Downloaded from www.microbiologyresearch.org by IP: 131.204.246.172 On: Wed, 07 Sep 2016 13:21:19 2784 gene rRNA AP-38 16S strain the of sequence for number accession GenBank/EMBL/DDBJ The genus The Microbiology Evolutionary and Systematic of Journal International nanjingensis genus variabile crobium the to assigned al et he upeetyfgrsaeaalbewt h nieSupplementary online the with available Material. are figures supplementry Three halotolerans genus the ing, ulse names: published speioachiayiensis (Yoon 2004), giessen.de [email protected] K Peter Correspondence 20)t comdt nayia pce formerly atypical an accommodate to (2004) . tal et speioahypogeus Isoptericola ampfer € Isoptericola ,2006), ., (Huang (Zhang Isoptericola (Bakalidou speioavariabilis Isoptericola speioajiangsuensis Isoptericola tal et T tal et a ntal rpsdb Stackebrandt by proposed initially was sKU201961. is (Tseng Cellulosimicrobium ,21) w ute pce have species further Two 2012). ., nldssvnseiswt validly with species seven includes ,2005), ., (Groth tal et tal et ,20) ttetm fwrit- of time the At 2002). ., rmsanpstv,arbcogns,soiga reua elmrhlg,wsioae from isolated ( was cucumber morphology, of cell tissue irregular root an the showing organism, aerobic Gram-stain-positive, A diinlypoiepeoyi ifrnito fsri AP-38 strain of differentiation phenotypic provide additionally oiin ntebsso h 6 RAgn euneaayi,sri AP-38 strain analysis, sequence gene rRNA 16S the of basis the On position. igotcdaioai fteppiolcnwas peptidoglycan the of acid diamino diagnostic to related closely most cucumis Isoptericola h yestrain. type the anteiso- profile, acid fatty the In tyramine. of amounts C significant components and major spermine the and contained spermidine pattern polyamine The two mannosides. and phosphatidylinositol phosphatidylinositol phosphatidylglycerol, diphosphatidylglycerol, were compounds 4 3 2 1 Hu, Chia-Hui ( cucumber of tissue root h 6 RAgn eunesmlrt oalohrseiso h genus the of species other all to similarity sequence gene rRNA 16S The rps h name the propose nanjingensis tant h genus the to strain rdmnnl eaunnsMK-9(H menaquinones predominantly  nanjingensis ee K Peter ntttfrMkoilge Veterin Mikrobiologie, für D- Institut Hans-Knöll-Institut-, V., e. Germany Jena, Infektionsbiologie USA 07745 und 36849, Naturstoff-Forschung Alabama für University, Auburn Leibniz-Institut Pathology, Plant and Entomology of Department Justus-Liebig-Universit Germany Mikrobiologie, Angewandte für Institut ,21)and 2011) ., 5:0 : 15 85% DNA %. 98.5 tal et speioadokdonensis Isoptericola (Stackebrandt n iso-C and ,2005), ., (Wu ampfer, € as AP-38 . S 24300 DSM tal et – Cellulosimi- Isoptericola Isoptericola N eaens to relatedness DNA 5:0 : 15 2 speioacucumis Isoptericola ,2010), ., onA McInroy, A. John Isoptericola 1 T eepeeti ao mut.Teedt upr h loaino the of allocation the support data These amounts. major in present were tal et ersnsanvlseiso h genus the of species novel a represents tfneP Glaeser, P. Stefanie speioavariabilis Isoptericola (2016), T ., a 1rcpoa 1% n 4(eircl3 ) epciey The respectively. %), 34 (reciprocal 34 and %) 41 31(reciprocal was uui sativus Cucumis h eut fpyilgcladbohmclcharacterization biochemical and physiological of results The . 66, reiiiceUniversit armedizinische € ‘ atra tanAP-38 strain bacterial enpooe u hi ae aentytbe validly been yet not have names their published, but proposed been xi) elmrhlgclfaue eercre with 28 recorded at TSA were on features grown cells morphological Cell Oxoid). oeie 3 sometimes ae eemndb sn noiaets (Merck). test oxidase an using by by determined described dase, as (analysed Gerhardt Gram-positive weakly stained ufc fe 8ha 28 at smooth h a 48 with after colonies some- yellow surface rod-shaped, forming to cells coccoid irregular time single, produced strain The ht rmtehatyitra ottsu fcucumber of tissue root internal healthy ( the from phyte uigepnnilgot,irglrysae el fstrain of cells AP-38 shaped irregularly growth, exponential During speioarhizophila Isoptericola uui sativus Cucumis 2784 speioavariablis Isoptericola 4 n MK-9(H and ) p o. ihAP-38 with nov., sp. – 2 T p o. sltdfo the from isolated nov., sp. 2788 ai Martin Karin eeosre,0.9 observed, were tal. et 9. )and %) (99.1 n netgtdi ealfristaxonomic its for detail in investigated and ) ‘ speioasalitolerans Isoptericola – 1 uui sativus Cucumis mln,wihsoe omtlt.Cells motility. no showed which long, µm 4 94 n eengtv o yohm oxi- cytochome for negative were and 1994) , L oehW Kloepper, W. Joseph lsn.Teqioesse contained system quinone The -lysine. ,wsinvestigated. was ), 001055 2 .I h oa ii rfl,major profile, lipid polar the In ). t -20We,Austria Wien, A-1210 at, € T ’ S 10177 DSM ioae nteery19sa nendo- an as 1990s early the in ,isolated speioananjingensis Isoptericola (Kaur 3 T ã T n asJre Busse Hans-Jürgen and tGesn -59 Giessen, D-35392 Giessen, at €   =LG29223 LMG (= ntytn o gr(TSA; agar soy tryptone on C from ypaecnrs microscopy. contrast phase by C Isoptericola 06IUMS 2016 – . mwd n 1.5 and wide µm 1.5 tal et .variabilis I. DOI ,21) nti td,the study, this In 2014). ., T ’ and (Guan 10.1099/ijsem.0.001055 Isoptericola o hc we which for , Isoptericola rne nGetBritain Great in Printed 2 T T CM8653 =CCM and a hw obe to shown was tal et ) I. 9. %). (98.9 ,21)and 2013) ., was – . µm 2.5 T as ) 4 Downloaded from www.microbiologyresearch.org by IP: 131.204.246.172 On: Wed, 07 Sep 2016 13:21:19 ehd mxmmlklho,maximum-parsimony), (maximum-likelihood, methods genus the of species monophyletic all a But, of Material). cluster Supplementary online the jingensis 02 Guan 2012; other all of genus proposals the the of in con- shown species was analysis This the method. with sistent neighbour-joining the by obtained ehdapidwt h yesrisof strains type the with applied method h 6 RAgn fsri AP-38 strain of gene rRNA 16S The Mpopaebfe dutdt h aep) both pH); same the to 28 of adjusted at addition cultured buffer the were by the phosphate by stabilized mM units and pH NaOH 5 0.5 or in HCl val- (increasing of pH 10 to addition and sup- adjusted 4 or either pH NaCl between Difco) (w/v) ues % (TSB; 10 to broth 1 soy with plemented tryptic in analysed was ocuin nteaayi efre ihteneighbour- the (Groth with method performed joining analysis the on conclusions http://ijs.microbiologyresearch.org 25, 15, 4, 42 at and TSA 37 on 32, growth 28, after investigated was Temperature-dependent growth observed. be not could Endospores eunesmlrt a bandto obtained Highest strains. was type similarity related database closely sequence sequence most the gene to similarities rRNA 16S (Kim strain type EzTaxon tanAP-38 Strain Felsenstein, analysis; (bootstrap repetitions 1985). 100 and 1978) e pniggn emn 8 nucleoti- 1432 termini of gene size spanning a des had sequence final The pherograms. ae sn R ees . (Ludwig 5.2 release ARB using gated The 5 1991). unclear removing (Lane, by 1492R 3 manually and and corrected 8F was sequence primers with analysis universal phylogenetic the for sequenced subsequently and the tal et aaaerlae13o uy21.Tesqec a aligned was sequence The 2015. using July of 123 release database ‘ ae n1SrN eesqecsbtengn termini gene to between were (according sequences 1440 trees using gene and 96 All rRNA method 16S 2005). on (Felsenstein, based maximum-parsimony 3.6 version the v7.04 DNAPARs and RAxML bootstrap rapid analysis, and using GTR-GAMMA with method 2006) (Stamatakis, maximum-likelihood neighbour-joining) the (ARB Jukes the with method using calculated neighbour-joining were trees the Phylogenetic family analysis. the sequence of strains type all including alignment sequence al et SILVA h hlgntcpaeeto tanAP-38 strain of placement phylogenetic The pce ftegenus the of species and l-pce iigTree Living All-Species ,20;Wu 2006; ., ,20)adipeetdit h T aaae The database. LTP the into implemented and 2007) ., .nanjingensis I. rrnB ¢ tal et SINA euneed n ydtie oto fteelectro- the of control detailed by and ends sequence edainet(tp/wwabslad;Pruesse (http://www.arb-silva.de; alignment seed and ,21)wsue odtriepiws sequence pairwise determine to used was 2014) ., of v..;Pruesse (v1.2.9; tal et .variabilis I. shrci coli Escherichia T – tal., et atrcreto Jks&Cno,1969), Cantor, & (Jukes correction Cantor lsee neednl ftetreeing the of independently clustered ,21;Kaur, 2013; .,  9. ) eunesmlrt oalother all to similarity Sequence %). (98.9  .Slnt-adp-eedn growth pH-dependent and Salinity- C. Isoptericola C. Isoptericola, 00 Tseng 2010; tal et ’ Fg ,adS n 2aalbein available S2 and S1 and 1, (Figs rjc LP (Yarza (LTP) Project a hce aulybefore manually checked was .coli E. ,20;Zhang 2005; ., – tal et 44[ubrdacrigto according [numbered 1474 (Brosius was tal et ,21)acrigt the to according 2012) ., ubrn;Brosius numbering; hc lasbsdtheir based always which tal et  ,21) te treeing Other 2014). ., 85%. 98.5 T Isoptericola .variabilis I. tal et ,21;Huang 2011; ., tal et a PCR-amplified was .hypogeus I. tal et ,20)adthe and 2004) ., ,17).The 1978)]. ., T ,20;Yoon 2005; ., tal et a investi- was a only was 9. %) (99.1 , ,2008) ., .nan- I. tal et tal et ., ., ¢ hc otie 66%mnqioeMK-9(H menaquinone MK-9(H % 46.6 contained which oe n oa iiswshretda h stationary by Altenburger the reported and as at (1988) Auling extracted harvested & were was Busse Polyamines qui- lipids phase. acids, polar growth diamino of exponential and (1988), extraction Auling late nones for & use the Busse biomass by at whereas recommended as harvested phase was growth 28 from at peptone biomass 7.2) % pH (0.3 analysis, extract, broth yeast PYE % 0.3 in casein, grown and was quinones polyamines, lipids of polar analyses to subjected Biomass (Figs genus the of S2). and monophyly S1 the support not did however, ru.Teplaieptencnand[nµo gdry (g µmol [in contained phosphate pattern a or polyamine group The amino an group. did residue, that sugar detected a were contain not lipids polar three Additionally, manno- sides. phosphatidylinositol two phosphatidylglycerol, and phosphatidylinositol diphosphatidylglycerol, lipids major ye sdsrbdb idl 19a )adAltenburger and b) Stolz (1990a, Tindall al ana- by and et described extracted were as lipids Busse lysed polar by and described Quinones conditions (1997). HPLC using analysed eto tanAP-38 strain of ment 1. Fig. MK-8(H h aefml,the family, same the Isoptericola a ae nncetd eune pnig1SrN gene rRNA 16S spanning 96 sequences positions nucleotide sequence on based was rtdin erated otta values Bootstrap ubro pce hc r nlddi h lse.Outgroups of the cluster. sequences represent the strain in cluster type included a the are within were which Numbers species values of bootstrap analyses. number high other with represent the supported circles also by Larger were S2). that and nodes those S1 max- (Figs and maximum-parsimony trees the imum-likelihood in present also nodes represent uloieposition. nucleotide phragmiteti Cellulomonas 0.01 19) h PCaprtsue a ecie by described was used apparatus HPLC The (1996). . tal et egbu-onn resoigtepyoeei place- phylogenetic the showing tree Neighbour-joining ARB 2 2 .Teplrlpdpoie(i.S)cnitdo the of consisted S3) (Fig. profile lipid polar The ). MK-9(H % 13.5 ), 20) tanAP-38 Strain (2007). . 84 n yesriso pce falgnr eogn to belonging genera all of species of strains type and Cellulomonas flavigena 83 sn ihtenibu-onn ehd h analysis The method. neigbour-joining the with using 80 81 3 9 75  Isoptericola cucumis 0%aegvna oe.Fle ice tnodes at circles Filled nodes. at given are % 70 Isoptericola nanjingensis Cellulomonas phragmiteti Isoptericola dokdonensis Isoptericola jiangsuensis ‘ Isoptericola salitolerans Promicromonosporaceae Cellulosimicrobium Isoptericola halotolerans ‘ Xylanimicrobium pachnodae Isoptericola rhizophila T Isoptericola hypogeus Isoptericola chiayiensis – ihtp tan fseiso h genus the of species of strains type with 4 Xylanibacterium ulmi 40(codn to (according 1440 Xylanimonas cellulosilytica a,00 uloiesbtttosper substitutions nucleotide 0.01 Bar, . Promicromonospora 6 ,38%MK-8(H % 3.8 ), NCIMB 8073 T Myceligenerans MX5 AP-38 ellmnsflavigena Cellulomonas hwdaqioesystem quinone a showed speioacucumis Isoptericola H17 ’ BKS3-46 (KC608148) HKI 0342 T ’ TRM F109 (JX453452) CLG DS-3 (AJ298873) YIM 70177 YIM KB23 XIL08 T 06182M-1 (KU201961) T T (X79463) (HQ222356) T VPCX2 T  (EU852101) h rewsgen- was tree The . (DQ387860) T .Frpolyamine For C. .coli E. tal et T (AM902253) (AY273185) XIL07 T (AJ854061) T

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D- – N 4 -inositol, rmti td.Dt o tan 4 strains for Data study. this from 3 n MK-9(H and ) sorbitol, -acetyl- tal. et D- antl uctl salicin, dulcitol, mannitol, D- 21) tan:1 AP-38 1, Strains: (2012). mannose, D T -cellobiose, D- =M 29223 (=LMG CLG D L- aatsmn,ribose, galactosamine, mnio,lcoe afns,tre- raffinose, lactose, -mannitol, 5:0 : 15 arabinose, T 6, ; 2 n anteiso-C and ,wt oeaeaonsof amounts moderate with ), D À À À À À À À L- -arabinose, .dokdonensis I. D- hmoe urs,meli- sucrose, rhamnose, obtlaentutilized. not are sorbitol D- galactose, L- T hmoe maltose, rhamnose, CM8653 =CCM T L- 2, ; N D- 5:0 : 15 À À À À DS-3 + + + + + + + yie h acyl the lysine; 4 -acetyl- rbtl Sev- arabitol. n MK-8 and ) – D- .nanjingensis I. rmHuang from 7 h diag- The . D- D- T gluconate, T 7, ; adonitol, adonitol, D- n h otcoeyrltdtp tan ftegenus the of strains type related closely most the and T xylose, .halotolerans I. D- ,was ), glu- À À À À + + + + + + + + + + + S 24300 DSM tal. et 21) aafrsris2ad3wr xcl ncnrec ihthose with congruence in exactly were 3 and 2 strains for Data (2012). I 70177 YIM nenlro iseo uubr( cucumber of tissue root internal lt uloiesqec fa1SrbsmlRAgn rmEscherichia from gene (1978). RNA F. ribosomal H. 16S Noller, a coli. & of J. sequence P. nucleotide Kennedy, plete L., M. Palmer, J., Brosius, eimfo h idu ftetrieMsoemsdarwiniensis. Mastotermes Microbiol termite Evol the of hindgut the from terium (2002). H. König, aaio,A,K A., Bakalidou, a n pce ftegnr rvbceim oyeatru,adtsuka- and corynebacterium, brevibacterium, genera murella. the of species and can (1997). lebre,P,K P., Altenburger, References for Sowinsky wall. cell Maria analyses the HPLC of and the acids for amino Grebing Bardl the Bettina of Katja and assistance Will, technical excellent Gundula thank We Acknowledgements USA. AL, Auburn, University, Auburn at greenhouse a Biotechnol J (1996). J. lebre,P,K P., Altenburger, rcNt cdSiUSA S U Sci Acad Natl Proc T À À À À À À À À À À À À À À À À À À À À À À À À W (=H17 oymn itiuini cioyee ihgopBpeptidogly- B group with actinomycetes in distribution Polyamine n ytBacteriol Syst J Int lsiiaino atraioae rmamdea alpainting. wall medieval a from isolated of Classification T 47 8, ; 39 , 52 T ;3, ); 1185 , .chiayiensis I. mfr . ectl,M,Khik . ezl .& M. Wenzel, T., Kuhnigk, M., Berchtold, P., ampfer, € – ellsmcoimvraies.nv,acluoyi bac- cellulolytic a nov., sp. variabile Cellulosimicrobium 52. mfr . arsahs . uiz .&Bse H.- Busse, & W. Lubitz, A., Makristathis, P., ampfer, € mfr . kmv .N,Lbt,W us,H.-J. Busse, & W. Lubitz, N., V. Akimov, P., ampfer, € .variabilis I. À À À À À À À À À À À À À À À À W + + + + + + + + – 1192. 47 75 270 , 06182M-1 4801 , S 10177 DSM – 277. – À À À À À À À À À À À À À À À À + + + 4805. 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