title: A culture-dependent bacterial community structure analysis based on liquid cultivation and its application to a marine environment.
abstract: A method for analyzing culture-dependent bacterial community structure by liquid cultivation was established using 96-well microplates. Using 96-well microplates, this method can easily provide accurate enumeration of viable microorganisms and simultaneous separation of bacteria, which allowed us to analyze the bacterial community. Bacteria in diluted surface seawater were separated using 96-well microplates and cultivated with 1/5 ZoBell 2216E liquid medium. The 98 cultures obtained were subsequently applied to phylogenetic analysis based on 16S rRNA gene sequences. The bacterial diversity, evaluated by the Shannon-Weaver index, was relatively small but comparable to previously reported bacterial communities of several environments. The most abundant group was the family Rhodobacteraceae, which has been frequently detected in marine environments. Most bacteria were phylogenetically related to bacteria or uncultured clones detected in marine environments, but distant from published species. The analysis of bacterial community structure by liquid cultivation would be useful as an alternative culture-dependent approach.
authors.affiliation: Department of Food Science, Faculty of Applied Life Sciences, Niigata University of Pharmacy and Applied Life Sciences, Niigata, Japan. shige@nupals.ac.jp
authors.forename: Toru
authors.initials: T
authors.lastname: Shigematsu
authors.forename: Mayumi
authors.initials: M
authors.lastname: Hayashi
authors.forename: Isamu
authors.initials: I
authors.lastname: Kikuchi
authors.forename: Shigeaki
authors.initials: S
authors.lastname: Ueno
authors.forename: Haruhiko
authors.initials: H
authors.lastname: Masaki
authors.forename: Tomoyuki
authors.initials: T
authors.lastname: Fujii
mesh_terms: D001419:Bacteria / Q000145:classification* / Q000235:genetics / Q000254:growth & development* / Q000302:isolation & purification; D044822:Biodiversity; D016000:Cluster Analysis; D003470:Culture Media; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D010802:Phylogeny; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D041902:Rhodobacteraceae / Q000145:classification / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification; D012623:Seawater / Q000382:microbiology*; D017422:Sequence Analysis, DNA
chemical_list: D003470:Culture Media; D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S