Downloaded from www.microbiologyresearch.org by IP: 54.70.40.11 On: Thu, 06 Dec 2018 03:54:24 Nesterenkonia pannonica sp. nov., a novel alkaliphilic and moderately halophilic actinobacterium Andrea K. Borsodi, 1, * Tibor Szili-Kov acs, 2 Peter Schumann, 3 Cathrin Spröer, 3 K aroly M arialigeti 1 and Erika Tóth 1 Abstract An alkaliphilic and moderately halophilic bacterial strain characterized by optimal growth at pH 9.0–10.0 and with 5–7% (w/v) NaCl, designated BV-35 T , was isolated from water of a soda pan located in Kiskuns ag National Park, Hungary. Cells of the orange-pigmented colony were Gram-stain-positive, non-motile and non-endospore-forming coccoid rods. The isolate was strictly aerobic, catalase-positive and oxidase-negative. Strain BV-35 T displayed a peptidoglycan similar to type A4a, L-Lys–L-Glu (A11.54 according to www.peptidoglycan-types.info) but containing additionally 4-aminobutyric acid. Menaquinone-7 (MK-7) was the predominant isoprenoid quinone, and anteiso-C 15 : 0 and anteiso-C 17 : 0 were its major cellular fatty acids. The DNA G+C content of strain BV-35 T was 65.4 mol%. Based on 16S rRNA gene sequence similarities, the novel isolate showed the closest relationship to Nesterenkonia populi GP 10-3 T (97.9 %). The DNA–DNA relatedness between BV-35 T and N. populi was 46.7 %. The distinguishing phenotypic and genetic results of this polyphasic study revealed that strain BV-35 T represents a novel member of the genus Nesterenkonia, for which the name Nesterenkonia pannonica sp. nov. is proposed. The type strain is BV-35 T (=DSM 29786 T =NCAIM B 02606 T ). The genus Nesterenkonia was first described by Stacke- brandt et al. [1] following the taxonomic dissection of the genus Micrococcus into the genera Kocuria, Nesterenkonia, Kytococcus, Dermacoccus and Micrococcus. Subsequently, amendments for taxonomic description of the genus Nester- enkonia were provided by Collins et al. [2] and Li et al. [3] based on cell morphology, environmental tolerance, pepti- doglycan type and G+C content. The genus Nesterenkonia comprises Gram-positive, non-spore-forming, short-rod- shaped, aerobic, catalase-positive and oxidase-negative bacteria [1–3]. Nesterenkonia species have A4a-type pepti- doglycan with L-Lys–Gly–L-Glu, L-Lys–L-Glu or Lys–Gly–D- Asp, and their DNA G+C content varies between 64 and 72 mol% [3]. At present, the genus Nesterenkonia contains 17 moderately halophilic or halotolerant species, according to the List of Prokaryotes with Standing in Nomenclature (http://www. bacterio.net/). Type strains of Nesterenkonia species have been isolated mainly from saline or hypersaline and/or alka- line habitats such as solar saltern ponds [4], salt pans [5], hypersaline and soda lakes [2, 6], deep-sea sediments [7], saline and hypersaline soils [5, 8, 9], desert rhizosphere soils [3, 10] and Antarctic soil [11]. In addition, Nesterenkonia species have been described from fermented seafood [12], effluents of a paper mill [13], a cotton pulp mill [14], the faecal microbiota of an AIDS patient [15] and the bark of Populus euphratica [16]. The present study sought to determine the taxonomic posi- tion of strain BV-35 T isolated from water of a soda pan based on a polyphasic approach. The sample was collected from the water of the Böddi-sz ek soda pan (46  46¢ N 019  09¢ E) in Kiskuns ag National Park, Hungary. This shallow soda pan (average depth is less than 50 cm) has a fragmented water surface area with a maxi- mum of 1.88 km 2 . The alkaline lake water (pH 9.2) can be characterized by the dominance of Na + , HCO 3 – , CO 3 2 and Cl  ions [17]. The serially diluted water sample was spread on modified R2A medium (DSM medium no. 830) contain- ing 0.5 g yeast extract, 0.5 g proteose peptone (Difco no. 3), 0.5 g casamino acids, 0.5 g glucose, 0.5 g soluble starch, 0.3 g sodium pyruvate, 0.3 g K 2 HPO 4 , 0.05 g MgSO 4 .7H 2 O, 15.0 g agar and 1000.0 ml distilled water (www.dsmz.de) supple- mented with 5 % (w/v) NaCl. The pH of the modified R2A Author affiliations: 1 Department of Microbiology, ELTE Eötvös Lor and University, P azm any P. s et any 1/C, H-1117 Budapest, Hungary; 2 Institute for Soil Sciences and Agricultural Chemistry, Agricultural Research Center, Hungarian Academy of Sciences, Herman Ottó út 15, H-1022 Budapest, Hungary; 3 Leibniz Institute DSMZ – German Collection of Microorganisms and Cell Cultures, Inhoffenstrasse 7 B, 38124 Braunschweig, Germany. *Correspondence: Andrea K. Borsodi, borsodi.andrea@ttk.elte.hu Keywords: Nesterenkonia; alkaliphilic; moderately halophilic; soda pan. Abbreviations: GABA, 4-aminobutyric acid; MALDI-TOF, matrix-assisted laser desorption ionization time-of-flight; ML, maximum-likelihood; MP, maxi- mum-parsimony; NJ, neighbour-joining. The GenBank/EMBL/DDBJ accession number for the 16S rRNA gene sequence of strain BV-35 T is LN610504. One supplementary table and four supplementary figures are available with the online Supplementary Material. TAXONOMIC DESCRIPTION Borsodi et al., Int J Syst Evol Microbiol 2017;67:4116–4120 DOI 10.1099/ijsem.0.002263 002263 ã 2017 IUMS 4116