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1""}}JHTS01000055.1, range 27,755�C36,623) was analyzed for the presence of sigma housekeeping promoter sequence by three different programs: PromoterHunter9 (Klucar et al., 2010), Promoter prediction10 (Reese, 2001) and BPROM 11 (Solovyev and Salamov, 2011). As the PromoterHunter requires a weight matrix for the ?10 and ?35 sequences, such a matrix was created based on the P. aeruginosa promoters (Potvin et al., 2008), available at phiSITE12. The ARNold13 program was employed for the identification of rho-independent terminators (Lambert et al., 2004). Identification of prophages and genomic islands The PHAST14 software (Zhou et al., 2011; was applied to identify prophage sequences within the draft genome sequence of P482. IslandPick 3.015 (Dhillon et al., 2015) was used to screen Carboplatin the genome for the presence of genomic islands. As part of the IslandPick analysis, the contigs comprising the draft genome were aligned using the genome of P. entomophila L48T as a reference. L48T was chosen as the most closely related species for which a complete genomic sequence is available. Results Pseudomonas sp. P482 is a new representative of P. donghuensis Phylogenetic studies based on the analysis of 16S rRNA gene sequences (Figure ?(Figure1A)1A) and MLSA (Figure ?(Figure1B)1B) revealed that the closest relatives of strain P482 are P. donghuensis HYST, an isolate from lake water in China studied for high siderophore yield (Gao et al., 2015), P. vranovensis DSM 16006T, BI 6727 molecular weight a non-fluorescent pseudomonad obtained from soil next to a highway in Czech Republic (Tvrzov�� et al., 2006), and a well-studied insect pathogen, P. entomophila L48T (Mulet et al., 2012). The availability of genomic data for P482 and the three related strains enabled us to DZNeP investigate whether P482 belongs to one of these species by calculation of ANI-values for pairwise alignment of genomes (Konstantinidis and Tiedje, 2005; Goris et al., 2007; Richter and Rossell��-M��ra, 2009). The highest ANI-value of 99.24% was obtained for the comparison of genomes of strains P482 and HYST (Table ?(Table3).3). ANI-values calculated for the comparison of genome of P482 with the genomes of P. vranovensis DSM 16006T and P. entomophila L48T were 85.34 and 81.48%. respectively, thus much below the single species value. Considering that the postulated single species threshold is ANI ~ 95�C96%, the Pseudomonas sp. P482 should be classified as the same species as P. donghuensis. This species has recently been established by Gao et al. (2012), with strain HYST as a type strain and the only known representative. High similarity between P482 and HYST was also confirmed at the biochemical level using API 20NE and API 50CH (bioM��rieux, France; Table S6). A comparative genome analysis using EDGAR showed that the number of unique ORFs found for the P482 and HYST is 222 (4.3%) and 345 (6.5%), respectively. Some of the differences may have a reason since both genomes are in a draft format.