Phylogenetic analysis of Metabacterium polyspora: clues to the evolutionary origin of daughter cell production in Epulopiscium species, the largest bacteria.

J Bacteriol. 1996 March; 178(5): 1451–1456.

PMCID: PMC177821

E R Angert, A E Brooks, and N R Pace

Department of Biology, Indiana University, Bloomington 47405, USA.

This article has been cited by other articles in PMC.

Abstract

It is rare that there are molecular clues to the evolutionary origin of developmental traits. We have encountered an evolutionary juxtaposition that may explain the origin of the unique replicative morphology of Epulopiscium spp., the largest known bacteria, which reproduce by the internal production of multiple live offspring. We report here a 16S rRNA-based phylogenetic analysis of Metabacterium polyspora, a multiple-endospore-forming, uncultivated inhabitant of guinea pig cecum. Cells of M. polyspora were harvested from cecum contents by sedimentation in a Ficoll gradient and lysed. The bacterial 16S rRNA genes of this lysate were amplified by PCR. Sequence analysis of the cloned PCR products revealed two dominant, closely related 16S rRNA types. In situ hybridization of cecum contents with fluorescently labeled oligonucleotides, diagnostic of these two sequences, showed that they represent distinct strains of M. polyspora. Phylogenetic analyses of the sequences showed that M. polyspora is closely related to Epulopiscium spp. On the basis of this result and other correlations, we propose that the process of sporulation was modified in a predecessor of Epulopiscium spp. to produce live offspring instead of quiescent endospores.

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Angert, ER; Clements, KD; Pace, NR. The largest bacterium. Nature. 1993 Mar 18;362(6417):239–241. [PubMed]
Brosius, J; Palmer, ML; Kennedy, PJ; Noller, HF. Complete nucleotide sequence of a 16S ribosomal RNA gene from Escherichia coli. Proc Natl Acad Sci U S A. 1978 Oct;75(10):4801–4805. [PubMed]
Chapman, GB; Slob-van Herk, A; Eguía, JM. The occurrence of disporous Bacillus thuringiensis cells. Antonie Van Leeuwenhoek. 1992 May;61(4):265–268. [PubMed]
Chase, DG; Erlandsen, SL. Evidence for a complex life cycle and endospore formation in the attached, filamentous, segmented bacterium from murine ileum. J Bacteriol. 1976 Jul;127(1):572–583. [PubMed]
Collins, MD; Lawson, PA; Willems, A; Cordoba, JJ; Fernandez-Garayzabal, J; Garcia, P; Cai, J; Hippe, H; Farrow, JA. The phylogeny of the genus Clostridium: proposal of five new genera and eleven new species combinations. Int J Syst Bacteriol. 1994 Oct;44(4):812–826. [PubMed]
DELAPORTE, B. ETUDE COMPAR'EE DE GRANDS SPIRILLES FORMANT DES SPORES: SPOROSPIRILLUM (SPIRILLUM) PRAECLARUM (COLLIN) N.G., SPOROSPIRILLUM GYRINI N.SP. ET SPOROSPIRILLUM BISPORUM N.SP. Ann Inst Pasteur (Paris). 1964 Aug;107:246–262. [PubMed]
DELAPORTE, B. ETUDE DESCRIPTIVE DE BACT'ERIES DE TR'ES GRANDES DIMENSIONS. Ann Inst Pasteur (Paris). 1964 Dec;107:845–862. [PubMed]
DeLong, EF; Wickham, GS; Pace, NR. Phylogenetic stains: ribosomal RNA-based probes for the identification of single cells. Science. 1989 Mar 10;243(4896):1360–1363. [PubMed]
Felsenstein, J. Evolutionary trees from DNA sequences: a maximum likelihood approach. J Mol Evol. 1981;17(6):368–376. [PubMed]
A Unique Symbiosis in the Gut of Tropical Herbivorous Surgeonfish (Acanthuridae: Teleostei) from the Red Sea. Science. 1985 Jul 5;229(4708):49–51. [PubMed]
Ferguson, DJ; Birch-Andersen, A. Electron microscopy of a filamentous, segmented bacterium attached to the small intestine of mice from a laboratory animal colony in Denmark. Acta Pathol Microbiol Scand [B]. 1979 Aug;87(4):247–252. [PubMed]
Giovannoni, SJ; Britschgi, TB; Moyer, CL; Field, KG. Genetic diversity in Sargasso Sea bacterioplankton. Nature. 1990 May 3;345(6270):60–63. [PubMed]
Gunderson, JH; Sogin, ML; Wollett, G; Hollingdale, M; de la Cruz, VF; Waters, AP; McCutchan, TF. Structurally distinct, stage-specific ribosomes occur in Plasmodium. Science. 1987 Nov 13;238(4829):933–937. [PubMed]
Gutell, RR. Collection of small subunit (16S- and 16S-like) ribosomal RNA structures: 1994. Nucleic Acids Res. 1994 Sep;22(17):3502–3507. [PubMed]
Gutell, RR; Woese, CR. Higher order structural elements in ribosomal RNAs: pseudo-knots and the use of noncanonical pairs. Proc Natl Acad Sci U S A. 1990 Jan;87(2):663–667. [PubMed]
Johnson, JL; Francis, BS. Taxonomy of the Clostridia: ribosomal ribonucleic acid homologies among the species. J Gen Microbiol. 1975 Jun;88(2):229–244. [PubMed]
Kunstýr, I; Schiel, R; Kaup, FJ; Uhr, G; Kirchhoff, H. Giant gram-negative noncultivable endospore-forming bacteria in rodent intestines. Naturwissenschaften. 1988 Oct;75(10):525–527. [PubMed]
Losick, R; Youngman, P; Piggot, PJ. Genetics of endospore formation in Bacillus subtilis. Annu Rev Genet. 1986;20:625–669. [PubMed]
Magill, NG; Setlow, P. Properties of purified sporlets produced by spoII mutants of Bacillus subtilis. J Bacteriol. 1992 Dec;174(24):8148–8151. [PubMed]
Maidak, BL; Larsen, N; McCaughey, MJ; Overbeek, R; Olsen, GJ; Fogel, K; Blandy, J; Woese, CR. The Ribosomal Database Project. Nucleic Acids Res. 1994 Sep;22(17):3485–3487. [PubMed]
Olsen, GJ. Phylogenetic analysis using ribosomal RNA. Methods Enzymol. 1988;164:793–812. [PubMed]
Olsen, GJ; Matsuda, H; Hagstrom, R; Overbeek, R. fastDNAmL: a tool for construction of phylogenetic trees of DNA sequences using maximum likelihood. Comput Appl Biosci. 1994 Feb;10(1):41–48. [PubMed]
ROBINOW, CF. Observations on the structure of Bacillus spores. J Gen Microbiol. 1951 Aug;5(3):439–457. [PubMed]
ROBINOW, CF. Kurzer Hinweis auf Metabacterium polyspora. Z Tropenmed Parasitol. 1957 Mar;8(1-2):225–227. [PubMed]
Sanger, F; Nicklen, S; Coulson, AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. [PubMed]
Stünkel, S; Alves, J; Kunstýr, I. Characterization of two "Metabacterium" sp. from the gut of rodents. 2. Heteroxenic cultivation and proof of dipicolinic acid in "M. polyspora". Folia Microbiol (Praha). 1993;38(3):171–175. [PubMed]
Waterbury, JB; Stanier, RY. Patterns of growth and development in pleurocapsalean cyanobacteria. Microbiol Rev. 1978 Mar;42(1):2–44. [PubMed]
Winker, S; Woese, CR. A definition of the domains Archaea, Bacteria and Eucarya in terms of small subunit ribosomal RNA characteristics. Syst Appl Microbiol. 1991;14(4):305–310. [PubMed]
Woese, CR; Gutell, RR. Evidence for several higher order structural elements in ribosomal RNA. Proc Natl Acad Sci U S A. 1989 May;86(9):3119–3122. [PubMed]
Woese, CR; Kandler, O; Wheelis, ML. Towards a natural system of organisms: proposal for the domains Archaea, Bacteria, and Eucarya. Proc Natl Acad Sci U S A. 1990 Jun;87(12):4576–4579. [PubMed]
Woese, CR; Stackebrandt, E; Macke, TJ; Fox, GE. A phylogenetic definition of the major eubacterial taxa. Syst Appl Microbiol. 1985;6:143–151. [PubMed]
Woese, CR; Winker, S; Gutell, RR. Architecture of ribosomal RNA: constraints on the sequence of "tetra-loops". Proc Natl Acad Sci U S A. 1990 Nov;87(21):8467–8471. [PubMed]