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Applied and Environmental Microbiology, May 2000, p. 2199-2207, Vol. 66, No. 5
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Localization of Symbiotic Clostridia in the Mixed Segment of the Termite Nasutitermes takasagoensis (Shiraki)

Gaku Tokuda,1 Ikuo Yamaoka,2 and Hiroaki Noda3,*

Bio-oriented Technology Research Advancement Institution, Nisshin-cyo, Omiya, Saitama 331-8537,1 Department of Biology, Physics and Informatics, Faculty of Science, Yamaguchi University, Yoshida, Yamaguchi 753-8512,2 and National Institute of Sericultural and Entomological Science, Owashi, Tsukuba, Ibaraki 305-8634,3 Japan

Received 31 August 1999/Accepted 7 February 2000

Phylogeny and the distribution of symbiotic bacteria in the mixed segment of the wood-eating termite Nasutitermes takasagoensis (Shiraki) were studied. Bacterial 16S rRNA genes (rDNA) were amplified from the mixed segment of the gut by PCR, and two kinds of sequences were identified. The phylogenetic tree was constructed by neighbor-joining and maximum parsimony methods to identify symbionts harbored in the mixed segment. They are classified as low-G+C-content gram-positive bacteria and are most closely related to the genus Clostridium. The distribution of these bacteria throughout the whole gut was examined by PCR using specific primers, which suggested that they are confined to the mixed segment despite the presence of bacteria throughout the gut. In situ hybridization indicated that the symbiotic bacteria were localized to the ectoperitrophic space between the midgut wall and the peritrophic membrane in the mixed segment. Electron microscopy revealed the close association between these bacteria and the mesenteric epithelium, suggesting that they have some interactions with the gut tissue of termites.

* Corresponding author. Mailing address: National Institute of Sericultural and Entomological Science, Owashi, Tsukuba, Ibaraki 305-8634, Japan. Phone: (81) 298-38-6109. Fax: (81) 298-38-6028. E-mail: hnada{at}nises.affrc.go.jp.

Applied and Environmental Microbiology, May 2000, p. 2199-2207, Vol. 66, No. 5
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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