Growth on sorbitol as sole carbon source Growth ability ofP. agglomeransstrains on sorbitol was studied using 200-μl microcultures in 100-well Bioscreen C MBR system honeycomb plates (well volume 400 μl) at 24°C with regular shaking at 15-min intervals in M9 minimal medium containing 10 mM sorbitol as sole carbon source. All strains were grown overnight in LB, collected
by centrifugation, and washed twice with sterile 0.9% NaCl before being inoculated in M9 at an initial OD600of about 0.02. Growth curves were measured in triplicates by periodically quantifying the absorbance through a 420- to 580-nm wide band filter (OD420-580 nm) using a Bioscreen C MBR system (Growth Curves Oy, Helsinki, Finland). Growth at 24°C and 37°C Growth ability of selectedP. agglomerans sensu strictostrains Selleck MK5108 was determined at 24°C and 37°C using the Bioscreen C MBR system. The protocol was the similar to that described above for growth on sorbitol, except selleck compound that LB medium was
used in place of minimal medium. The mean growth rate per hour (k) was calculated each 20 minutes Protein Tyrosine Kinase inhibitor according to the formula whereN 0andN t represent absorbance measured at two consecutive time points and Δtis the time interval (i.e., 1 h) between the two measurements. The highest optical density, the maximal growth rate, as well as the time needed to reach the latter value were recorded for each strain. A comparison of these parameters was performed among the average values obtained for clinical, biocontrol or plant-pathogenicP. agglomeransstrains. Correlations selleck screening library between OD420-580 nmmeasured in the Bioscreen C MBR system and number of colony forming unit (CFU) was estimated for representative strains by dilution plating on LB agar. Accession numbers The accession numbers for the sequences produced for this study are: 16S rRNA gene [GenBank: FJ611802-FJ611887];gyrBgene
[GenBank: FJ617346-FJ617427];hrcNgene [GenBank: FJ617428-FJ617436];pagRIgenes [GenBank: FJ656221-FJ656252]. With the exception ofpagRI, for which they are shown directly in the corresponding figure, accession numbers and other sources of reference sequences not obtained in this work are indicated below.Complete genomes:C. koseriATCC BAA-895 [NCBI: NC_009792],E. amylovoraEa273http://www.sanger.ac.uk/Projects/E_amylovora/,E. coliK-12 MG1655 [NCBI: NC_000913],Enterobactersp. 638 [NCBI: NC_009436],E. tasmaniensisEt1/99 [NCBI: NC_010694],K. pneumoniae342 [NCBI: NC_011283],P. stewartiisubsp.indologenesDC283http://www.hgsc.bcm.tmc.edu/microbial-detail.xsp?project_id=125.16S rRNA gene:E. cloacaeATCC 13047T[GenBank: AJ251469],E. sakazakiiATCC 51329 [GenBank: AY752937],Pantoea sp.LMG 2558 [GenBank: EF688010],Pantoea sp.LMG 2781 [GenBank: EU216736],Pantoea sp.LMG 24198 [GenBank: EF688009],Pantoea sp.LMG 24199 [GenBank: EF688012],Pantoea sp.