In short, the OM60 NOR5 isolates were obtained as fol lows. In October 2005 sediment samples have been collected from a tidal flat spot at Knigshafen bay, close to the town of Record for the German Island of Sylt. The approx. geo graphic coordinates with the sampling web-site have been 55. 04 North and eight. 42 East. Most samples have been obtained through the top rated oxic layer of muddy or sandy intertidal sedi ments. Just after transportation on the laboratory additional one.10 and 1.a hundred dilutions with the original sediment samples have been ready in artificial seawater, then 50 or 200 ul al iquots of every sample were spread on agar plates of Pla wealthy medium supplemented together with the antibiotics ampicil lin and cycloheximide additional within a concentration of 2. 0 g l every.
The precise composition of Pla rich medium has presently buy SCH 900776 been described elsewhere, essentially it truly is composed of artificial seawater supplemented with nutritional vitamins and trace elements that incorporates 0. 25 g l just about every of yeast extract, peptone and glucose as substrates. Colonies displaying a pinkish to red violet pigmentation appeared immediately after several weeks of incubation at 24 C. Pigmented colonies were more purified by subsequent transfers on Pla rich agar plates without having antibiotics. To determine pur ity as well as the phylogenetic affiliation of isolated strains the 16S rRNA genes had been PCR amplified from full cells then straight sequenced making use of an ABI 3130xl DNA se quencer, A complete of 240 red pigmented colonies were obtained, of which 22 may be affiliated towards the OM60 NOR5 clade by phylogenetic analyses of their partial 16S rRNA gene sequences.
Utilised strains, media ITF2357 and cultivation conditions Along with the newly isolated strains Rap1red and Ivo14T the next reference strains have been taken from your DSMZ culture collection and used for comparison. Chromatocurvus halotolerans DSM 23344T, Congregibacter litoralis DSM 17192T, Cro nobacter muytjensii DSM 21870T, Dasania marina DSM 21967T, Haliea mediterranea DSM 21924T, Haliea rubra DSM 19751T, Haliea salexigens DSM 19537T, Oceanicoccus sagamiensis DSM 25275T, Spongiibacter marinus DSM 17750T and Spongiibacter tropicus DSM 19543T, Haliea rubra CM41 15aT was de posited in the DSMZ by the Laboratoire Arago, Universit? Pierre et Marie Curie under the disorders of the Materials Transfer Agreement. The authen ticity on the used strains is confirmed by the Identi fication Service from the DSMZ by sequencing within the respective 16S rRNA genes. For schedule cultivation all strains were grown on Mar ine Broth or Agar 2216. The BChl a containing strains Ivo14T, DSM 17192T, DSM 19751T and DSM 23344T had been also grown inside a complex medium that was less nutrient rich and more suitable for your expression of photosynthetic pigments in these strains.