Pseudomonas wadenswilerensis sp nov and Pseudomonas reidholzensis sp nov., two novel species within the Pseudomonas putida group isolated from forest soil

Within the frame of a biotechnological screening, we isolated two Pseudomonas strains from forest soil. 16S rRNA gene sequence analysis indicated that strain CCOS 864(T) shared 99.8% similarity with Pseudomonas donghuensis HYST, while strain CCOS 865(T) shared 99.0% similarity with Pseudomonas putida DSM 291(T) and lower similarity with other P. putida group type strains. Based on multilocus sequence analysis, the two strains were genotypically distinct from each other, each forming a separate clade. Strains CCOS 864(T) and CCOS 865(T) were Gram-stain-negative, motile and rod-shaped, growing at a temperature range of 4-37 degrees C. Strain CCOS 864(T) could be phenotypically distinguished from P. putida group species by the combination of gelatinase-positive reaction and positive growth on N-acetyl-D-glucosamine, p-hydroxyphenylacetic acid and inosine but lack of fluorescein production on King's B medium, while strain CCOS 865(T) could be distinguished from P. putida group species by the combination of positive growth with saccharic acid and negative growth with p-hydroxyphenylacetic acid and L-pyroglutamic acid. The major polar lipid for both strains was phosphatidylethanolamine; the major quinone was ubiquinone Q-9. DNA-DNA hybridization and average nucleotide identities confirmed the novel species status for the two strains. The DNA G+C contents of CCOS 864(T) and CCOS 865(T) were 62.1 and 63.8 mol%, respectively. The phenotypic, phylogenetic and DNA-DNA relatedness data support the suggestion that CCOS 864(T) and CCOS 865(T) represent two novel Pseudomonas species. The names Pseudomonas wadenswilerensis sp. nov. (type strain CCOS 864(T) = LMG 29327(T)) and Pseudomonas reidholzensis sp. nov. (type strain CCOS 865(T) = LMG 29328(T)) are proposed.