70 1,000,000, 難培養性微生物の実体とは i : lag time lag 1 6,7 3 Methanosaeta 2 Methanosaeta ii 1 lag time 6

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Journal of Environmental Biotechnology Vol. 7, No. 2, 69 73, 2007 総説 ( 特集 ) 難培養微生物とは何か? What Are Uncultured Microbes? YOICHI KAMAGATA 062 8517 2 17 2 1 TEL: 011-857-8914 FAX: 011-857-8915 E-mail: y.kamagata@aist.go.jp Research Institute of Genome-Based Biofactory, National Institute of Advanced Industrial Science and Technology (AIST), Toyohira, Sapporo, Hokkaido 062 8517, Japan キーワード Key words: uncultured microbes, cultivation, cell-cell communication, syntrophy 2007 10 26 2007 10 28 1. 難培養性微生物が認知されるまで 10 20 1990 20 10 16S rrna 2. 多くの微生物は難培養性微生物である 10 30 1 5,000 65 1990 16S rrna DNA 1 10 9 10,000 2 2005 10,000 1 1,000,000 3 1 1,000,000 3 1,000,000,000 0.0005

70 1,000,000,000 1880 4 1 0.1 5 3. 難培養性微生物の実体とは i : lag time 30 4 5 lag 1 6,7 3 Methanosaeta 2 Methanosaeta ii 1 lag time 6

71 8 iii 10 3 10 5 N- bacterial cytokine Resuscitation promoting factor Rpf iv oligotroph v Symbiobacterium thermophilum 9 Bacillus NH4 + 3 A Catellatibacterium nectariphililum B Sphingomonas 10,11 B A C D C G, F, I, J, K vi 3 in situ hybridization 12,13 2 Methanosaeta 3 A B

72 14 vii Buchnera 15 16 viii PCR 10 9 1 10 7 99 10 2 100,000 99 100,000 PCR 10 2 10 2 99 4. 再び難培養性微生物とは a b c 3 文 1) Whitman, W.B., D.C. Coleman, and W.J. Wiebe. 1998. Prokaryotes: the unseen majority. Proc. Natl. Acad. Sci. USA 95: 6578 6583. 2) Torsvik, V., J. Goksøyr, and F.L. Daae. 1990. High diversity in DNA of soil bacteria. Appl. Environ. Microbiol. 56: 782 787. 3) Gans, J., M. Wolinsky, and J. Dunbar. 2005. Computational improvements reveal great bacterial diversity and high metal toxicity in soil. Science 309: 1387 1389. 4) Bulloch, W. 2005 5) Amann, R.I., W. Ludwig, K.H. Schleifer. 1995. Phylogenetic identification and in situ detection of individual microbial cells without cultivation. Microbiol. Rev. 59: 143 169. 6) Qiu, Y-L., Y. Sekiguchi, H. Imachi, Y. Kamagata, I-C. Tseng, S-S. Cheng, A. Ohashi, and H. Harada. 2004. Identification and isolation of anaerobic, syntrophic p-phthalate isomersdegrading microbes from methanogenic sludges treating wastewater from terephthalate manufacturing. Appl. Environ. Microbiol. 70: 1617 1626. 7) Sakai, S., H. Imachi, Y. Sekiguchi, A. Ohashi, H. Harada, and Y. Kamagata. 2007. Isolation of key methanogens for global methane emission from rice paddy field: a novel isolate affiliated with a clone cluster, the Rice Cluster I. Appl. Environ. Microbiol. 73: 4326 4331. 8) Tamaki, H., Y. Sekiguchi, S. Hanada, K. Nakamura, N. Nomura, M. Matsumura, and Y. Kamagata. 2005. Comparative analysis of bacterial diversity in freshwater sediment of a shallow eutrophic lake as revealed by molecular and improved cultivation-based techniques. Appl. Environ. Microbiol. 71: 2162 2169. 9) Ohno, M., H. Shiratori, M.J. Park, Y. Saitoh, Y. Kumon, N. Yamashita, A. Hirata, H. Nishida, K. Ueda, and T. Beppu. 2000. Symbiobacterium thermophilum gen. nov., sp nov., a symbiotic thermophile that depends on co-culture with a Bacillus strain for growth. Int. J. Syst. Evol. Microbiol. 50: 1829 1832. 10) Tanaka, Y., S. Hanada, A. Manome, T. Tsuchida, R. Kurane, K. Nakamura, and Y. Kamagata. 2004. Catellatibacterium nectariphililum gen. nov., sp. nov., which requires a diffusible compound from a strain related to the genus Sphingomonas for vigorous growth. Int. J. Syst. Evol. Microbiol. 54: 955 959. 11) Tanaka, Y., S. Hanada, H. Tamaki, K. Nakamura, and Y. Kamagata. 2005. Isolation and identification of bacterial strains producing diffusible growth factor(s) for Catellibacterium nectariphilum strain AST4T. Microb. Environ. 20: 110 116. 12) Sekiguchi, Y., Y. Kamagata, K. Nakamura, A. Ohashi, and H. Harada. 1999. Fluorescence in situ hybridization using 16S rrna targeted oligonucleotides reveals localization of methanogens and selected uncultured bacteria in mesophilic and thermophilic sludge granules. Appl. Environ. Microbiol. 65: 1280 1288. 13) Imachi, H., Y. Sekiguchi, Y. Kamagata, A. Ohashi, and H. Harada. 2000. Cultivation and in situ detection of a thermo- 献

73 philic bacterium capable of oxidizing propionate in syntrophic association with hydrogenotrophic methanogens in a thermophilic methanogenic granular sludge. Appl. Environ. Microbiol. 66: 3608 3615. 14) Kamagata, Y., and H. Tamaki. 2005. Cultivation of uncultured fastidious microorganisms. Microb. Environ. 20: 85 91. 15) Shigenobu, S., H. Watanabe, M. Hattori, Y. Sakaki, and H. Ishikawa. 2000. Genome sequence of the endocellular bacterial symbiont of aphids Buchnera sp. APS. Nature 407: 81 86. 16) Shinzato, N., I. Watanabe, X-Y. Meng, Y. Sekiguchi, H. Tamaki, T. Matsui, and Y. Kamagata. 2007. Phylogenetic analysis and fluorescence in situ hybridization detection of archaeal and bacterial endosymbionts in the anaerobic ciliate Trimyema compressum. Microbial Ecol. (in press).