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Applied and Environmental Microbiology, December 2005, p. 8811-8817, Vol. 71, No. 12
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.12.8811-8817.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Endosymbiotic Bacteroidales Bacteria of the Flagellated Protist Pseudotrichonympha grassii in the Gut of the Termite Coptotermes formosanus

Satoko Noda,1 Toshiya Iida,2 Osamu Kitade,3 Hideaki Nakajima,2,4 Toshiaki Kudo,2,5 and Moriya Ohkuma1,2*

PRESTO, Japan Science and Technology Agency (JST), Wako, Saitama 351-0198, Japan,1 Environmental Molecular Biology Lab, RIKEN, Wako, Saitama 351-0198, Japan,2 Natural History Lab, Faculty of Science, Ibaraki University, Mito, Ibaraki 310-8512, Japan,3 Department of Applied Chemistry, Toyo University, Kawagoe, Saitama 350-8585, Japan,4 Graduate School of Integrated Science, Yokohama City University, Suehiro, Tsurumi-ku, Yokohama 230-0045, Japan5

Received 1 July 2005/ Accepted 25 August 2005

A unique lineage of bacteria belonging to the order Bacteroidales was identified as an intracellular endosymbiont of the protist Pseudotrichonympha grassii (Parabasalia, Hypermastigea) in the gut of the termite Coptotermes formosanus. We identified the 16S rRNA, gyrB, elongation factor Tu, and groEL gene sequences in the endosymbiont and detected a very low level of sequence divergence (<0.9% of the nucleotides) in the endosymbiont population within and among protist cells. The Bacteroidales endosymbiont sequence was affiliated with a cluster comprising only sequences from termite gut bacteria and was not closely related to sequences identified for members of the Bacteroidales attached to the cell surfaces of other gut protists. Transmission electron microscopy showed that there were numerous rod-shaped bacteria in the cytoplasm of the host protist, and we detected the endosymbiont by fluorescence in situ hybridization (FISH) with an oligonucleotide probe specific for the 16S rRNA gene identified. Quantification of the abundance of the Bacteroidales endosymbiont by sequence-specific cleavage of rRNA with RNase H and FISH cell counting revealed, surprisingly, that the endosymbiont accounted for 82% of the total bacterial rRNA and 71% of the total bacterial cells in the gut community. The genetically nearly homogeneous endosymbionts of Pseudotrichonympha were very abundant in the gut symbiotic community of the termite.

* Corresponding author. Mailing address: Environmental Molecular Biology Laboratory, RIKEN, Hirosawa 2-1, Wako, Saitama 351-0198, Japan. Phone: 81-48-467-9648. Fax: 81-48-462-4672. E-mail: mohkuma{at}riken.jp.

Applied and Environmental Microbiology, December 2005, p. 8811-8817, Vol. 71, No. 12
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.12.8811-8817.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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