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Microbulbifer variabilis sp. nov. and Microbulbifer epialgicus sp. nov., isolated from Pacific marine algae, possess a rod–coccus cell cycle in association with the growth phase

¹ Marine Biotechnology Institute, Heita, Kamaishi 026-0001, Japan
² TechnoSuruga Laboratory Co. Ltd, Nagasaki, Shimizu-ku, Shizuoka 424-0065, Japan
³ Department of Fermentation Science, Faculty of Bio-Science, Tokyo University of Agriculture, Sakuragaoka, Setagaya-ku, Tokyo 156-8502, Japan

Correspondence
Miyuki Nishijima
mnishiji{at}tecsrg.co.jp

Phylogenetic and taxonomic characterization was performed for 14 strains of bacteria that produce anticancer antibiotics (pelagiomicins) (represented by strain Ni-2088^T) and one strain that produces UV-absorbing substances (strain F-104^T), isolated from marine algae and seagrass collected from coastal areas of tropical Pacific islands and a subtropical island of Japan. All 15 isolates were Gram-negative, strictly aerobic, non-motile and non-spore-forming. Sequence analysis of the 16S rRNA gene showed that the isolates occupied positions in the phylogenetic radiation of the genus Microbulbifer, with similarities of 93.6–97.6 %. The cells possessed a clearly discernible rod–coccus cell cycle in association with the growth phase; cells were rods during the growth phase and all converted to coccoid–ovoid cells when proliferation ceased. The coccoid–ovoid cells were optically denser than the rod cells and were viable for extended periods. They were considered to constitute a resting form. The type strains of described species of Microbulbifer were also found to possess identical rod–coccus cell cycles. The G+C content of the DNA was 48.1–49.7 mol%. The major respiratory quinone system was ubiquinone-8. The major fatty acids were C_{18 : 1}7c and C_{16 : 0}, and the hydroxy acids comprised C_{10 : 0} 3-OH, C_{12 : 0} 3-OH and iso-C_{11 : 0} 3-OH. The polar lipids comprised phosphatidylethanolamine, phosphatidylglycerol and phosphatidylserine. The group of 14 pelagiomicin-producing strains and strain F-104^T each constituted a single genomic species. Based on phylogenetic affiliation, phenotypic characteristics and genomic distinctness, the isolates represent two novel species in the genus Microbulbifer, for which the names Microbulbifer variabilis sp. nov. (type strain Ni-2088^T =MBIC01082^T =ATCC 700307^T) and Microbulbifer epialgicus sp. nov. (type strain F-104^T =MBIC03330^T =DSM 18651^T) are proposed.

The GenBank/EMBL/DDBJ accession numbers for the 16S rRNA gene sequences of strains Ni-2088^T, F-104^T and C-38 are AB167354, AB266054 and AB266055, respectively, that for the gyrB gene sequence of strain Ni-2088^T is AB276367 and those for the ftsZ gene sequences of strains Ni-2088^T, F-104^T, M. hydrolyticus ATCC 700072^T, M. salipaludis JCM 11542^T, M. elongatus DSM 6810^T and M. maritimus JCM 12187^T are AB371302, AB371303, AB371308, AB371307, AB371306 and AB371305, respectively.

Details of strain isolation, a neighbour-joining tree based on concatenated gyrB and ftsZ sequences, micrographs of cells of the novel strains and reference strains, fatty acid profiles and a dendrogram based on fatty acid data, 2D TLC of polar lipids of representative novel strains and reference strains and detailed DNA–DNA hybridization results are available as supplementary material with the online version of this paper.