Peña et al. Standards in Genomic Sciences (2016) 11:55 DOI 10.1186/s40793-016-0178-2 EXTENDED GENOME REPORT Open Access High quality draft genome sequences of Pseudomonas fulva DSM 17717T, Pseudomonas parafulva DSM 17004T and Pseudomonas cremoricolorata DSM 17059T type strains Arantxa Peña1, Antonio Busquets1, Margarita Gomila1, Magdalena Mulet1, Rosa M. Gomila2, T. B. K. Reddy3, Marcel Huntemann3, Amrita Pati3, Natalia Ivanova3, Victor Markowitz3, Elena García-Valdés1,4, Markus Göker5, Tanja Woyke3, Hans-Peter Klenk6, Nikos Kyrpides3,7 and Jorge Lalucat1,4* Abstract Pseudomonas has the highest number of species out of any genus of Gram-negative bacteria and is phylogenetically divided into several groups. The Pseudomonas putida phylogenetic branch includes at least 13 species of environmental and industrial interest, plant-associated bacteria, insect pathogens, and even some members that have been found in clinical specimens. In the context of the Genomic Encyclopedia of Bacteria and Archaea project, we present the permanent, high-quality draft genomes of the type strains of 3 taxonomically and ecologically closely related species in the Pseudomonas putida phylogenetic branch: Pseudomonas fulva DSM 17717T, Pseudomonas parafulva DSM 17004T and Pseudomonas cremoricolorata DSM 17059T.Allthreegenomesarecomparableinsize(4.6–4.9 Mb), with 4,119–4,459 protein-coding genes. Average nucleotide identity based on BLAST comparisons and digital genome-to- genome distance calculations are in good agreement with experimental DNA-DNA hybridization results. The genome sequences presented here will be very helpful in elucidating the taxonomy, phylogeny and evolution of the Pseudomonas putida species complex. Keywords: Genomic Encyclopedia of Type Strains (GEBA), One Thousand Microbial Genomes Project (KMG-I), P. fulva, P. parafulva, P. cremoricolorata, Genome, Type strains Abbreviations: ANIb, Average nucleotide identity based on BLAST; dDDH, Digital DNA-DNA hybridization; DDH, DNA- DNA hybridization; gANI, Whole-genome ANI-based MiSI method; GEBA, Genomic encyclopedia of Bacteria and Archaea; KMG, One thousand microbial genomes; MIGS, Minimum information about a genome sequence; MLSA, Multilocus sequence analysis; NAS, Non-traceable; TAS, Traceable. * Correspondence: [email protected] 1Department of Biology-Microbiology, Universitat de les Illes Balears, Campus UIB, Crtra. Valldemossa km 7.5, 07122 Palma de Mallorca, Spain 4Institut Mediterrani d’Estudis Avançats (IMEDEA, CSIC-UIB), Palma de Mallorca, Spain Full list of author information is available at the end of the article © 2016 The Author(s). Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Peña et al. Standards in Genomic Sciences (2016) 11:55 Page 2 of 10 Introduction DSM 17004T (=AJ 2129T =JCM 11244T =NRIC 0501T) During a taxonomic study of Pseudomonas strains iso- and P. cremoricolorata DSM 17059T (=JCM 11246T lated from rice, petroleum fields and oil-brine in Japan, =NRIC 0181T), were obtained from the DSMZ. All Iizuka and Komagata [1] proposed two new species in strains were isolated by Iizuka and Komagata [1, 2] from 1963, Pseudomonas fulva and Pseudomonas straminea Japanese rice paddies and were initially proposed as (as cited in Uchino et al. [2]). These new species produced members of the new species P. fulva due to the deep yel- a water-insoluble yellow pigment, but not a water-soluble low color of their colonies. P. fulva was included in the fluorescentpigment.Later,sevenP. fulva strains obtained Approved Lists of Bacterial Names [12]. Uchino and col- from culture collections were re-characterized and com- laborators re-characterized several strains obtained as P. pared with the strains of related species by Uchino and col- fulva from culture collections and proposed two new laborators [2]. Phylogenetic analysis based on 16S rRNA species: P. parafulva (2 strains) and P. cremoricolorata sequences, experimental DNA-DNA hybridization results (1 strain) [2]. and phenotypic characteristics led to the proposal of two All three type strains shared the basic phenotypic traits new species: Pseudomonas parafulva (2 strains) and of the genus Pseudomonas: Gram-negative rods, motility Pseudomonas cremoricolorata (1 strain). Three of the four via polar flagella, with strictly respiratory type of metab- remaining strains were maintained in the species P. fulva, olism, catalase and oxidase activity and phylogenetic and the other strain was identified as P. straminaea [2]. In placement in the genus Pseudomonas on the basis of amultilocussequenceanalysis,thetypestrainsofP. fulva, 16S rRNA gene sequencing. None of the three species P. parafulva and P. cremoricolorata clustered in the produced water-soluble fluorescent pigments but pro- Pseudomonas putida phylogenetic branch and are consid- duced a characteristic water-insoluble yellow pigment. ered members of the P. putida group in the Pseudomonas Colonies appear smooth, round, flat to convex and pale/ fluorescens lineage [3]. The three species are taxonomically creamy yellow on nutrient agar. The three species were and ecologically closely related. Strains from these species differentiated from each other by several phenotypic tests: have been isolated from rice paddy samples or from presence of the arginine dihydrolase pathway, growth at Japanese unhulled rice. P. fulva strains have also been stud- 37 °C and assimilation of D-ribose, D-mannose, adonitol, ied for their endophytic growth in Scots pines and for their 2-keto-D-gluconate, butyrate, valerate, caprate, isovalerate, roles in plant growth promotion and protection against itaconate, citraconate, glycerate, levulinate, Tween 80, p- plant pathogenic fungi [4, 5]. The antagonistic effect against hydroxybenzoate, inosine, glycine, L-ornithine, L-citrulline plant pathogenic bacteria has also been demonstrated in and nicotinate. An extensive list of phenotypic characteris- other strains of Pseudomonas putida [6]. Additionally, P. tics can be found in the original publication by Uchino et fulva strains have been isolated from water collected from al. [2]. The classification and general features of P. fulva, human-made container habitats of mosquitoes [7]. P. fulva P. parafulva and P. cremoricolorata type strains are given was one of the most abundant species found in a survey of in Tables 1, 2 and 3. pseudomonads in human homes [8], and very recently P. fulva was identified as a member of a polymicrobial ventri- Chemotaxonomic data culitis in humans [9]. The difficulty in identifying species As reported by Uchino and collaborators [2] the DNA closely related to P. putida in the clinical laboratory is GC-content of the three type strains, as determined by highlighted by Rebolledo and collaborators [9]. Biosynthesis chemical analysis, was 60.0 mol % in P. fulva and P. of medium-chain-length poly(3-hydroxyalkanoates) by a parafulva and 62.1 mol % in P. cremoricolorata.The volatile aromatic hydrocarbons-degrading P. fulva has been percentages of G + C bases based on the genome analysis proposed as candidate for the biotechnological conversion were 61.71 % for P. fulva DSM 17717T, 62.42 % for P. of toxic petrochemical wastes to valuable biopolymers [10]. parafulva DSM 17004T and 63.47 % for P. cremoricolor- In the context of the Genomic Encyclopedia of Bacteria ata DSM 17059T. The GC-contents determined by and Archaea (GEBA) project [11], the permanent, high chemical analysis were slightly lower than those inferred quality draft genomes of the type strains of P. fulva, P. from genome sequences. The predominant respiratory parafulva and P. cremoricolorata are presented. The gen- quinone was ubiquinone Q-9, but Q-8 and Q-10 were ome sequences have been annotated, and the results are also detected in smaller amounts. The major cellular discussed in relation to the taxonomy of members of the fatty acids were C16:0, C16:1 and C18:1, and the major P. putida phylogenetic group. 3-hydroxy fatty acids were C10:0 and C12:0 [2]. For protein analysis, cells were cultured in Luria- Organisms information Bertani broth aerobically, with shaking at 30 °C, har- Classification and features vested in the exponential growth phase and prepared for The type strains of the three species, P. fulva DSM Whole-cell MALDI-TOF MS analysis using an Autoflex 17717T (=JCM 11242T =NRIC 0180T), P. parafulva III mass spectrometer (Bruker Daltonik, Germany) as Peña et al. Standards in Genomic Sciences (2016) 11:55 Page 3 of 10 Table 1 Classification and general features of P. fulva DSM Table 2 Classification and general features of P. parafulva DSM 17717T [36] 17004T [36] MIGS ID Property Term Evidence MIGS ID Property Term Evidence code code Classification Domain Bacteria TAS [37] Classification Domain Bacteria TAS [37] Phylum Proteobacteria TAS [38] Phylum Proteobacteria TAS [38] Class TAS [39] Class Gammaproteobacteria TAS [39] Gammaproteobacteria Order Pseudomonadales TAS [40] Order Pseudomonadales TAS [40] Family Pseudomonadaceae TAS [41] Family TAS [41] Pseudomonadaceae Genus Pseudomonas TAS