title: Three Novel Species with Peptidoglycan Cell Walls form the New GenusLacunisphaeragen. nov. in the Family Opitutaceae of the Verrucomicrobial Subdivision 4.
abstract: The cell wall of free-living bacteria consists of peptidoglycan (PG) and is critical for maintenance of shape as dissolved solutes cause osmotic pressure and challenge cell integrity. Surprisingly, the subdivision 4 of the phylum Verrucomicrobia appears to be exceptional in this respect. Organisms of this subdivision are described to be devoid of muramic or diaminopimelic acid (DAP), usually found as components of PG in bacterial cell walls. Here we describe three novel bacterial strains from a freshwater lake, IG15T, IG16bT, and IG31T, belonging to a new genus in the subdivision 4 of Verrucomicrobia which we found to possess PG as part of their cell walls. Biochemical analysis revealed the presence of DAP not only in these novel strains, but also inOpitutus terraePB90-1T, the closest described relative of strains IG15T, IG16bT, and IG31T. Furthermore, we found that nearly all genes necessary for peptidoglycan synthesis are present in genomes of subdivision 4 members, as well as in the complete genome sequence of strain IG16bT. In addition, we isolated and visualized PG-sacculi for strain IG16bT. Thus, our results challenge the concept of peptidoglycan-less free-living bacteria. Our polyphasic taxonomy approach places the novel strains in a new genus within the familyOpitutaceae, for which the nameLacunisphaeragen. nov. is proposed. Strain designations for IG15T, IG16bTand IG31TareLacunisphaera parvulasp. nov. (=DSM 26814 = LMG 29468),L. limnophilasp. nov. (=DSM 26815 = LMG 29469) andL. anatissp. nov. (=DSM 103142 = LMG 29578) respectively, withL. limnophilaIG16bTbeing the type species of the genus.
authors.affiliation: Microbial Cell Biology and Genetics, Leibniz-Institut DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH Braunschweig, Germany.
authors.forename: Patrick
authors.initials: P
authors.lastname: Rast
authors.affiliation: Institute for Pharmacology, Toxicology and Clinical Pharmacy, University of Technology Braunschweig, Germany.
authors.forename: Ines
authors.initials: I
authors.lastname: Glöckner
authors.affiliation: Microbial Cell Biology and Genetics, Leibniz-Institut DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH Braunschweig, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Boedeker
authors.affiliation: Microbial Cell Biology and Genetics, Leibniz-Institut DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH Braunschweig, Germany.
authors.forename: Olga
authors.initials: O
authors.lastname: Jeske
authors.affiliation: Microbial Cell Biology and Genetics, Leibniz-Institut DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH Braunschweig, Germany.
authors.forename: Sandra
authors.initials: S
authors.lastname: Wiegand
authors.affiliation: Max Planck Genome Center, Max Planck Institute for Plant Breeding Research Köln, Germany.
authors.forename: Richard
authors.initials: R
authors.lastname: Reinhardt
authors.affiliation: Department of Central Services, Leibniz-Institut DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH Braunschweig, Germany.
authors.forename: Peter
authors.initials: P
authors.lastname: Schumann
authors.affiliation: Central Facility for Microscopy, Helmholtz Centre for Infection Research Braunschweig, Germany.
authors.forename: Manfred
authors.initials: M
authors.lastname: Rohde
authors.affiliation: Department Microorganisms, Leibniz-Institut DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH Braunschweig, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Spring
authors.affiliation: Department of Molecular Ecology, Max Planck Institute for Marine Microbiology Bremen, Germany.
authors.forename: Frank O
authors.initials: FO
authors.lastname: Glöckner
authors.affiliation: Microbial Cell Biology and Genetics, Leibniz-Institut DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbHBraunschweig, Germany; Department of Microbiology, Institute for Water and Wetland Research, Faculty of Science, Radboud UniversityNijmegen, Netherlands.
authors.forename: Christian
authors.initials: C
authors.lastname: Jogler
authors.affiliation: Microbial Cell Biology and Genetics, Leibniz-Institut DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH Braunschweig, Germany.
authors.forename: Mareike
authors.initials: M
authors.lastname: Jogler
keywords: Lacunisphaera; Verrucomicrobia; ornithine; peptidoglycan; subdivision 4