Altererythrobacter Xiamenensis Sp. Nov., an Algicidal Bacterium Isolated from Red Tide Seawater

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Altererythrobacter Xiamenensis Sp. Nov., an Algicidal Bacterium Isolated from Red Tide Seawater International Journal of Systematic and Evolutionary Microbiology (2014), 64, 631–637 DOI 10.1099/ijs.0.057257-0 Altererythrobacter xiamenensis sp. nov., an algicidal bacterium isolated from red tide seawater Xueqian Lei,1,23 Yi Li,1,23 Zhangran Chen,1 Wei Zheng,1 Qiliang Lai,1,3 Huajun Zhang,1 Chengwei Guan,1 Guanjing Cai,1 Xujun Yang,1 Yun Tian1 and Tianling Zheng1,2 Correspondence 1State Key Laboratory of Marine Environmental Science and Key Laboratory of MOE for Coast and Tianling Zheng Wetland Ecosystems, School of Life Sciences, Xiamen University, Xiamen 361005, PR China [email protected] 2ShenZhen Research Institute of Xiamen University, ShenZhen, 518057, PR China 3Key Laboratory of Marine Biogenetic Resources, Third Institute of Oceanography, State Oceanic Administration, PR China A Gram-stain-negative, yellow-pigmented, aerobic bacterial strain, designated LY02T, was isolated from red tide seawater in Xiamen, Fujian Province, China. Growth was observed at temperatures from 4 to 44 6C, at salinities from 0 to 9 % and at pH from 6 to 10. Phylogenetic analysis based on 16S rRNA gene sequencing revealed that the isolate was a member of the genus Altererythrobacter, which belongs to the family Erythrobacteraceae. Strain LY02T was related most closely to Altererythrobacter marensis MSW-14T (97.2 % 16S rRNA gene sequence similarity), followed by Altererythrobacter ishigakiensis JPCCMB0017T (97.1 %), Altererythrobacter epoxidivorans JCS350T (97.1 %) and Altererythrobacter luteolus SW-109T (97.0 %). The dominant fatty acids were C18 : 1v7c,C17 : 1v6c and summed feature 3 (comprising T C16 : 1v7c and/or C16 : 1v6c). DNA–DNA hybridization showed that strain LY02 possessed low DNA–DNA relatedness to A. marensis MSW-14T, A. ishigakiensis JPCCMB0017T, A. T T epoxidivorans JCS350 and A. luteolus SW-109 (mean±SD of 33.2±1.3, 32.1±1.0, 26.7±0.7 and 25.2±1.1 %, respectively). The G+C content of the chromosomal DNA was 61.2 mol%. The predominant respiratory quinone was ubiquinone-10 (Q-10). According to its morphology, physiology, fatty acid composition and 16S rRNA gene sequence data, the novel strain most appropriately belongs to the genus Altererythrobacter, but can readily be distinguished from recognized species. The name Altererythrobacter xiamenensis sp. nov. is proposed (type strain LY02T5CGMCC 1.12494T5KCTC 32398T5NBRC 109638T). We have studied algicidal bacterial diversity associated with (Kumar et al., 2008), Altererythrobacter marinus (Lai et al., 2009), red tide algal blooms along the Xiamen coast of China and Altererythrobacter marensis (Seo & Lee, 2010), Altererythrobacter the research has revealed novel algicidial strains (Li et al., dongtanensis (Fan et al., 2011), Altererythrobacter namhicola and 2013). This study focused on one of these isolates, designated Altererythrobacter aestuarii (Park et al., 2011), Altererythrobacter LY02T, with algicidal activity against the harmful algal ishigakiensis (Matsumoto et al., 2011), Altererythrobacter species Alexandrium tamarense. Comparative 16S rRNA gene xinjiangensis (Xue et al., 2012), Altererythrobacter gangjinensis sequence analysis indicated that strain LY02T formed a clade (Jeong et al., 2013) and Altererythrobacter troitsensis within the genus Altererythrobacter.ThegenusAltererythrobacter (Nedashkovskaya et al., 2013). Accordingly, the aim of the (family Erythrobacteraceae, order Sphingomonadales, class present work was to determine the exact taxonomic position Alphaproteobacteria, phylum Proteobacteria) was created of strain LY02T by using polyphasic characterization by Kwon et al. (2007). At the time of writing, this including the determination of phenotypic properties and genus includes 12 species with validly published names: a detailed phylogenetic analysis based on 16S rRNA gene Altererythrobacter luteolus (Yoon et al., 2005), Altererythrobacter sequences. epoxidivorans (Kwon et al., 2007), Altererythrobacter indicus Strain LY02T was isolated from surface water samples collected in October 2012 at a depth of 1–2 m from red 3These authors contributed equally to this work. tide seawater in Xiamen (24u 339 N 118u 99 E), Fujian The GenBank/EMBL/DDBJ accession number for the 16S rRNA gene Province, China. The samples were serially diluted (10-fold sequence of strain LY02T is KC520828. dilution) in sterile seawater and 0.1 ml aliquots of each Downloaded from www.microbiologyresearch.org by 057257 G 2014 IUMS Printed in Great Britain 631 IP: 137.108.70.7 On: Mon, 04 Jul 2016 10:22:10 X. Lei and others 86 Erythrobacter aquimaris SW-110T (AY461441) 90 0.01 Erythrobacter longus DSM 6997T (AF465835) Erythrobacter nanhaisediminis T30T (FJ654473) Erythrobacter vulgaris 022-2 10T (AY706935) 58 Erythrobacter flavus SW-46T (AF500004) Erythrobacter gaetbuli SW-161T (AY562220) DSM 8509T (AB013354) 78 Erythrobacter litoralis 61 Erythromicrobium ramosum DSM 8510T (AF465837) UST081027-248T (HQ203045) 55 Erythrobacter pelagi Erythrobacter citreus RE35F/1T (AF118020) Erythrobacter seohaensis SW-135T (AY562219) Erythrobacter gangjinensis K7-2T (EU428782) Altererythrobacter aestuarii KYW147T (FJ997597) Altererythrobacter namhicola KYW48T (FJ935793) Novosphingobium indicum H25T (EF549586) Altererythrobacter indicus MSSRF26T (DQ399262) Novosphingobium tardaugens ARI-1T (AB070237) Altererythrobacter luteolus SW-109T (AY739662) 70 Altererythrobacter gangjinensis KJ7T (JF751048) Altererythrobacter xiamenensis LY02T (KC520828) Altererythrobacter ishigakiensis JPCCMB0017T (AB363004) Altererythrobacter epoxidivorans JCS350T (DQ304436) Altererythrobacter marensis MSW-14T (FM177586) Altererythrobacter marinus H32T (EU726272) Altererythrobacter xinjiangensis S3-63T (HM028673) 76 Altererythrobacter dongtanensis JM27T (GU166344) 98 Altererythrobacter troitsensis JCM 17037T (AY676115) Oceanibaculum indicum P24T (EU656113) Fig. 1. Neighbour-joining tree showing the phylogenetic positions of strain LY02T and representatives of some other related taxa, based on 16S rRNA gene sequences. Bootstrap values (expressed as percentages of 1000 replications) are shown at branch points; only values .50 % are shown. Oceanibaculum indicum P24T was used as the outgroup. Bar, 0.01 nt substitution rate (Knuc) units. dilution were spread onto marine agar 2216 (MA; Difco) The nearly full-length 16S rRNA gene sequence (1411 nt) of followed by incubation for 7 days at 28 uC. Individual strain LY02T was determined. Phylogenetic analysis of strain colonies of distinct morphology were further purified three LY02T based on the 16S rRNA gene sequence indicated that times and stored at 280 uC in marine broth 2216 (MB; this strain belonged to the family Erythrobacteraceae, Difco) supplemented with 10 % (v/v) glycerol. forming a robust clade within the genus Altererythrobacter (Fig. 1). Its closest relatives were A. marensis MSW-14T Genomic DNA was extracted according to the method of (97.2 % 16S rRNA gene sequence similarity), followed by A. Ausubel et al. (1995). The 16S rRNA gene sequence was ishigakiensis JPCCMB0017T (97.1 %), A. epoxidivorans amplified by PCR using primers 27F and 1492R (DeLong, JCS350T (97.1 %) and A. luteolus SW-109T (97.0 %). Strain 1992). Purification of the PCR product was carried out LY02T formed a separate phylogenetic clade with A. according to the protocol of the TIANquick midi purifica- ishigakiensis JPCCMB0017T. tion kit (Tiangen). The purified DNA was cloned into vector pMD19-T and sequenced. Sequences of related taxa were Cell morphology and motility were observed by using obtained from the GenBank database and EzTaxon-e server transmission electron microscopy (model JEM-2100HC; (http://eztaxon-e.ezbiocloud.net/) (Kim et al., 2012). JEOL) and phase-contrast light microscopy (model 50i; Phylogenetic analysis was performed using MEGA version 4 Nikon), with cells from the early exponential phase grown (Tamura et al., 2007) after multiple alignment of the data on MA at 28 uC. Colony morphology was examined from using DNAMAN (version 5.1). Evolutionary distances and cultures grown on MA for 2 days. The presence of clustering were determined by using the neighbour-joining flexirubin-type pigments was assessed using the bath- method (Saitou & Nei, 1987), and were evaluated by using ochromic shift test with 20 % KOH, as described by bootstrap values based on 1000 replications. Bernardet et al. (2002). The Gram reaction was determined Downloaded from www.microbiologyresearch.org by 632 International Journal of Systematic and Evolutionary Microbiology 64 IP: 137.108.70.7 On: Mon, 04 Jul 2016 10:22:10 Altererythrobacter xiamenensis sp. nov. Table 1. Characteristics that differentiate strain LY02T from closely related species of the genus Altererythrobacter Strains: 1, LY02T;2,A. luteolus SW-109T (Yoon et al., 2005); 3, A. marensis MSW-14T (Seo & Lee, 2010); 4, A. epoxidivorans JCS350T (Kwon et al., 2007); 5, A. ishigakiensis JPCCMB0017T (Matsumoto et al., 2011). Data for catalase, oxidase, API 20NE, API 20E and API ZYM for all five strains were done at the same time in this study. All the strains were positive for catalase activity, oxidase activity, and hydolysis of Tweens 20, 40 and 60 and tyrosine; negative for hydrolysis of chitin, gelatin and urea, and flexirubin-type pigments. In API 20NE strips, all strains were positive for b- glucosidase (aesculin hydrolysis); negative for indole production, arginine dihydrolase, gelatin hydrolysis, and utilization of L-arabinose, D- mannose, D-mannitol, N-acetylglucosamine, potassium gluconate, capric acid, trisodium citrate and phenylacetic acid. In API 20E
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