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Appl Environ Microbiol, May 1998, p. 1680-1687, Vol. 64, No. 5
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Phylogenetic Evidence for the Existence of Novel Thermophilic Bacteria in Hot Spring Sulfur-Turf Microbial Mats in Japan

Hiroyuki Yamamoto,1,* Akira Hiraishi,2 Kenji Kato,3 Hiroshi X. Chiura,4 Yonosuke Maki,5 and Akira Shimizu6

Department of Microbiology, St. Marianna University School of Medicine, Kawasaki 216,1 Department of Ecological Engineering, Toyohashi University of Technology, Toyohashi 441,2 Laboratory of Biology, School of Allied Medical Sciences, Shinshu University, Matsumoto 390,3 Department of Biology, Division of Natural Sciences, International Christian University, Mitaka 181,4 Laboratory of Biology, Faculty of Humanities and Social Sciences, Iwate University, Morioka 020,5 and Department of Biology, Faculty of Science, Nara Women's University, Nara 630,6 Japan

Received 6 October 1997/Accepted 2 March 1998

So-called sulfur-turf microbial mats, which are macroscopic white filaments or bundles consisting of large sausage-shaped bacteria and elemental sulfur particles, occur in sulfide-containing hot springs in Japan. However, no thermophiles from sulfur-turf mats have yet been isolated as cultivable strains. This study was undertaken to determine the phylogenetic positions of the sausage-shaped bacteria in sulfur-turf mats by direct cloning and sequencing of 16S rRNA genes amplified from the bulk DNAs of the mats. Common clones with 16S rDNA sequences with similarity levels of 94.8 to 99% were isolated from sulfur-turf mat samples from two geographically remote hot springs. Phylogenetic analysis showed that the phylotypes of the common clones formed a major cluster with members of the Aquifex-Hydrogenobacter complex, which represents the most deeply branching lineage of the domain bacteria. Furthermore, the bacteria of the sulfur-turf mat phylotypes formed a clade distinguishable from that of other members of the Aquifex-Hydrogenobacter complex at the order or subclass level. In situ hybridization with clone-specific probes for 16S rRNA revealed that the common phylotype of sulfur-turf mat bacteria is that of the predominant sausage-shaped bacteria.

* Corresponding author. Mailing address: Department of Microbiology, St. Marianna University School of Medicine, Sugao 2-16-1 Miyamae, Kawasaki, Kanagawa 216-8511. Japan. Phone: 81-44-977-8111. Fax: 81-44-977-7818. E-mail: kyama{at}marianna-u.ac.jp.

Appl Environ Microbiol, May 1998, p. 1680-1687, Vol. 64, No. 5
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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