Analysis of fungal community in decayed wood by PCR-based denaturing gradient gel electrophoresis

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  • NAKADA Yuji
    Faculty of Agriculture, Tokyo University of Agriculture and Technology
  • KUSUMI Asako
    Faculty of Agriculture, Tokyo University of Agriculture and Technology
  • NAKABA Satoshi
    Faculty of Agriculture, Tokyo University of Agriculture and Technology
  • KATAYAMA Yoko
    Faculty of Agriculture, Tokyo University of Agriculture and Technology
  • FUNADA Ryo
    Faculty of Agriculture, Tokyo University of Agriculture and Technology
  • FUKUDA Kiyoharu
    Faculty of Agriculture, Tokyo University of Agriculture and Technology
  • YOSHIDA Makoto
    Faculty of Agriculture, Tokyo University of Agriculture and Technology

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Other Title
  • 変性剤濃度勾配ゲル電気泳動法を利用した腐朽木材中に存在する真菌類の菌叢解析
  • ヘンセイザイ ノウド コウバイ ゲル デンキ エイドウホウ オ リヨウ シタ フキュウ モクザイ チュウ ニ ソンザイ スル シン キンルイ ノ キンソウカイセキ

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Abstract

To understand the mechanism of wood decay, fungal community in decayed wood was analyzed by denaturing gradient gel electrophoresis(DGGE). The genomic DNAs extracted from fifteen species of filamentous fungi were used as a template for PCR to amplify internal transcribed spacer(ITS)in ribosomal DNA. The amplification products were analyzed by electrophoresis on agarose gel, and the single band was found around 400 bp in all samples. On the other hand, different migration rates of the bands were clearly observed in DGGE analysis, indicating that DNA fragments of ITS regions from different fungi can be distinguished on the gel. In the next experiment, genomic DNAs were prepared from six of decayed woody materials, and ITS regions of wood inhabiting fungi were amplified by PCR following random amplification of genomic DNA by using Phi29 DNA polymerase. More bands were shown in DGGE analysis compared with agarose gel electrophoresis. Sequence analysis of PCR products suggested the existence of various fungi in these samples. These results indicate that the method is useful for the analysis of fungal community existed in the decayed wood.

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