Qualitative and quantitative PCR methods using species-specific primer for detection and identification of wood rot fungi |
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Authors: | Sakae Horisawa Yoh Sakuma Shuichi Doi |
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Institution: | (1) Department of Environmental Systems Engineering, Faculty of Engineering, Kochi University of Technology, 185 Miyanokuchi, Kami, Kochi 782-8502, Japan;(2) Graduate School of Science and Engineering, Ehime University, Matsuyama 790-8577, Japan;(3) Graduate School of Life and Environmental Sciences, University of Tsukuba, Tsukuba, 305-8572 Ibaraki, Japan |
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Abstract: | Species-specific oligonucleotide primers for detecting wood rot fungi, Gloeophyllum trabeum, Trametes versicolor, Coniophora puteana, and Serpula lacrymans, and the primer detecting a group of related fungi to G. sepiarium were developed. These primer sequences were picked up from the internal transcribed spacer region between small-subunit rDNA
and large-subunit rDNA. The species selectivities of the developed primers were checked. Real-time polymerase chain reaction
(PCR) was carried out using these highly specific primers to quantitatively detect at least of 0.01 ng genome DNA of the target
species. This quantitative PCR was also used to differentiate the target species DNA from mixed species DNA. A PCR-based technique
using the species-specific primers would be applicable to multiple-sample assay in diagnosis of wood decay and to investigation
of environmental fungal populations.
Part of this article was presented at the International Symposium on Wood Science and Technology (IAWPC 2005), Yokohama, November
2005 |
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Keywords: | Species-specific primer Wood rot fungi Quantitative PCR Wood decay |
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