description of galbitalea soli gen. nov., sp. nov., and frondihabitans sucicola sp. nov

Description of Galbitalea soli gen. nov., sp. nov.,and Frondihabitans sucicola sp. nov.

Soo-Jin Kim,1 Jun-Muk Lim,1,2 Jae-Hyung Ahn,1 Hang-Yeon Weon,1

Moriyuki Hamada,3 Ken-ichiro Suzuki,3 Tae-Young Ahn2

and Soon-Wo Kwon1

Correspondence

Soon-Wo Kwon

[email protected]

1Agricultural Microbiology Division, National Academy of Agricultural Science, Rural DevelopmentAdministration, Suwon 441-707, Republic of Korea

2Department of Microbiology, Dankook University, Anseo-dong, Cheonan 330-714, Republic of Korea

3Biological Resource Center (NBRC), National Institute of Technology and Evaluation,2-5-8 Kazusakamatari, Kisarazu, Chiba 292-0818, Japan

Bacterial strains KIS82-1T and GRS42T were isolated from soil and from sap of Acer mono,

respectively, in the Republic of Korea. Both strains were aerobic, Gram-stain-positive, mesophilic,

rod-shaped and motile. Phylogenetically, both strains belonged to the family Microbacteriaceae of

the phylum Actinobacteria. The 16S rRNA gene sequence of strain KIS82-1T showed the highest

similarity to those of Frondihabitans peucedani RS-15T (97.6 %), Frigoribacterium mesophilum

MSL-08T (97.2 %) and Labedella gwakjiensis KSW2-17T (97.0 %), while strain GRS42T showed

the highest 16S rRNA gene sequence similarity to Frondihabitans peucedani RS-15T (98.7 %),

Frondihabitans cladoniiphilus CafT13T (98.4 %), Frondihabitans australicus E1HC-02T (98.2 %)

and Frigoribacterium faeni 801T (97.3 %). The 16S rRNA gene sequence similarity between

GRS42T and KIS82-1T was 97.0 %. Phylogenetic trees indicated that strain GRS42T was firmly

grouped into the genus Frondihabitans, while strain KIS82-1T did not show a clear affiliation to

any genus within the family Microbacteriaceae. Strain KIS82-1T showed type B1b peptidoglycan

with 2,4-diamino-L-butyric acid as the diamino acid. It had MK-11, MK-10 and MK-12 as

respiratory quinones, anteiso-C15 : 0, iso-C16 : 0 and iso-C14 : 0 as major cellular fatty acids and

diphosphatidylglycerol, phosphatidylglycerol and an unknown glycolipid as predominant polar

lipids. The peptidoglycan of strain GRS42T was of type B2b with D-ornithine as the diamino acid.

The strain contained MK-8, MK-9 and MK-7 as respiratory quinones, summed feature 8

(C18 : 1v6c and/or C18 : 1v7c) as major cellular fatty acid and diphosphatidylglycerol, phospha-

tidylglycerol and three unknown glycolipids as predominant polar lipids. Strain GRS42T revealed

low DNA–DNA hybridization (,50 % relatedness) with closely related strains. Based on the data

obtained in the present polyphasic taxonomic study, we propose that strain KIS82-1T represents

a novel genus and species and that strain GRS42T represents a novel species in the family

Microbacteriaceae. The genus Galbitalea gen. nov. is proposed, with strain KIS82-1T (5KACC

15520T5NBRC 108727T) as the type strain of the type species, Galbitalea soli sp. nov. Strain

GRS42T (5KACC 15521T5NBRC 108728T) is proposed as the type strain of Frondihabitans

sucicola sp. nov.

The family Microbacteriaceae was proposed by Park et al.(1993), and its description was emended by Stackebrandtet al. (1997). Members of the family Microbacteriaceae have

been found in diverse environments including plants, soil,air, dairy products, sewage, mushrooms, compost, insectsand groundwater. At the time of writing, 40 genera wererecognized within the family Microbacteriaceae (http://www.bacterio.net/classifgenerafamilies.html). Recently, the newgenera Alpinimonas, Diaminobutyricimonas, Lysinimonas,Naasia and Pontimonas, isolated from air samples, seawaterand silt, have been reported as new members of the familyMicrobacteriaceae (Jang et al., 2012, 2013a, b; Schumannet al., 2012; Weon et al., 2013). We isolated two strains,

Abbreviation: L-DAB, 2,4-diamino-L-butyric acid.

The GenBank/EMBL/DDBJ accession numbers for the 16S rRNA genesequences of strains KIS82-1T and GRS42T are JX876866 andJX876867, respectively.

Three supplementary figures are available with the online version of thispaper.

International Journal of Systematic and Evolutionary Microbiology (2014), 64, 572–578 DOI 10.1099/ijs.0.058339-0

572 058339 G 2014 IUMS Printed in Great Britain

KIS82-1T and GRS42T, from soil and from sap of the treeAcer mono, respectively. According to our phylogeneticanalysis, these strains are newly identified members of thefamily Microbacteriaceae. Strain GRS42T formed a clusterwith the members of the genus Frondihabitans; however,strain KIS82-1T was not clearly grouped into any genuswithin the family.

Strain KIS82-1T was isolated from a soil on Kaui island,Taean county, Republic of Korea. The soil was suspendedin 0.85 % NaCl (w/v), serially diluted and then spread ontoR2A medium. The plates were incubated at 28 uC for6 days, and strain KIS82-1T was purified. Strain GRS42T

was isolated from sap of Acer mono growing on BaikamMountain, Hwasun county, Republic of Korea. StrainGRS42T was isolated by the above-mentioned method.

16S rRNA genes were amplified by using the universalprimers 9F and 1512R (Weisburg et al., 1991), and thepurified PCR products were sequenced by Solgent (Daejeon,Republic of Korea). The almost-complete 16S rRNA genesof strains KIS82-1T and GRS42T were 1467 and 1474 bp,respectively. Searches for phylogenetic neighbours based on16S rRNA gene sequence similarity were performed usingthe EzTaxon-e server (http://eztaxon-e.ezbiocloud.net/; Kimet al., 2012). Sequence alignment and analysis of the datawere performed using the ARB software package (versionDecember 2007; Ludwig et al., 2004) and the correspondingSILVA SSURef 100 database (release August 2009; Pruesseet al., 2007). Phylogenetic trees were reconstructed usingMEGA version 5.0 (Tamura et al., 2011) on the basis ofthe neighbour-joining (Saitou & Nei, 1987), maximum-parsimony (Kluge & Farris, 1969) and maximum-likelihood(Felsenstein, 1981) algorithms. The 16S rRNA genesequence of strain KIS82-1T showed the highest similarityto those of Frondihabitans peucedani RS-15T (97.6 %),Frigoribacterium mesophilum MSL-08T (97.2 %) andLabedella gwakjiensis KSW2-17T (97.0 %), showing similar-ity levels below 97.0 % to members of other species withinthe family Microbacteriaceae with validly published names.Strain GRS42T showed the highest 16S rRNA gene sequencesimilarity to Frondihabitans peucedani RS-15T (98.7 %), Fron-dihabitans cladoniiphilus CafT13T (98.4 %), Frondihabitansaustralicus E1HC-02T (98.2 %) and Frigoribacterium faeni801T (97.3 %), revealing less than 96.5 % 16S rRNA genesequence similarity to members of other species of the familyMicrobacteriaceae. The 16S rRNA gene sequence similaritybetween GRS42T and KIS82-1T was 97.0 %. Phylogenetically,strain KIS82-1T was clustered with Frigoribacterium mesophilumMSL-08T, but this grouping was not fully supported bybootstrap values (Figs 1 and S1, available in IJSEM Online).Other trees reconstructed using the maximum-parsimony andmaximum-likelihood algorithms showed that strain KIS82-1T

clustered with members of the genera Lysinimonas andCryobacterium, or the genus Frondihabitans (data not shown).These unstable tree topologies made the phylogenetic posi-tion of strain KIS82-1T within any genus of the familyMicrobacteriaceae unclear. In the neighbour-joining phylogen-etic tree, strain GRS42T formed a cluster with members of the

genus Frondihabitans with high bootstrap support (85 %) (Fig.1). This was also supported by the maximum-parsimony andmaximum-likelihood trees (Fig. 1) and the expanded neigh-bour-joining tree (Fig. S1).

The cell morphology and motility of strains KIS82-1T andGRS42T were examined by light microscopy (AXIO; Zeiss)and transmission electron microscopy (LEO model912AB), with cells grown on R2A agar at 28 uC for 2 days.For transmission electron microscopy, cells were negativelystained with 0.5 % (w/v) uranyl acetate. Growth waschecked on nutrient agar (NA; BD), trypticase soy agar(TSA; BD), International Streptomyces Project (ISP)medium 2 (Shirling & Gottlieb, 1966) and MacConkeyagar (BD). Growth conditions were checked by culturing atdifferent temperatures, pH and NaCl concentrations for upto 14 days. Growth was monitored using R2A broth at 4,10, 15, 20, 25, 28, 30, 33, 35, 37, 40 and 45 uC. The pHrange (pH 3.0–11.0 at intervals of 1.0 pH units) for growthwas determined in R2A broth that was buffered with 0.2 Mcitrate/phosphate buffer, 0.05 M Tris/HCl, HCl or NaOH(Breznak & Costilow, 1994). NaCl tolerance was tested bygrowth in R2A broth containing 0, 1, 2, 3, 5 or 7 % (w/v)NaCl. Gram staining was performed using Hucker’smethod (Smibert & Krieg, 1994). Catalase and oxidaseactivities were examined by bubble production in 3 % (v/v)hydrogen peroxide solution and using 1 % (w/v) tetra-methyl p-phenylenediamine (bioMerieux), respectively.Degradation of casein, starch and tyrosine was examinedon R2A plates containing milk powder (5 %, w/v), starch(1 %, w/v) and L-tyrosine (0.1 %, w/v), respectively.Degradation of CM-cellulose and Tween 80 was examinedusing R2A supplemented with 1 % (w/v) of each substrate.DNase activity was determined with DNase test agar (BD).Anaerobic growth was determined by incubation in theBBL GasPak Anaerobic System (BD) for 14 days at 28 uCon R2A agar. Enzyme activities, utilization of variouscarbon sources and acid production from substrates weretested in duplicate with commercial API ZYM, API 20NEand API ID 32GN kits (bioMerieux) according to themanufacturer’s protocols.

Both strains KIS82-1T and GRS42T were Gram-stain-positive,motile rods (Fig. S2). Strain KIS82-1T was catalase- andoxidase-negative, grew at 10–37 uC (optimum, 28–30 uC)and at pH 4.0–9.0 (optimum, pH 7.0) and tolerated 1 % butnot 2 % (w/v) NaCl. It grew on NA and TSA, but not on ISPmedium 2 or MacConkey agar. Strain GRS42T was catalase-positive and oxidase-negative, grew at 4–30 uC (optimum,28 uC) and pH 4.0–9.0 (optimum, 7.0) and tolerated 2 % butnot 3 % (w/v) NaCl. It grew on NA, TSA, ISP medium 2 andMacConkey agar.

For cellular fatty acid analysis, cells were grown on R2A at28 uC for 24 or 48 h, and reached the exponential stage ofgrowth. Cellular fatty acids were extracted, methylated andseparated by gas chromatography (model 6890; HewlettPackard) according to the protocol of the SherlockMicrobial Identification System (MIDI; Sasser, 1990).

Two novel taxa in the family Microbacteriaceae

http://ijs.sgmjournals.org 573

Fatty acid methyl esters were identified and quantifiedusing the TSBA 6 database (version 6.10) of the SherlockMicrobial Identification System (MIDI). Menaquinonesand polar lipids were extracted and analysed using cellsgrown on R2A agar at 28 uC for 48 h by the method ofMinnikin et al. (1984). For detection of polar lipids, TLCplates were sprayed with molybdatophosphoric acid (fortotal lipids), phosphomolybdic acid (for phospholipids),ninhydrin (for aminolipids) or a-naphthol/sulfuric acid(for glycolipids). Peptidoglycan analysis was conducted asdescribed by Schleifer & Kandler (1972). Strains KIS82-1T

and GRS42T were grown in liquid YG medium (1.0 % yeastextract, 1.0 % glucose; pH 7.3) on a rotary shaker for 48 hat 28 uC. Amino acids and isomers in cell-wall hydrolysateswere analysed as described by Hamada et al. (2012). Thecellular fatty acids of strain KIS82-1T consisted of anteiso-C15 : 0 (49.0 %), iso-C16 : 0 (29.1 %), iso-C14 : 0 (12.6 %), iso-C15 : 0 (3.1 %), anteiso-C17 : 0 (3.0 %) and C16 : 0 (1.9 %).Strain KIS82-1T contained diphosphatidylglycerol, phos-phatidylglycerol, three unknown glycolipids, an amino-phospholipid and two unknown lipids (Fig. S3).Menaquinones of strain KIS82-1T were MK-11 (62 %),MK-10 (28 %) and MK-12 (10 %). The peptidoglycan ofstrain KIS82-1T contained glutamic acid, glycine, alanine,homoserine (Hsr) and 2,4-diaminobutyric acid (DAB) in amolar ratio of 1.0 : 1.2 : 0.8 : 0.5 : 0.7. Enantiomeric analysis

of the peptidoglycan amino acids revealed the presence ofD-Glu, Gly, D-Ala, L-Hsr and L-DAB. These data suggestedthat the cell-wall peptidoglycan of strain KIS82-1T was oftype B1b, with L-DAB as the diamino acid. The cellularfatty acids of strain GRS42T were summed feature 8(C18 : 1v6c and/or C18 : 1v7c; 80.6 %), C14 : 0 2-OH (7.8 %),anteiso-C15 : 0 (3.9 %), C16 : 0 (1.7 %), iso-C16 : 0 (1.6 %),summed feature 1 (iso-C15 : 1 H and/or C13 : 0 3-OH)(1.2 %) and C14 : 0 (0.7 %). Strain GRS42T contained dipho-sphatidylglycerol, phosphatidylglycerol, three unknownglycolipids, an unknown phospholipid and one unknownlipid (Fig. S3). Menaquinones of strain GRS42T were MK-8(63 %), MK-9 (24 %) and MK-7 (13 %). The peptidoglycanof strain GRS42T contained glutamic acid, glycine, alanine,homoserine and ornithine (Orn) in a molar ratio of1.0 : 1.1 : 0.8 : 0.3 : 0.9. Enantiomeric analysis of the peptido-glycan amino acids revealed the presence of D-Glu, Gly, D-Ala, L-Hsr and D-Orn. These data suggested that the cell-wallpeptidoglycan of strain GRS42T was of type B2b, with D-Ornas the diamino acid.

For the measurement of DNA–DNA hybridization andDNA G+C contents, genomic DNA was extracted andpurified by the method described by Ausubel et al. (1987).DNA–DNA hybridization was carried out in triplicate asdescribed by Seldin & Dubnau (1985). Probe labelling wasconducted using the non-radioactive DIG High Prime

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Lysinimonas kribbensis MSL-13T (EF466129)Lysinimonas soli SGM3-12T (JN378395)

Microcella putealis CV-2T (AJ717388)Chryseoglobus frigidaquae CW1T (EF373534)

Labedella gwakjiensis KSW2-17T (DQ533552)Amnibacterium kyonggiense KSL51201-037T (FJ527819)Cryobacterium psychrophilum DSM 4854T (AJ544063)

Klugiella xanthotipulae 44C3T (AY372075)Alpinimonas psychrophila Cr8-25T (GU784868)

Galbitalea soli KIS82-1T (JX876866)Frigoribacterium mesophilum MSL-08T (EF466126)Marisediminicola antarctica ZS314T (GQ496083)

Frigoribacterium faeni 801T (Y18807)Frondihabitans sucicola GRS42T (JX876867)Frondihabitans cladoniiphilus CafT13T (FN666417)

Frondihabitans peucedani RS-15T (FM998017)Frondihabitans australicus E1HC-02T (DQ525859)

Compostimonas suwonensis SMC46T (JN000316)

Clavibacter michiganensis DSM 46364T (X77435)Rathayibacter rathayi DSM 748T (X77439)Mycetocola saprophilus CM-01T (AB012647)

Curtobacterium citreum DSM 20528T (X77436)Pseudoclavibacter helvolus DSM 20419T (X77440)

Gulosibacter molinativorax ON4T (AJ306835)Brevibacillus brevis NBRC 15304T (AB271756)

Fig. 1. Neighbour-joining phylogenetic tree based on 16S rRNA gene sequences, showing the taxonomic positions of strainsKIS82-1T and GRS42T in the family Microbacteriaceae. Filled circles indicate that the corresponding nodes were alsorecovered in trees generated with the maximum-parsimony and maximum-likelihood algorithms. Bootstrap values .70 % basedon 1000 replications are shown at branching points. Bar, 0.02 substitutions per nucleotide position.

S.-J. Kim and others

574 International Journal of Systematic and Evolutionary Microbiology 64

DNA Labelling and Detection Starter kit II (RocheMolecular Biochemicals). Hybridized DNA was visualizedusing the DIG Luminescent Detection kit (RocheMolecular Biochemicals). DNA–DNA relatedness wasquantified with a densitometer (Bio-Rad). The G+Ccontent was determined by the fluorometric method(Gonzalez & Saiz-Jimenez, 2002) using SYBR green 1and a real-time PCR thermocycler (Bio-Rad). DNA–DNAhybridization between strain KIS82-1T and Frondihabitanspeucedani KACC 14983T and Frigoribacterium mesophilumKACC 20989T gave relatedness values of 22±4 %(reciprocal value, 28±5 %) and 28±3 %, respectively.DNA–DNA hybridization of strain GRS42T withFrondihabitans peucedani KACC 14983T, Frondihabitanscladoniiphilus KACC 15540T, Frondihabitans australicusKACC 20985T and Frigoribacterium faeni KACC 20509T

resulted in hybridization values of 42±2 % (reciprocal value,49±3 %), 46±5, 33±2 and 30±3 %, respectively. Thegenomic DNA G+C contents of strains KIS82-1T andGRS42T were 63.4 and 65.4 mol%, respectively.

Based on phylogenetic analyses of the 16S rRNA gene, strainKIS82-1T could not be clearly assigned to any known genuswithin the family Microbacteriaceae, although it was relatedto the genera Frondihabitans and Frigoribacterium. StrainKIS82-1T could be differentiated clearly from members ofthe genera Frigoribacterium and Frondihabitans by chemo-taxonomic data such as the peptidoglycan type, menaqui-none composition and fatty acid profile. The cell wall ofstrain KIS82-1T consisted of type B1b peptidoglycan with L-DAB as the diamino acid, while the cell walls of members ofthe genera Frigoribacterium and Frondihabitans are of typeB2b with lysine and ornithine, respectively, as the diaminoacids. Strain KIS82-1T contained MK-11, MK-10 and MK-12 as the major menaquinones; in contrast, members of thegenera Frigoribacterium and Frondihabitans have MK-9 anda combination of MK-7, MK-8 and MK-9, respectively, aspredominant menaquinones. The major fatty acids ofKIS82-1T were anteiso-C15 : 0, iso-C16 : 0 and iso-C14 : 0,compared with anteiso-C15 : 0, iso-C16 : 0, C18 : 1v7c andanteiso-C17 : 0 for the genus Frigoribacterium and summedfeature 8 (C18 : 1v6c and/or C18 : 1v7c), C14 : 0 2-OH andanteiso-C15 : 0 for the genus Frondihabitans (Table 1). Table1 provides differentiating characteristics for strain KIS82-1T

and members of closely related genera within the familyMicrobacteriaceae. According to the phylogenetic trees (Figs1 and S1), strain GRS42T was shown to be a member of thegenus Frondihabitans. The low DNA–DNA relatedness (lessthan 50 %) between strain GRS42T and type strains ofclosely related species of the genus Frondihabitans suggestthat this strain represents a novel genomic species withinthe genus Frondihabitans (Wayne et al., 1987).Furthermore, phenotypic properties such as motility,growth temperature range, substrate assimilation pattern,enzymic activities and quantitative and qualitative differ-ences in the fatty acid composition support the proposal ofGRS42T as the type strain of a novel species within the genusFrondihabitans (Tables 2 and 3). Ta

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Description of Galbitalea gen. nov.

Galbitalea (Gal.bi.ta9le.a. L. adj. galbus yellow; L. fem. n.talea rod; N.L. fem. n. Galbitalea a yellow rod).

Cells are aerobic, Gram-stain-positive, mesophilic, rod-shaped and motile. Catalase- and oxidase-negative. Thecell-wall peptidoglycan is of type B1b with L-DAB as thediamino acid. Respiratory quinones are MK-11, MK-10and MK-12. Predominant cellular fatty acids (.10 % of thetotal fatty acids) are anteiso-C15 : 0, iso-C16 : 0 and iso-C14 : 0.The major polar lipids are diphosphatidylglycerol, phos-phatidylglycerol and unknown glycolipids. Phylogene-tically, the genus is a member of the family Microbac-teriaceae of the phylum Actinobacteria. The type species isGalbitalea soli.

Description of Galbitalea soli sp. nov.

Galbitalea soli (so9li. L. gen. n. soli of soil).

In addition to the description of the genus, the followingproperties are displayed. Cells are 0.4–0.45 mm wide and1.4–2.3 mm long. Grows at 10–37 uC (optimum, 28–30 uC), pH 4.0–9.0 (optimum pH 7.0) and 0–1.0 % (w/v)NaCl (optimum, 1.0 %). Colonies are yellow. Grows on NAand TSA, but not on MacConkey agar or ISP medium 2.Nitrate is not reduced. Hydrolyses starch, but not casein,cellulose, chitin, DNA, hypoxanthine, Tween 80, L-tyrosineor xanthine. Polar lipids include three unknown glycoli-pids, an aminophospholipid and two unknown lipids.

The type strain, KIS82-1T (5KACC 15520T5NBRC108727T), was isolated from a soil on Kaui island, Taean

Table 2. Phenotypic comparison between strain GRS42T and type strains of other species of the genus Frondihabitans

Strains: 1, GRS42T; 2, Frondihabitans australicus KACC 20985T (Zhang et al., 2007; Greene et al., 2009); 3, Frondihabitans cladoniiphilus KACC

15540T (Cardinale et al., 2011); 4, Frondihabitans peucedani KACC 14983T (Lee, 2010). Data for the reference strains were taken from the references

listed unless indicated otherwise. All strains were positive for aesculin hydrolysis and activities of b-galactosidase, esterase (C4), esterase lipase (C8),

acid phosphatase, naphthol-AS-BI-phosphohydrolase, a-glucosidase, b-glucosidase and a-mannosidase, but negative for nitrate reduction, indole

production, glucose fermentation, arginine dihydrolase, gelatin hydrolysis, lipase (C14), cystine arylamidase, trypsin, a-chymotrypsin and b-

glucuronidase. All strains assimilated D-glucose, N-acetylglucosamine, maltose, inositol, sucrose and melibiose, but not capric acid, adipic acid,

malic acid, trisodium citrate, phenylacetic acid, D-ribose, itaconic acid, suberic acid, sodium malonate, sodium acetate, lactic acid, L-alanine,

potassium 5-ketogluconate, L-serine, L-fucose, propionic acid, valeric acid, L-histidine, potassium 2-ketogluconate, 3-hydroxybutyric acid, 4-

hydroxybenzoic acid or L-proline.

Characteristic 1 2 3 4

Isolation source Sap of Acer mono Leaf litter Thallus of lichen Rhizosphere soil

Colony colour Light yellow White Yellow Yellow

Motility + 2 2 2

Growth temperature (uC) 5–30 15–37 1–30 4–37

Respiratory menaquinones 8 (63 %), 9 (24 %), 7 (13 %) 8 (76 %), 7 (24 %) 8 (67 %), 7 (33 %) 8 (75 %), 9 (15 %), 7 (10 %)

Polar lipids* DPG, PG, 3GL, PL, L DPG, PG, GL, 3PL DPG, PG, 3GL, 3PL, AL PG, 5GL, 3PL, 2L

UreaseD 2 + 2 2

Assimilation of:D

L-Arabinose 2 2 + +

D-Mannose + + 2 +

D-Mannitol + + 2 +

Potassium gluconate 2 + 2 +

L-Rhamnose + + 2 2

Glycogen 2 2 2 +

3-Hydroxybenzoic acid 2 2 + 2

Salicin + + 2 +

D-Sorbitol 2 2 2 +

Enzymic activitiesD

Alkaline phosphatase + 2 2 +

Leucine arylamidase 2 + + +

Valine arylamidase 2 + 2 2

a-Galactosidase 2 + 2 +

N-Acetyl-b-glucosaminidase 2 + 2 2

a-Fucosidase 2 + 2 2

*DPG, Diphosphatidylglycerol; PG, phosphatidylglycerol; AL, unknown aminolipid; GL, unknown glycolipid; PL, unknown phospholipid; L,

unknown lipid.

DData from this study.



county, Republic of Korea. The DNA G+C content of thetype strain is 63.4 mol%.

Description of Frondihabitans sucicola sp. nov.

Frondihabitans sucicola [su.ci9co.la. L. n. sucus sap; L. suff.-cola (from L. n. incola) inhabitant, dweller; N.L. n. sucicolaa sap-dweller].

Cells are aerobic, Gram-stain-positive, mesophilic, rod-shaped and motile. Cells are 0.5–0.7 mm wide and 1.1–1.6 mm long. Colonies are light yellow. Grows at 4–30 uC(optimum, 28 uC), pH 4.0–9.0 (optimum, pH 7.0) and 0–2.0 % (w/v) NaCl (optimum, 1.0 %). Grows on NA, TSA,MacConkey agar and ISP medium 2. Catalase-positive andoxidase-negative. Nitrate is not reduced. Does not hydrolysecasein, cellulose, chitin, DNA, hypoxanthine, starch, Tween80, L-tyrosine or xanthine. Physiological characteristics aregiven in Table 2. Polar lipids are diphosphatidylglycerol,phosphatidylglycerol, three unknown glycolipids, an unknownphospholipid and one unknown lipid. The major cellular fattyacid (.10 % of the total fatty acids) is summed feature 8(C18 : 1v6c and/or C18 : 1v7c). Menaquinones are MK-8, MK-9and MK-7. The peptidoglycan contains glutamic acid, glycine,alanine, homoserine and ornithine in a molar ratio of1.0 : 1.1 : 0.8 : 0.3 : 0.9, and is of type B2b with D-Orn as thediamino acid.

The type strain, GRS42T (5KACC 15521T5NBRC108728T), was isolated from sap of Acer mono growing

on Baikam Mountain, Hwasun county, Republic of Korea.The DNA G+C content of the type strain is 65.4 mol%.

Acknowledgements

This study was carried out with the support of the National Academy

of Agricultural Science, Rural Development Administration, Republic

of Korea (project no. PJ008666). The authors thank Professor J. P.

Euzeby of the Ecole Nationale Veterinaire in Toulouse for advice

concerning the naming of the novel taxa.

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description of galbitalea soli gen. nov., sp. nov., and frondihabitans sucicola sp. nov

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