",

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AU - Suda, Wataru

AU - Oto, Michiei

AU - Amachi, Seigo

AU - Shinoyama, Hirofumi

AU - Shishido, Masahiro

PY - 2008

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AB - 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.

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KW - Epiphytic bacteria

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

Abstract

Keywords

ASJC Scopus subject areas

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