Genomic insight into Chryseobacterium turcicum sp. nov. and Chryseobacterium muglaense sp. nov. isolated from farmed rainbow trout in Turkey

Four strains, designated as C-2, C-17^T, C-39^T and Ch-15, were isolated from farmed rainbow trout samples showing clinical signs during an investigation for a fish-health screening study. The pairwise 16S rRNA gene sequence analysis showed that strain C-17^T shared the highest identity level of 98.1 % with the type strain of Chryseobacterium piscium LMG 23089^T while strains C-2, C-39^T and Ch-15 were closely related to Chryseobacterium balustinum DSM 16775^T with an identity level of 99.3 %. A polyphasic approach involving phenotypic, chemotaxonomic and genome-based analyses was employed to determine the taxonomic provenance of the strains. The overall genome relatedness indices including dDDH and ANI analyses confirmed that strains C-2, C-17^T, C-39^T and Ch-15 formed two novel species within the genus Chryseobacterium. Chemotaxonomic analyses showed that strains C-17^T and C-39^T have typical characteristics of the genus Chryseobacterium by having phosphatidylethanolamine in their polar lipid profile, MK-6 as only isoprenoid quinone and the presence of iso-C_15:0 as major fatty acid. The genome size and G + C content of the strains ranged between 4.4 and 5.0 Mb and 33.5 – 33.6 %, respectively. Comprehensive genome analyses revealed that the strains have antimicrobial resistance genes, prophages and horizontally acquired genes in addition to secondary metabolite-coding gene clusters. In conclusion, based on the polyphasic analyses conducted on the present study, strains C-17^T and C-39^T are representatives of two novel species within the genus Chryseobacterium, for which the names Chryseobacterium turcicum sp. nov. and Chryseobacterium muglaense sp. nov. with the type strains C-17^T (=JCM 34190^T = KCTC 82250^T) and C-39^T (=JCM 34191^T = KCTC 822251^T), respectively, are proposed.     

Streptosporangium anatoliense sp. nov., isolated from soil in Turkey

A novel actinobacterium, strain N9999^T, was isolated from soil and its taxonomic position determined using a polyphasic approach. The organism formed abundant aerial hyphae that differentiated into spherical spore vesicles. The cell wall contained meso-diaminopimelic acid; the whole-cell sugars were galactose, glucose, mannose, madurose and ribose; the predominant menaquinones MK-9 (H₂) and MK-9 (H₄); the major phospholipids phosphatidylethanolamine, diphosphatidylglycerol, a phosphaglycolipid and phosphatidylinositol mannosides; while the cellular fatty acids were rich in iso-C_14:0, C_15:0, cis-9-C_17:1, iso-C_16:0 and 10-methyl C_17:0 components. Phylogenetic analyses based on an almost complete 16S rRNA gene sequence indicated that strain N9999^T was closely related to a group that consisted of Streptosporangium pseudovulgare DSM 43181^T and Streptosporangium nondiastaticum DSM 43848^T. However, DNA–DNA relatedness and phenotypic data demonstrated that strain N9999^T was clearly distinguished from all closely related Streptosporangium species. The combined genotypic and phenotypic data demonstrate conclusively that the isolate should be classified as a new species of Streptosporangium.     

Pseudomonas versuta sp. nov., isolated from Antarctic soil

In this study we analysed three bacterial strains coded L10.10^T, A4R1.5 and A4R1.12, isolated in the course of a study of quorum-quenching bacteria occurring in Antarctic soil. The 16S rRNA gene sequence was identical in the three strains and showed 99.7% pairwise similarity with respect to the closest related species Pseudomonas weihenstephanensis WS4993^T. Therefore, the three strains were classified within the genus Pseudomonas. Analysis of housekeeping genes (rpoB, rpoD and gyrB) sequences showed similarities of 84-95% with respect to the closest related species of Pseudomonas, confirming its phylogenetic affiliation. The ANI values were less than 86% to the closest related species type strains. The respiratory quinone is Q9. The major fatty acids are C16:0, C16:1 ω7c/ C16:1 ω6c in summed feature 3 and C18:1 ω7c / C18:1 ω6c in summed feature 8. The strains are oxidase- and catalase-positive. Growth occurs at 4–30 °C, and at pH 4.0–10. The DNA G+C content is 58.2–58.3 mol %. The combined genotypic, phenotypic and chemotaxonomic data support the classification of strains L10.10^T, A4R1.5 and A4R1.12 into a novel species of Pseudomonas, for which the name P. versuta sp. nov. is proposed. The type strain is L10.10^T (LMG 29628^T, DSM 101070^T).     

Bacillus daqingensis sp. nov. isolated from near poultry farm soil

A novel bacterial strain, designated PFS-5^T, was isolated from the soil environment with feces of a live poultry farm located in Cheonan, Republic of Korea. Strain PFS-5^T was Gram-staining-positive, motile, strictly aerobic bacterium, rod-shaped, and endospore-forming. The strain contained meso-diaminopimelic acid in their peptidoglycan and MK-7 menaquinone. The major fatty acids were anteiso-C_15:0 (44.2%), C_16:0 (22.2%), and iso-C_15:0 (16.7%). The DNA G+C content was 40.1 mol%. Comparative 16S rRNA gene sequence analysis identified strain PFS-5^T in the genus Bacillus, exhibiting the highest level of sequence similarity with type strain of B. herbersteinensis D-1,5a^T (96.9%), B. humi LMG 22167^T (96.7%), B. alkalitelluris BA288^T (96.1%), B. litoralis SW-211^T (96.0%), and B. luteolus YIM93174^T (95.5%). The major polar lipids of PFS-5^T were diphosphatidylglycerol and phosphatidylglycerol. On the basis of result from poly-phasic data, strain PFS-5^T represents a novel species, for which the name Bacillus cheonanensis sp. nov. is proposed (Type strai PFS-5^T = KACC 17469^T = JCM19333^T).     

Pseudomonas petrae sp. nov. isolated from regolith samples in Antarctica

A polyphasic taxonomic approach was used to characterize the four strains P2653^T, P2652, P2498, and P2647, isolated from Antarctic regolith samples. Initial genotype screening performed by PCR fingerprinting based on repetitive sequences showed that the isolates studied formed a coherent cluster separated from the other Pseudomonas species. Identification results based on 16S rRNA gene sequences showed the highest sequence similarity with Pseudomonas graminis (99.7%), which was confirmed by multilocus sequence analysis using the rpoB, rpoD, and gyrB genes. Genome sequence comparison of P2653^T with the most related P. graminis type strain DSM 11363^T revealed an average nucleotide identity of 92.1% and a digital DNA-DNA hybridization value of 46.6%. The major fatty acids for all Antarctic strains were C_16:0, Summed Feature 3 (C_16:1 ω7c/C_16:1 ω6c) and Summed Feature 8 (C_18:1 ω7c/C_18:1 ω6c). The predominant respiratory quinone was Q-9, and the major polar lipids were phosphatidylethanolamine, diphosphatidylglycerol, and phosphatidylglycerol. The regolith strains could be differentiated from related species by the absence of arginine dihydrolase, ornithine and lysine decarboxylase and by negative tyrosine hydrolysis. The results of this polyphasic study allowed the genotypic and phenotypic differentiation of four analysed strains from the closest related species, which confirmed that the strains represent a novel species within the genus Pseudomonas, for which the name Pseudomonas petrae sp. nov. is proposed with P2653^T (CCM 8850^T = DSM 112068^T = LMG 30619^T) as the type strain.     

Two novel species isolated from wheat rhizospheres in Serbia: Pseudomonas serbica sp. nov. and Pseudomonas serboccidentalis sp. nov.

Pseudomonas strains IT-194P, IT-215P, IT-P366^T and IT-P374^T were isolated from the rhizospheres of wheat grown in soils sampled from different fields (some of them known to be disease-suppressive) located near Mionica, Serbia. Phylogenetic analysis of the 16S rRNA genes and of whole genome sequences showed that these strains belong to two potentially new species, one containing strains IT-P366^T and IT-194P and clustering (whole genome analysis) next to P. umsongensis DSM16611^T, and another species containing strains IT-P374^T and IT-215P and clustering next to P. koreensis LMG21318^T. Genome analysis confirmed the proposition of novel species, as ANI was below the threshold of 95% and dDDH below 70% for strains IT-P366^T (compared with P. umsongensis DSM16611^T) and IT-P374^T (compared with P. koreensis LMG21318^T). Unlike P. umsongensis DSM16611^T, strains of P. serbica can grow on D-mannitol, but not on pectin, D-galacturonic acid, L-galactonic acid lactone and α-hydroxybutyric acid. In contrary to P. koreensis LMG21318^T, strains of P. serboccidentalis can use sucrose, inosine and α-ketoglutaric acid (but not L-histidine) as carbon sources. Altogether, these results indicate the existence of two novel species for which we propose the names Pseudomonas serbica sp. nov., with the type strain IT-P366^T (=CFBP 9060 ^T = LMG 32732 ^T = EML 1791 ^T) and Pseudomonas serboccidentalis sp. nov., with the type strain IT-P374^T (=CFBP 9061 ^T = LMG 32734 ^T = EML 1792 ^T). Strains from this study presented a set of phytobeneficial functions modulating plant hormonal balance, plant nutrition and plant protection, suggesting a potential as Plant Growth-Promoting Rhizobacteria (PGPR).     

New species of pathogenic Pseudomonas isolated from citrus in Tunisia: Proposal of Pseudomonas kairouanensis sp. nov. and Pseudomonas nabeulensis sp. nov.

A collection of Pseudomonas strains was isolated in different regions of Tunisia in the period 2016–2017 from the fruits and leaves of Citrus sinensis cv. ‘Valencia Late’ and Citrus limon cv. ‘Eureka’ plants with symptoms of blast and black pit disease. A phylogenetic analysis of the housekeeping gene rpoD was used for strain identification at the species level. The results demonstrated the affiliation of these strains with the genus Pseudomonas and revealed the presence of 11 strains representing two putative new species in two monophyletic branches. These strains were analyzed morphologically and genotypically by multilocus sequence analyses of the rpoD, gyrB and 16S rRNA (rrs) gene sequences, and their phenotypic characteristics by API 20NE and Biolog GEN III. Plant pathogenic properties were confirmed on fruits and detached leaves of C. limon cv. ‘Eureka’. Fatty acids and WC MALDI-TOF MS major protein profiles were determined. The genomes of both representatives were sequenced. The average nucleotide index and genome-to-genome distance from KC12^T and E10B^T are below the cut-off established for a described species. These results support the conclusion that the strains KC12^T, KC17, KC20, KC22, KC24A, KC25 and KC26 represent a novel species of Pseudomonas, for which the name of Pseudomonas kairouanensis is proposed. The type strain is KC12^T (=CECT9766 and CFBP 8662). The strains E10B^T, E10AB, E10CB1 and Iy3BA represent another novel species of Pseudomonas for which the name of Pseudomonas nabeulensis is proposed; the type strain is E10B^T (=CECT9765 and CFBP 8661).     

Geminicoccus roseus gen. nov., sp. nov., an aerobic phototrophic Alphaproteobacterium isolated from a marine aquaculture biofilter

A Gram-negative, strictly aerobic, diplococcoid bacterium (strain D2-3^T) was isolated from the biofilter of a recirculating marine aquaculture system. Phylogenetic analysis of the 16S rRNA gene sequence of D2-3^T indicated that the new organism occupied a novel lineage within the -1 subclass of Proteobacteria and was related to the genera Rhodothalassium, Azospirillum, Craurococcus, Acidiphilium, and Tistrella. The highest sequence similarity (90.8%) of the 16S rRNA gene sequence of D2-3^T was to that of Candidatus “Alysiosphaera europaea”. D2-3^T was mesophilic, heterotrophic, required sea salt, and had a pH optimum of 8.0. Growth in the presence of light resulted in the formation of pink colonies, a 25% increased cell yield, and a slightly increased growth rate. D2-3^T contained carotenoids and low amounts of bacteriochlorophyll a. Membranes of D2-3^T contained b-type cytochromes. The G+C content of the DNA was 60.3±0.1 mol%. Phylogenetic, morphological, physiological, and biochemical analyses demonstrated that D2-3^T represented a new aerobic phototrophic genus, for which the name Geminicoccus roseus gen. nov., sp. nov. is proposed for the type species (D2-3^T=DSM 18922^T=ATCC BAA-1445^T).     

Taxonomic study of bacteria isolated from natural mineral waters: proposal of Pseudomonas jessenii sp. nov. and Pseudomonas mandelii sp. nov

The taxonomic position of 23 strains isolated from mineral waters and previously grouped in the authentic pseudomonads on the basis of a phenotypic analysis (cluster IX, subclusters XIIIa and XIIIc of VERHILLE, S., ELOMARI, M., COROLER, L., IZARD, D., LECLERC, H. (Syst. Appl. Microbiol, 20, 137-149, 1997) has been genotypically further studied in the present work. On the basis of hybridization results, these strains were gathered into two new genomic groups for which we propose the names of Pseudomonas jessenii sp. nov. (Type strain CIP 105274) and Pseudomonas mandelii sp. nov. (Type strain CIP 105273). Deoxyribonucleic acid relatedness levels showed homologies ranging from 78 to 100% for Pseudomonas jessenii and from 77 to 100% for Pseudomonas mandelii. Furthermore, hybrization rates with 66 representative well characterized species or only partially characterized species of the genus Pseudomonas were below 53%, with delta Tm values of 7 degrees C and more. The mol% G + C content ranged from 57 to 58. The two new species presented basic morphological characteristics common to all pseudomonads. Various phenotypic features, such as denitrification, growth at 4 degrees C or 41 degrees C, trigonelline assimilation, alpha-L-glutamyl-L-histidine arylarmidase activity, growth on benzoate and meso-tartrate were found to differentiate Pseudomonas jessenii from Pseudomonas mandelii and from other Pseudomonas species. Pseudomonas jessenii encompassed a total of 9 strains from both phenotypic groups IX and XIIIa. Pseudomonas mandelii clustered a total of 13 strains from both phenotypic groups IX and XIIIc. Their clinical significance is unknown. The 16S rDNA of each type strain was sequenced and compared with the known sequences of the representative strains of the genus Pseudomonas. A phylogenetic tree was constructed to determine the intrageneric relationships within the genus Pseudomonas.     

Pseudomonas oryzae sp. nov. isolated from a paddy soil in South China

A Gram-staining-negative, rod-shaped and motile with several polar flagellums bacterium, designated WM-3^T, was isolated from a rice paddy soil in South China. Growth occurred with 0–3.0 % (w/v) NaCl (optimum 2.0 %), at pH 5.5–9.0 (optimum pH 7.0) and at 25–42 °C (optimum 30–37 °C) in liquid Reasoner’s 2A medium. Analysis of the 16S rRNA gene and gyrB gene sequences revealed that strain WM-3^T was most closely related to the type strains of the species Pseudomonas linyingensis and Pseudomonas sagittaria. Its sequence similarities with P. linyingensis CGMCC 1.10701^T and P. sagittaria JCM 18195^T were 97.4 and 97.3 %, respectively, for 16S rRNA gene, and were 94.1 and 94.2 %, respectively, for gyrB gene. DNA–DNA hybridization between strain WM-3^T and these two type strains showed relatedness of 35.6 and 30.9 %, respectively. G+C content of genomic DNA was 69.4 mol%. The whole-cell fatty acids mainly consisted of C_16:0 (30.0 %), C_16:1 ω6c and/or C_16:1 ω7c (19.3 %) and C_18:1 ω6c and/or C_18:1 ω7c (16.3 %). The results of phenotypic, chemotaxonomic and genotypic analyses clearly indicated that strain WM-3^T belongs to genus Pseudomonas but represents a novel species, for which the name Pseudomonas oryzae sp. nov. is proposed. The type strain is WM-3^T (=KCTC 32247^T =CGMCC 1.12417^T).     

Crocus demirizianus sp. nov. from northwestern Turkey

A new species, Crocus demirizianus O. Erol & L. Can is described and illustrated from northwestern Turkey. Crocus demirizianus was found in the Çanakkale province, near the district of Çan, and is undoubtedly related to the Crocus biflorus complex. However, the species differs from its relatives by typically sagittate anthers, and flowers without prominent stripes on outer segments. Diagnostic morphological characters are discussed and high resolution images are presented.     

Pseudomonas xanthomarina sp. nov., a novel bacterium isolated from marine ascidian

Two Pseudomonas-like yellow-orange-pigmented non-fluorescent denitrifying strains KMM 235 and KMM 1447T were isolated from marine ascidian specimens and investigated by a polyphasic approach to clarify their taxonomic status. On the basis of 16S rDNA gene sequence data the new isolates clustered with the Pseudomonas stutzeri species group with sequence similarities of >98%. The results of DNA-DNA hybridization and biochemical characterization showed genetic and phenotypic distinction between strains KMM 235 and KMM 1447T and from the other validly described Pseudomonas species. Strain KMM 235 was found to be closely related to the type strain of Pseudomonas stutzeri in their phenotypic and genetic characteristics and represented, probably, a new P. stutzeri genomovar. It is proposed that strain KMM 1447T be classified as a new species of the genus Pseudomonas, Pseudomonas xanthomarina sp. nov., with the type strain KMM 1447T (=JCM 12468T=NRIC 0617T=CCUG 46543T).     

Pseudomonas sihuiensis sp. nov., isolated from a forest soil in South China

A Gram-stain negative, motile, rod-shaped bacterium, designated strain WM-2^T, was isolated from a forest soil in Sihui City, South China, and characterized by means of a polyphasic approach. Growth occurred with 0–5 % (w/v) NaCl (optimum 0–1 %) and at pH 5.0–10.5 (optimum pH 8.5) and 4–40 °C (optimum 30 °C) in Luria–Bertani medium. Comparative 16S rRNA gene sequence analyses showed that strain WM-2^T is a member of the genus Pseudomonas and most closely related to P. guguanensis, P. oleovorans subsp. lubricantis, P. toyotomiensis, P. alcaliphila and P. mendocina with 97.1–96.6 % sequence similarities. In terms of gyrB and rpoB gene sequences, strain WM-2^T showed the highest similarity with the type strains of the species P. toyotomiensis and P. alcaliphila. The DNA–DNA relatedness values of strain WM-2^T with P. guguanensis and P. oleovorans subsp. lubricantis was 48.7 and 37.2 %, respectively. Chemotaxonomic characteristics (the main ubiquinone Q-9, major fatty acids C_18:1 ω7c/C_18:1 ω6c, C_16:0 and C_16:1 ω7c/C_16:1 ω6c and DNA G+C content 65.2 ± 0.7 mol%) were similar to those of members of the genus Pseudomonas. Polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, an unknown aminophospholipid, an unknown phospholipid and five unknown lipids. According to the results of polyphasic analyses, strain WM-2^T represents a novel species in the genus Pseudomonas, for which the name Pseudomonas sihuiensis sp. nov. is proposed. The type strain is WM-2^T (=KCTC 32246^T=CGMCC 1.12407^T).     

Thalassococcus lentus sp. nov., an alphaproteobacterium isolated from seawater of a seaweed farm

A Gram-negative, non-motile and rod- or ovoid-shaped bacterial strain, designated YCS-24^T, was isolated from seawater of a seaweed farm in the South Sea, South Korea. Strain YCS-24^T grew optimally at 25–28 °C, at pH 7.0–7.5 and in the presence of 2 % (w/v) NaCl. Strain YCS-24^T exhibited the highest 16S rRNA gene sequence similarity values of 97.5 and 97.1 % to the type strains of Thalassobius maritimus and Thalassococcus halodurans, respectively. The neighbour-joining, maximum-likelihood and maximum-parsimony phylogenetic trees based on 16S rRNA gene sequences revealed that strain YCS-24^T clustered with the type strain of T. halodurans. The DNA G+C content of strain YCS-24^T was 58.0 mol% and its DNA–DNA relatedness values with T. halodurans JCM 13833^T and T. maritimus GSW-M6^T were 17 ± 6.2 and 23 ± 9.2 %, respectively. The predominant ubiquinone found in strain YCS-24^T was Q-10 and the predominant fatty acid of strain YCS-24^T was C_18:1 ω7c. The major polar lipids of strain YCS-24^T were phosphatidylcholine, phosphatidylglycerol, one unidentified aminolipid and one unidentified lipid. Differential phenotypic properties, together with the phylogenetic and genetic distinctiveness, demonstrated that strain YCS-24^T is distinguished from T. halodurans. On the basis of the data presented, strain YCS-24^T (= KCTC 32084^T = CCUG 62791^T) represents a novel species of the genus Thalassococcus, for which the name Thalassococcus lentus sp. nov. is proposed.     

Ruegeria arenilitoris sp. nov., isolated from the seashore sand around a seaweed farm

A Gram-negative, motile and rod-shaped bacterial strain, G-M8^T, which was isolated from seashore sand around a seaweed farm at Geoje island in South Korea, was characterized taxonomically. It grew optimally at 30–37 °C, at pH 7.0–8.0 and in presence of 2 % (w/v) NaCl. A neighbour-joining phylogenetic tree based on 16S rRNA gene sequences revealed that strain G-M8^T joined the cluster comprising the type strains of Ruegeria atlantica and Ruegeria lacuscaerulensis, showing 97.5 % sequence similarity, by a bootstrap resampling value of 85.8 %. It exhibited 16S rRNA gene sequence similarity values of 95.4–96.7 % to the type strains of the other Ruegeria species. Strain G-M8^T exhibited the highest gyrB sequence similarity value (88.5 %) to the type strain of R. lacuscaerulensis. Strain G-M8^T contained Q-10 as the predominant ubiquinone and C_18:1 ω7c as the predominant fatty acid. The polar lipid profile of strain G-M8^T was similar to that of R. atlantica KCTC 12424^T. The DNA G+C content of strain G-M8^T was 64.6 mol% and its mean DNA–DNA relatedness values with R. atlantica KCTC 12424^T and R. lacuscaerulensis KCTC 2953^T were 18 ± 5.3 and 10 ± 3.6 %, respectively. Differential phenotypic properties, together with the phylogenetic and genetic distinctiveness, demonstrated that strain G-M8^T is distinguished from other Ruegeria species. On the basis of the data presented, strain G-M8^T (=KCTC 23960^T = CCUG 62412^T) represents a novel species of the genus Ruegeria, for which the name Ruegeria arenilitoris sp. nov. is proposed.     

Pseudomonas nitritireducens sp. nov., a nitrite reduction bacterium isolated from wheat soil

A Gram-negative, non-mobile, polar single flagellum, rod-shaped bacterium WZBFD3-5A2^T was isolated from a wheat soil subjected to herbicides for several years. Cells of strain WZBFD3-5A2^T grow optimally on Luria-Bertani agar medium at 30?°C in the presence of 0–4.0?% (w/v) NaCl and pH 8.0. 16S rRNA gene sequence analysis revealed that strain WZBFD3-5A2^T belongs to the genus Pseudomonas. Physiological and biochemical tests supported the phylogenetic affiliation. Strain WZBFD3-5A2^T is closely related to Pseudomonas nitroreducens IAM1439^T, sharing 99.7?% sequence similarity. DNA–DNA hybridization experiments between the two strains showed only moderate reassociation similarity (33.92?±?1.0?%). The DNA G+C content is 62.0?mol%. The predominant respiratory quinine is Q-9. The major cellular fatty acids present are C_16:0 (28.55?%), C_16:1ω6c or C_16:1ω7c (20.94?%), C_18:1ω7c (17.21?%) and C_18:0 (13.73?%). The isolate is distinguishable from other related members of the genus Pseudomonas on the basis of phenotypic and biochemical characteristics. From the genotypic, chemotaxonomic and phenotypic data, it is evident that strain WZBFD3-5A2^T represents a novel species of the genus Pseudomonas, for which the name Pseudomonas nitritereducens sp. nov. is proposed. The type strain is WZBFD3-5A2^T (=CGMCC 1.10702^T?=?LMG 25966^T).