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ID 53977
フルテキストURL
122882.zip 2.08 MB
著者
Tani, Akio Institute of Plant Science and Resources, Okayama University
Nurettin, Sahin Faculty of Education, Mugla Sitki Kocman University
Matsuyama, Yumiko
Enomoto, Takashi Institute of Plant Science and Resources, Okayama University
Nishimura, Naoki Botanical Garden, Okayama University of Science
Yokota, Akira Faculty of Mathematics and Natural Sciences, University of Indonesia
Kimbara, Kazuhide Faculty of Engineering, Shizuoka University
抄録
Methylobacterium species are ubiquitous α-proteobacteria that reside in the phyllosphere and are fed by methanol that is emitted from plants. In this study, we applied whole-cell matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analysis (WC-MS) to evaluate the diversity of Methylobacterium species collected from a variety of plants. The WC-MS spectrum was reproducible through two weeks of cultivation on different media. WC-MS spectrum peaks of M. extorquens strain AM1 cells were attributed to ribosomal proteins, but those were not were also found. We developed a simple method for rapid identification based on spectra similarity. Using all available type strains of Methylobacterium species, the method provided a certain threshold similarity value for species-level discrimination, although the genus contains some type strains that could not be easily discriminated solely by 16S rRNA gene sequence similarity. Next, we evaluated the WC-MS data of approximately 200 methylotrophs isolated from various plants with MALDI Biotyper software (Bruker Daltonics). Isolates representing each cluster were further identified by 16S rRNA gene sequencing. In most cases, the identification by WC-MS matched that by sequencing, and isolates with unique spectra represented possible novel species. The strains belonging to M. extorquens, M. adhaesivum, M. marchantiae, M. komagatae, M. brachiatum, M. radiotolerans, and novel lineages close to M. adhaesivum, many of which were isolated from bryophytes, were found to be the most frequent phyllospheric colonizers. The WC-MS technique provides emerging high-throughputness in the identification of known/novel species of bacteria, enabling the selection of novel species in a library and identification without 16S rRNA gene sequencing.
備考
This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
発行日
2012-07-12
出版物タイトル
PLOS ONE
7巻
7号
出版者
PUBLIC LIBRARY SCIENCE
開始ページ
e40784
ISSN
1932-6203
資料タイプ
学術雑誌論文
オフィシャル URL
http://dx.doi.org/10.1371/journal.pone.0040784
言語
English
著作権者
© 2012 Tani et al.
論文のバージョン
publisher
査読
有り
DOI
PubMed ID
Web of Sience KeyUT