Photobacterium kishitanii sp nov., a luminous marine bacterium symbiotic with deep-sea fishes

Six representatives of a luminous bacterium commonly found in association with deep, cold-dwelling marine fishes were isolated from the light organs and skin of different fish species. These bacteria were Gram-negative, catalase-positive, and weakly oxiclase-positive or oxidase-negative. Morphologically, cells of these strains were coccoid or coccoid-rods, occurring singly or in pairs, and motile by means of polar flagellation. After growth on seawater-based agar medium at 22 degrees C for 18 h, colonies were small, round and white, with an intense cerulean blue luminescence. Analysis of 16S rRNA gene sequence similarity placed these bacteria in the genus Photobacterium. Phylogenetic analysis based on seven housekeeping gene sequences (16S rRNA gene, gapA, gyrB, pyrH, recA, rpoA and rpoD), seven gene sequences of the lux operon (luxC, luxD, luxA, luxB, luxF, luxE and luxG) and four gene sequences of the rib operon (ribE, ribB, ribH and ribA), resolved the six strains as members of the genus Photobacterium and as a clade distinct from other species of Photobacterium. These strains were most closely related to Photobacterium phosphoreum and Photobacterium iliopiscarium. DNA-DNA hybridization values between the designated type strain, Photobacterium kishitanii pjapo.1.1(T), and P. phosphoreum LMG 4233(T), p. iliopiscarium LMG 19543 T and Photobacterium indicum LMG 22857 T were 51, 43 and 19%, respectively. In AFLP analysis, the six strains clustered together, forming a group distinct from other analysed species. The fatty acid C-17:0 cyclo was present in these bacteria, but not in P. phosphoreum, P. iliopiscarium or P. indicum. A combination of biochemical tests (arginine clihydrolase and lysine clecarboxylase) differentiates these strains from P. phosphoreum and P. indicum. The DNA G+C content of P. kishitaniipjapo.1.1(T) is 40.2%, and the genome size is approximately 4.2 Mbp, in the form of two circular chromosomes. These strains represent a novel species, for which the name Photobacterium kishitanii sp. nov. is proposed. The type strain, pjapo. 1.1(T) (=ATCC BAA-1194(T) =LMG 23890(T)), is a luminous symbiont isolated from the light organ of the deep-water fish Physiculus japonicus.