Downloaded from www.microbiologyresearch.org by IP: 203.232.191.197 On: Thu, 28 Jun 2018 07:40:54 Soonwooa purpurea sp. nov., isolated from a fresh water river Mercy Rose Stella Sirra, 1 † Shalem Raj Padakandla 1 † and Jong-Chan Chae 1,2, * Abstract A pale-brown-coloured, rod-shaped, non-motile, and Gram-reaction-negative bacterium, strain I54 T , was isolated from a water sample of a freshwater river in Iksan, Republic of Korea. The phylogenetic affiliation based on 16S rRNA gene sequence analysis showed that strain I54 T belonged to the genus Soonwooa of the family Flavobacteriaceae with a sequence similarity of 97.5 % to Soonwooa buanensis HM0024 T . The major fatty acids (>5 %) of strain I54 T were iso-C 15 : 0 , anteiso-C 15 : 0 , summed feature 3 (C 16 : 1 !7c/C 16 : 1 !6c) and iso-C 17 : 0 3-OH. Phosphatidylethanolamine, one aminolipid and two unknown lipids were the major polar lipids. The G+C content of the genomic DNA was 34.2 (±0.3)mol% and MK6 was the sole respiratory quinone. On the basis of its molecular and phenotypic characteristics, strain I54 T represents a novel species in the genus Soonwooa, for which the name Soonwooa purpurea sp. nov. is proposed. The type strain is I54 T (=KCTC 52722 T =JCM 31880 T ). The genus Soonwooa in the family Flavobacteriaceae was proposed by Joung et al. [1] based on the description of strain HM0024 T isolated from coastal seawater of the Yellow Sea, Republic of Korea. Ever since that description, there have been no reports on the occurrence of strains belonging to the genus Soonwooa except for two strains iso- lated from the Nile tilapia Oreochromis niloticus [2] and two strains from refrigerated bone-in whole raw duck meat [3]. In this communication, we present the isolation and polyphasic characterization of a Gram-reaction-negative, aerobic, rod-shaped, non-motile and non-spore-forming bacterium, and its description as the member of a novel spe- cies within the genus Soonwooa. Strain I54 T was isolated from water sample of Iksan river, Republic of Korea as a chloramphenicol-resistant bacte- rium. Pure culture of the strain was obtained by repeated streaking on Mueller–Hinton agar plates containing chlor- amphenicol (16 μg ml 1 ) and was preserved as stocks in each of glycerol (20 %), sucrose (10 %) and skim milk (10 %) at 80  C. Following the protocol described previously [4], identification of strain I54 T was carried out by PCR amplifi- cation and sequencing of the 16S rRNA gene using universal primers 27F (5¢-AGAGTTTGATCMTGGCTCAG-3¢) and 1492R (5¢- TACGGYTACCTTGTTACGACTT-3¢). Com- parison of the almost-complete sequence (1418 bp) with the available 16S rRNA gene sequences using EzBioCloud server [5] revealed that strain I54 T was phylogenetically close to Soonwooa buanensis HM0024 T with a sequence similarity of 97.5 % followed by less than 95.0 % with the members of the genera Elizabethkingia and Chryseobacte- rium in the family Flavobacteriaceae. By using the neighbour-joining (NJ) [6], maximum-parsimony (MP) [7] and minimum evolution (ME) [8] methods in MEGA6 soft- ware [9], phylogenetic analysis of the 16S rRNA gene sequences of strain I54 T and those of the most closely related type strains belonging to the genera Elizabethkingia and Chryseobacterium was performed. Evolutionary distan- ces in the NJ and ME trees were calculated in a pair-wise deletion procedure using Kimura’s two-parameter correc- tion [10]. Robustness of the phylogenetic trees was evalu- ated by a bootstrap procedure with 1000 replications [11]. All of the three phylogenetic trees summarized in Fig. 1 showed that strain I54 T clustered with S. buanensis HM0024 T with high bootstrap support. For taxonomic analysis, the type strain S. buanensis HM0024 T (=KCTC 22689 T ) was procured from the Korean Collection for Type Cultures (KCTC). As strain I54 T showed optimum growth on tryptone soy agar (TSA) and the type strain S. buanensis HM0024 T was also reported to grow on TSA [1], further comparative characterization was carried on TSA/tryptone soy broth (TSB) following the recommended standards pro- posed by Tindall et al. [12] and Bernardet et al. [13]. Author affiliations: 1 Division of Biotechnology, Chonbuk National University, Iksan 54596, Republic of Korea; 2 Advanced Institute of Environment and Bioscience, Chonbuk National University, Iksan 54596, Republic of Korea. *Correspondence: Jong-Chan Chae, chae@jbnu.ac.kr Keywords: Soonwooa purpurea; polyphasic characterization. Abbreviations: AL, aminolipid; APL, aminophospholipid; JCM, Japan Collection of Microorganisms; KCTC, Korean Collection for Type Cultures; PE, phosphatidylethanolamine; TSA, tryptone soy agar; TSB, tryptone soy broth; UL, unknown lipid. †These authors contributed equally to this work. The GenBank/EMBL/DDBJ accession number for the 16S rRNA gene sequence of strain I54 T is LT717348. One supplementary table and three supplementary figures are available with the online version of this article. TAXONOMIC DESCRIPTION Sirra et al., Int J Syst Evol Microbiol 2017;67:5193–5197 DOI 10.1099/ijsem.0.002441 002441 ã 2017 IUMS 5193