Journal of Research 5(1):1­13, 2016

Report on 31 unrecorded bacterial species in Korea that belong to the phylum

Jung­Hye Choi1, Ju­Hee Cha1, Jin­Woo Bae2, Jang­Cheon Cho3, Jongsik Chun4, Wan­Taek Im5, Kwang Yeop Jahng6, Che Ok Jeon7, Kiseong Joh8, Seung Bum Kim9, Chi Nam Seong10, Jung­Hoon Yoon11 and Chang­Jun Cha1,*

1Department of Systems Biotechnology, Chung-Ang University, Anseong 17546, Korea 2Department of Biology, Kyung Hee University, Seoul 02447, Korea 3Department of Biological Sciences, Inha University, Incheon 22212, Korea 4School of Biological Sciences, Seoul National University, Seoul 08826, Korea 5Department of Biotechnology, Hankyong National University, Anseong 17579, Korea 6Department of Life Sciences, Chonbuk National University, Jeonju-si 54896, Korea 7Department of Life Science, Chung-Ang University, Seoul 06974, Korea 8Department of Bioscience and Biotechnology, Hankuk University of Foreign Studies, Gyeonggi 17035, Korea 9Department of Microbiology, Chungnam National University, Daejeon 34134, Korea 10Department of Biology, Sunchon National University, Suncheon 57922, Korea 11Department of Food Science and Biotechnology, Sungkyunkwan University, Suwon 16419, Korea

*Correspondent: [email protected]

To discover and characterize indigenous species in Korea, a total of 31 bacterial strains that belong to the phylum Actinobacteria were isolated from various niches in Korea. Each strain showed the high sequence similarity (>99.1%) with the closest bacterial species, forming a robust phylogenetic clade. These strains have not been previously recorded in Korea. According to the recently updated of the phylum Actinobacteria based upon 16S rRNA trees, we report 25 genera of 13 families within 5 orders of the class Actinobacteria as actinobacterial species found in Korea. Cellular morphology, Gram staining, basic biochemical characteristics are described in the species description. Keywords: 16S rRNA gene, Actinobacteria, bacterial diversity, unrecorded species

Ⓒ 2016 National Institute of Biological Resources DOI:10.12651/JSR.2016.5.1.001

although most of prokaryotic species have yet to be un­ IntroductIon cultured. An increasing attention has been paid on the investi­ Microorganisms are the most diverse and abundant gation of new or unrecorded prokaryotic species that are groups of organisms on Earth, and they play important indigenous in Korea. A variety of novel bacterial spe­ roles in various biogeochemical processes (Whitman et cies and unrecorded bacterial species were isolated from al., 1998; Fierer and Jackson, 2006). However, it was various environmental samples collected in Korea. As a proposed that less than only 1% of total microbial cells result, the bacterial isolates were assigned to the phyla were cultured on the planet (Amann et al., 1995). The Actinobacteria, Bacteroidetes, Deinococci, Firmicutes, diversity of uncultured microorganisms can offer the Proteobacteria and Verrucomicrobia. possibility of a vast number of novel microbial taxa to The phylum Actinobacteria is comprised mainly be still discovered. Thanks to innovative cultivation of Gram­positive with a high G+C content methods, increasing numbers of new names of prokar­ (>55 mol% in genomic DNA), and constitutes one of yotic species have been validly published (Oren and the largest phyla within the Bacteria (Gao and Gupta, Garrity, 2014), and approximately 11,500 prokaryotic 2012). This phylum that contains >300 genera (Zhi et species have been validly published so far (Parte, 2014), al., 2009) display enormous diversity in terms of their 2 JOURNAL OF SPECIES RESEARCH Vol. 5, No. 1 morphology, physiology, and metabolic capabilities. The dilution plating technique on various culture media in­ morphologies of actinobacterial species vary from coc­ cluding R2A (BD), marine agar (MA; BD), nutrient agar coid to branched mycelia. Members of Actinobacteria (NA; BD), 1/10 PCA (BD), and minimal medium (MM; are found in a wide range of environments such as soil, 30 g D­glucose, 1 g yeast extract, 0.5 g K2HPO4, 0.5 g water, deep­sea, arctic ice, chemically contaminated KH2PO4, 2.2 g (NH4)2SO4, 0.2 g MgSO4·7H2O, 0.01 g sites, radioactive environments, gastrointestinal tracts of MnSO4·7H2O, 0.01 g FeSO4·7H2O and 0.01g NaCl in 1 humans and animals, and plants (Gao and Gupta, 2012). L of water). The agar plates were incubated at 25-37℃ Recently, the taxonomy of the phylum Actinobacteria for 2­10 days (Table 1). All isolates were sub­cultured, based upon 16S rRNA trees was updated (Ludwig and purified and stored as a stock culture in the same medi­ Klenk, 2005.), which is the basis of the section on Acti- um supplemented with 10­30% (v/v) glycerol. The des­ nobacteria in the Bergey’s Manual of Systematic Bacte­ ignated strain IDs, sources, culture media, and incuba­ riology. Previously, the phylum Actinobacteria consisted tion conditions are presented in Table 1. of one class, 5 subclasses, and 9 orders. In the revised Genomic DNA extraction, PCR amplification and 16S taxonomy, subclasses and suborders were eliminated, rRNA gene sequencing were carried out using standard and elevated to the ranks of classes and orders, respec­ procedures; the 16S rRNA gene of the isolates was am­ tively; the phylum Actinobacteria is now divided into 6 plified using the universal bacterial primer pair (27F and classes and 22 orders. The largest class Actinobacteria, 1492R). The 16S rRNA gene sequences were compared which accounts for >80% of all known actinobacte­ with sequences in the ExTaxon­e server (http://eztaxon­ rial families/genera, now contains a total of 15 orders, e.ezbiocloud.net/) (Kim et al., 2012) and aligned with including both previously proposed orders Actinomy- closely related species by using the CLUSTAL X program cetales and Bifidobacteriales (Zhi et al., 2009). How­ (Thompson et al., 1997). The phylogenetic relationships ever, the order Actinomycetales is now restricted to the were evaluated by using neighbor­joining (Saitou and members of the family Actinomycetaceae, and the other Nei, 1987) and maximum­likelihood (Felsen­stein, 1981) suborders that were previously part of this order are now algorithms, and the trees were constructed using the Mega designated as distinct orders. 6 program (Tamura et al., 2013). Evolutionary distances The species of this group also display tremendous were calculated by the model of Jukes and Cantor (1969) physiological diversity, as revealed by their production and tree topologies were evaluated based on bootstrap of numerous extracellular enzymes (Chater et al., 2010) analyses of 1,000 data sets. and secondary metabolites, many of which are anti­ Colony morphology was observed on agar plates after biotics (Hopwood, 2007). The most extensively stud­ cells grew up to stationary phase, and cellular morphol­ ied representatives of this group include soil­dwelling ogy and cell size were determined by either transmission Streptomyces spp., which are the major producers of an­ electron microscopy or scanning electron microscopy tibiotics (Chater, 2006; Ventura et al., 2007); the genus using cells grown on various agar plates for 2­10 days. Mycobacterium, which are important human pathogens The Gram staining was carried out using the Gram­stain­ and responsible for the largest number of human deaths ing kit (Sigma­Aldrich), and enzyme activities and uti­ from bacterial infections; and the genus Rhodococcus, lization of different carbon sources were assessed using which possesses a high potential for industrial and envi­ API 20NE galleries (bioMérieux) according to the man­ ronmental applications (Martínková et al., 2009; Yam et ufacturer’s instructions. al., 2011). As a part of results obtained from the research pro­ gram supported by NIBR, the present report focuses on results and dIscussIon the description of bacterial species belonging to the Ac- Assignment of unrecorded strains to the validly tinobacteria which have not been previously isolated in published species of Actinobacteria Korea. Here, we report 31 unrecorded bacterial species that belong to the phylum Actinobacteria. On the basis of 16S rRNA gene sequence compar­ isons and phylogenetic analyses, a total of 31 strains, which have not been previously isolated in Korea, were MaterIals and Methods assigned to the phylum Actinobacteria. All of the 31 strains belonged to the class Actinobacteria and were A total of 31 bacterial strains were isolated from vari­ distributed to 13 families of 5 orders; 8 strains of 7 gen­ ous environmental samples collected from soil, tidal flat, era in the family , 4 strains of the ge­ freshwater, seawater, wetland, and plant roots using var­ nus Rhodococcus in the family Nocardiaceae, 4 strains ious culture media (Table 1). The strains were isolated of 3 genera in the family Micrococcaceae, 3 strains of 3 from each environmental sample by using the standard genera in the family , 3 strains of the February 2016 CHOI ET AL.­UNRECORDED ACTINOBACTERIA SPECIES 3 Incubation conditions 25℃, 2d 30℃, 2d 25℃, 2d 25℃, 10d 30℃, 2d 30℃, 2d 25℃, 3d 25℃, 2d 30℃, 2d 25℃, 5d 25℃, 3d 25℃, 2d 30℃, 2d 25℃, 2d 30℃, 2d 25℃, 2d 25℃, 2d 25℃, 3d 25℃, 3d 37℃, 2d 30℃, 2d 25℃, 3d Medium MA R2A R2A 1/10 PCA MA R2A MM R2A MA MM R2A MA R2A R2A MA MA MA MA R2A R2A R2A MM Isolation source Sea water Plant root Wetland Fresh water Soil Soil Soil Wetland flat Tidal Soil Sea water flat Tidal Plant root Fresh water Soil Soil Soil Soil Plant Fresh water Soil Soil 99.9 99.4 99.9 99.2 99.7 99.3 99.8 99.6 99.6 99.3 99.9 100 99.4 99.9 99.3 99.3 99.9 99.7 99.4 99.4 99.7 99.1 Similarity (%) Most closely related species Dietzia cinnamea Mycobacterium frederiksbergense Rhodococcus equi Rhodococcus canchipurensis Rhodococcus jostii Rhodococcus maanshanensis Tsukamurella tyrosinosolvens Cellulomonas xylanilytica Brachybacterium muris Humibacillus xanthopallidus Serinicoccus profundi Janibacter anophelis flavus Curtobacterium citreum esteraromaticum Leifsonia soli Microbacterium pumilum Microbacterium oxydans baldri Agrococcus Aquiluna rubra Arthrobacter globiformis Arthrobacter scleromae NIBR ID NIBRBA0000114113 NIBRBA0000113880 NIBRBA0000113883 NIBRBA0000113918 NIBRBA0000113950 NIBRBA0000113951 NIBRBA0000113963 NIBRBA0000113882 NIBRBA0000114094 NIBRBA0000113978 NIBRBA0000113995 NIBRBA0000114044 NIBRBA0000113879 NIBRBA0000113848 NIBRBA0000113944 NIBRBA0000113962 NIBRBA0000113972 NIBRBA0000113997 NIBRBA0000114009 NIBRBA0000114076 NIBRBA0000113945 NIBRBA0000113977 ­ RY7 Strain ID KYW853 SPE2 ­ 2 UKS ­ 28 IK56 MK5 ­ 14 RS5 ­ 4 mNW17 UKS ­ 33 HME8794 mNW13 HWR24 AB7 SPE ­ 06 MIC10 MS5 ­ 22 MA9 MAT14 WR ­ M1Y WT HME8543 5 ­ 1 RK 4Y mNW18 and their taxonomic assignment based on 16S rRNA gene analysis. and their taxonomic assignment based on 16S rRNA Genus Dietzia Mycobacterium Rhodococcus Rhodococcus Rhodococcus Rhodococcus Tsukamurella Cellulomonas Brachybacterium Humibacillus Serinicoccus Janibacter Plantibacter Curtobacterium Microbacterium Leifsonia Microbacterium Microbacterium Agrococcus Aquiluna Arthrobacter Arthrobacter Actinobacteria Family Dietziaceae Mycobacteriaceae Nocardiaceae Tsukamurellaceae Cellulomonadaceae Dermabacteraceae Intrasporangiaceae Microbacteriaceae Micrococcaceae Order Corynebacteriales List of strains belonging to the phylum Table 1. Table Class Actinobacteria 4 JOURNAL OF SPECIES RESEARCH Vol. 5, No. 1

genus Streptomyces in the family Streptomycetaceae, 2 strains of 2 genera in the family Promicromonospora- ceae, and 7 strains of 7 genera in the respective families Incubation conditions 25℃, 3d 30℃, 2d 30℃, 2d 30℃, 2d 25℃, 2d 25℃, 2d 30℃, 2d 25℃, 2d 25℃, 2d Dietziaceae, Mycobacteriaceae, Tsukamurellaceae, Cel- lulomonadaceae, Dermabacteraceae, Propionibacteria- R2A MA R2A NA R2A R2A MA MA R2A ceae and Streptosporangiaceae (Table 1). These strains Medium were Gram­staining­positive, chemoheterotrophic, and rod­shaped, except for 6 strains that were Gram­stain­ ing­negative and 5 strains that were coccoid­shaped (Fig.

Isolation source flat Tidal flat Tidal Soil Soil Soil Wetland Soil Soil Wetland 1). Colony size, morphology, and physiological charac­ teristics are also shown in the species description. Comparative sequence analyses of these strains re­

99.6 99.1 100 99.9 99.2 100 100 100 99.6 vealed high 16S rRNA gene sequence similarities (> Similarity (%) 99.1%) with the closest relatives (Table 1). The phyloge­ netic analyses showed that each strain formed a robust phylogenetic clade with the most closely related species. Eight strains belonging to the family Microbacteriaceae were phylogenetically assigned to . rosea (Behrendt et al., 2002), Curtobacterium citreum (Yamada Most closely related species Kocuria rosea Micrococcus flavus Isoptericola nanjingensis Cellulosimicrobium funkei soli Microlunatus Streptomyces avellaneus Streptomyces omiyaensis Streptomyces atratus rosea Microbispora subsp and Komagata, 1972), Microbacterium esteraromaticum (Takeuchi and Hatano, 1998), Leifsonia soli (Madhai­ yan et al., 2010), Microbacterium pumilum (Kageyama et al., 2006), Microbacterium oxydans (Schumann et al., 1999), Agrococcus baldri (Zlamala et al., 2002) and Aquiluna rubra (Hahn, 2009), respectively. Four strains of the family Micrococcaceae were assigned to Arthro-

NIBR ID NIBRBA0000113993 NIBRBA0000114092 NIBRBA0000113943 NIBRBA0000113954 NIBRBA0000113889 NIBRBA0000113885 NIBRBA0000113953 NIBRBA0000113970 NIBRBA0000113886 bacter globiformis (Conn and Dimmick, 1947), Arthro- bacter scleromae (Ying Huang et al., 2005), Kocuria rosea (Stackebrandt et al., 1995) and Micrococcus flavus (Liu et al., 2007), respectively. Four strains were as­ Strain ID HR39 HME8781 RS5 ­ 5_B NS3 ­ 4_B Gsoil 950 UKS ­ 24 MS5 ­ 13 MA10 UKS ­ 23 signed to the genus Rhodococcus in the family Nocardi- aceae; R. equi (Goodfellow and Alderson, 1977), R. can- chipurensis (Nimaichand et al., 2013), R. jostii (Takeuchi et al., 2002) and R. maanshanensis (Zhang et al., 2002). Eight strains were assigned to Humibacillus xanthopalli- dus (Kageyama et al., 2008), Serinicoccus profundi (Xiao Genus Kocuria Micrococcus Isoptericola Cellulosimicrobium Microlunatus Streptomyces Streptomyces Streptomyces Microbispora et al., 2011) and Janibacter anophelis (Peter Kämpfer et al., 2006) of the family Intrasporangiaceae, Isoptericola

nanjingensis (Zhi Huang et al., 2012) and Cellulosimi- crobium funkei (Brown et al., 2006) of the family Promi- cromonosporaceae, and Streptomyces avellaneus (Bal­ dacci and Grein, 1966), S. omiyaensis (Umezawa et al., 1950) and S. atratus (Shibata et al., 1962) of the family Family Micrococcaceae Promicromono- sporaceae Propionibacteriaceae Streptomycetaceae Streptosporangiaceae Streptomycetaceae, respectively. Seven strains were as­ signed to Dietzia cinnamea (Yassin et al., 2006) of the family Dietziaceae, Mycobacterium frederiksbergense (Willumsen et al., 2001) of the family Mycobacteriace- ae, Tsukamurella tyrosinosolvens (Yassin et al., 1997) of the family Tsukamurellaceae, Cellulomonas xylanilytica Order Micrococcales Propionibacteriales Streptomycetales Streptosporangiales (Rivas et al., 2004) of the family Cellulomonadaceae, Brachybacterium muris (Buczolits et al., 2003) of the

Continued. family Dermabacteraceae, Microlunatus soli (P. Kämp­ fer et al., 2010) of the family Propionibacteriaceae, and

Table 1. Table Class Actinobacteria Microbispora rosea subsp. rosea (Miyadoh et al., 1990) February 2016 CHOI ET AL.­UNRECORDED ACTINOBACTERIA SPECIES 5 of the family Streptosporangiaceae, respectively (Fig. 2) rod­shaped. Colonies are smooth, circular, convex, opa­ que and pallid pink­colored after 10 days of incubation Description of Dietzia cinnamea KYW853 on 1/10 PCA at 25°C. Positive reactions are obtained for nitrate reduction, esculin hydrolysis and β-galactosidase Cells are Gram-staining-negative, non-flagellated and in API 20NE, and negative reactions are obtained for in­ rod­shaped. Colonies are opaque, round, smooth, con­ dole production, glucose fermentation, arginine dihydro­ vex and orange­colored after 2 days of incubation on lase, urease and gelatinase. Malic acid is utilized. Does MA at 25℃. Negative reactions are obtained for nitrate not utilize D­glucose, L­arabinose, D­mannose, D­man­ reduction, indole production, glucose fermentation, nitol, N­acetyl­glucosamine, D­maltose, potassium glu­ arginine dihydrolase, urease, esculin hydrolysis, gelati­ conate, capric acid, adipic acid, trisodium citrate and nase and β-galactosidase in API 20NE. Does not utilize phenylacetic acid. Strain IK56 (=NIBRBA0000113918) D­glucose, L­arabinose, D­mannose, D­mannitol, N­ was isolated from a freshwater sample, Ingyeong lake, acetyl­glucosamine, D­maltose, potassium gluconate, Incheon, Korea. capric acid, adipic acid, malic acid, trisodium citrate = and phenylacetic acid. Strain KYW853 ( NIBRBA Description of Rhodococcus jostii MK5-14 0000114113) was isolated from a sea water sample, Gwangyang Bay, Gwangyang, Korea. Cells are Gram-staining-positive, non-flagellated and rod­shaped. Colonies are circular, entire, smooth and Description of Mycobacterium frederiksbergense beige colored after 2 days on MA at 30°C. Negative re­ SPE2-2 actions are obtained for nitrate reduction, indole produc­ tion, glucose fermentation, arginine dihydrolase, urease, Cells are Gram-staining-positive, non-flagellated and esculin hydrolysis, gelatinase and β-galactosidase in API rod­shaped. Colonies are round, smooth and yellow­col­ 20NE. D­glucose, L­arabinose, D­mannose, D­manni­ ored after 2 days of incubation on R2A at 30℃. Positive tol, N­acetyl­glucosamine, potassium gluconate, adipic reactions are obtained for nitrate reduction, glucose fer­ acid, malic acid, trisodium citrate and phenylacetic acid mentation, esculin hydrolysis and β-galactosidase in API are utilized. Does not utilize D­maltose and capric acid. 20NE, and negative reactions are obtained for indole Strain MK5­14 ( =NIBRBA0000113950) was isolated production, arginine dihydrolase, urease and gelatinase. from a sample of ginseng field, Iljuk, Anseong, Korea. D­glucose, L­arabinose, D­mannose, D­mannitol, N­ace­ tyl­glucosamine, potassium gluconate, adipic acid, malic acid, trisodium citrate and phenylacetic acid are utilized. Description of Rhodococcus maanshanensis RS5-4 Does not utilize D­maltose and capric acid. Strain SPE2­ Cells are Gram-staining-positive, non-flagellated, non-

2 ( =NIBRBA0000113880) was isolated from a plant pigmented and rod­shaped. Colonies are circular, entire, root sample, Chungnam National University, Daejeon, smooth and white­colored after 2 days on R2A at 30°C. Korea. Positive reactions are obtained for nitrate reduction, urease, esculin hydrolysis, and β-galactosidase in API Description of Rhodococcus equi UKS-28 20NE and negative reactions are obtained for indole Cells are Gram-staining-positive, non-flagellated and production, glucose fermentation, arginine dihydrolase rod­shaped. Colonies are circular, raised, entire and yel­ and gelatinase. D­glucose, L­arabinose, D­mannose, low­colored after 2 days of incubation on R2A at 25°C. D­mannitol, N­acetyl­glucosamine, D­maltose, potassi­ Positive reaction is obtained for nitrate reduction in API um gluconate, adipic acid, malic acid and trisodium ci­ 20NE, and negative reactions are obtained for indole trate are utilized. Does not utilize capric acid and pheny­ = production, glucose fermentation, arginine dihydrolase, lacetic acid. Strain RS5­4 ( NIBRBA0000113951) was urease, esculin hydrolysis, gelatinase and β-galactosi­ isolated from a sample of ginseng field, Iljuk, Anseong, dase. Malic acid is utilized. Does not utilize D­glucose, Korea. L­arabinose, D­mannose, D­mannitol, N­acetyl­glucos­ amine, D­maltose, potassium gluconate, capric acid, Description of Tsukamurella tyrosinosolvens mNW17 adipic acid, trisodium citrate and phenylacetic acid. Cells are Gram-staining-positive, non-flagellated and Strain UKS­28 ( =NIBRBA0000113883) was isolated rod­shaped. Colonies are circular and beige­colored after from a wetland sample, Ungok­ri, Gochang, Chonbuk, 3 days on minimal medium at 25℃. Positive reactions Korea. are obtained for glucose fermentation, urease, esculin hydrolysis, gelatinase and β-galactosidase in API 20NE, Description of Rhodococcus canchipurensis IK56 and negative reactions are obtained for nitrate reduction, Cells are Gram-staining-positive, non-flagellated and indole production and arginine dihydrolase. L­arabinose, 6 JOURNAL OF SPECIES RESEARCH Vol. 5, No. 1

Fig. 1. Transmission electron micrographs or scanning electron micrographs of cells of the strains isolated in this study. Strains: 1, KYW853; 2, SPE2­2; 3, UKS­28; 4, IK56; 5, MK5­14; 6, RS5­4; 7, mNW17; 8, UKS­33; 9, HME8794; 10, mNW13; 11, HWR24; 12, AB7; 13, SPE­06; 14, MIC10; 15, MS5­22; 16, MA9; 17, MAT14; 18, WR­M1Y; 19, WT­RY7; 20, HME8543; 21, RK 4Y 5­1; 22, mNW18; 23, HR39; 24, HME8781; 25, RS5­5_B; 26, NS3­4_B; 27, Gsoil 950; 28, UKS­24; 29, MS5­13; 30, MA10; 31, UKS­23. February 2016 CHOI ET AL.­UNRECORDED ACTINOBACTERIA SPECIES 7

Fig. 2. Neighbor­joining phylogenetic tree based on 16S rRNA gene sequences, showing the relationship between the strains isolated in this study and their relatives of the phylum Actinobacteria. The numbers at nodes represent bootstrap values (>50%) obtained by neigh­ bor­joining and maximum­likelihood methods, respectively. Closed circles indicate the nodes recovered by maximum­likelihood algorithm. The GenBank accession number of each species is enclosed in parentheses. Bar, 0.02 substitutions per nucleotide position. 8 JOURNAL OF SPECIES RESEARCH Vol. 5, No. 1

D­mannitol, N­acetyl­glucosamine, D­maltose, potassi­ Description of Serinicoccus profundi HWR24 um gluconate, trisodium citrate and phenylacetic acid are Cells are Gram-staining-positive, non-flagellated and utilized. Does not utilize D­glucose, D­mannose, capric coccoid­shaped. Colonies are circular, convex, glistening acid, adipic acid and malic acid. Strain mNW17 ( =NI and yellow­colored after 3 days on R2A at 25°C. Posi­ BRBA0000113963) was isolated from a forest soil sam­ tive reactions are obtained for nitrate reduction, esculin ple, Gwanak Mountain, Seoul, Korea. hydrolysis, gelatinase and β-galactosidase in API 20NE, and negative reactions are obtained for indole pro­ Description of Cellulomonas xylanilytica UKS-33 duction, glucose fermentation, arginine dihydrolase and Cells are Gram-staining-positive, non-flagellated and urease. D­glucose, D­mannose, D­maltose, potassium rod­shaped. Colonies are circular, raised, entire and yel­ gluconate and malic acid are utilized. Does not utilize low­colored after 2 days on R2A at 25°C. Positive reac­ L­arabinose, D­mannitol, N­acetyl­glucosamine, capric tions are obtained for esculin hydrolysis and β-galacto­ acid, adipic acid, trisodium citrate and phenylacetic acid. sidase in API 20NE, and negative reactions are obtained Strain HWR24 ( =NIBRBA0000113995) was isolated for nitrate reduction, indole production, glucose fermen­ from a tidal flat sample, Hwango-ri, Anmyeon-eup, Tae­ tation, arginine dihydrolase, urease and gelatinase. Does an, Korea. not utilize D­glucose, L­arabinose, D­mannose, D­man­ nitol, N­acetyl­glucosamine, D­maltose, potassium glu­ Description of Janibacter anopheles AB7 conate, capric acid, adipic acid, malic acid, trisodium Cells are Gram-staining-positive, non-flagellated and citrate and phenylacetic acid. Strain UKS­33 ( =NIBR short­rod­shaped. Colonies are circular, raised, entire BA0000113882) was isolated from a wetland sample, and ivory­colored after 2 days on MA at 25°C. Positive Ungok­ri, Gochang, Korea. reactions are obtained for nitrate reduction, esculin hy­ drolysis and β-galactosidase in API 20NE and negative Description of Brachybacterium muris HME8794 reactions are obtained for indole production, glucose fer­ Cells are Gram-staining-positive, non-flagellated and mentation, arginine dihydrolase, urease and gelatinase. rod­shaped. Colonies are circular, convex, entire and D­glucose, L­arabinose, D­mannose, D­mannitol, N­ace­ yellow-colored after 2 days on MA at 30℃. Positive tyl­glucosamine, D­maltose, potassium gluconate, malic reactions are obtained for esculin hydrolysis and β-ga­ acid and trisodium citrate are utilized. Does not utilize lactosidase in API 20NE, and negative reactions are ob­ capric acid, adipic acid and phenylacetic acid. Strain tained for nitrate reduction, indole production, glucose AB7 (=NIBRBA0000114044) was isolated from a tidal fermentation, arginine dihydrolase, urease and gelati­ flat sample, Uihang-ri, Sowon-myeon, Taean, Korea. nase. D­glucose, D­mannose, D­maltose and potassium gluconate are utilized. Does not utilize L­arabinose, D­ Description of Plantibacter flavus SPE-06 mannitol, N­acetyl­glucosamine, capric acid, adipic acid, Cells are Gram-staining-positive, non-flagellated and malic acid, trisodium citrate and phenylacetic acid. Strain rod and spiral­shaped. Colonies are round, convex and HME8794 ( =NIBRBA0000114003) was isolated from yellow­colored after 2 days on R2A at 30°C. Positive a tidal flat sample, Sinan, Korea. reactions are obtained for glucose fermentation, esculin hydrolysis and β-galactosidase in API 20NE, and nega­ Description of Humibacillus xanthopallidu mNW13 tive reactions are obtained for nitrate reduction, indole Cells are flagellated and rod-shaped. Colonies are cir- production, arginine dihydrolase, urease and gelatinase. cular and orange­colored after 5 days on minimal medi­ D­glucose, L­arabinose, D­mannose, D­mannitol, N­ace­ um at 25℃. Positive reactions are obtained for glucose tyl­glucosamine, D­maltose, potassium gluconate and fermentation, urease, esculin hydrolysis and β-galac­ malic acid are utilized. Does not utilize capric acid, adip­ tosidase in API 20NE, and negative reactions are ob­ ic acid, trisodium citrate and phenylacetic acid. Strain tained for nitrate reduction, indole production, arginine SPE­06 ( =NIBRBA0000113879) was isolated from a dihydrolase and gelatinase. D­mannose, D­mannitol, plant root sample, Chungnam National University, Dae­ D­maltose and malic acid are utilized. Does not utilize jeon, Korea. D­glucose, L­arabinose, N­acetyl­glucosamine, potas­ sium gluconate, capric acid, adipic acid, trisodium ci­ Description of Curtobacterium citreum MIC10 trate and phenylacetic acid. Strain mNW13 ( =NIBR Cells are Gram-staining-negative, non-flagellated and BA0000113978) was isolated from a forest soil sample, rod­shaped. Colonies are circular and yellow­colored Gwanak Mountain, Seoul, Korea. after 2 days on R2A at 25°C. Positive reactions are ob­ tained for nitrate reduction, esculin hydrolysis, gelati­ February 2016 CHOI ET AL.­UNRECORDED ACTINOBACTERIA SPECIES 9 nase and β-galactosidase in API 20NE, and negative re­ nylacetic acid. Strain MAT14 (=NIBRBA0000113972) actions are obtained for indole production, glucose fer­ was isolated from a forest soil sample, Gwanak Moun­ mentation, arginine dihydrolase and urease. D­glucose, tain, Seoul, Korea. L­arabinose, D­mannose, D­mannitol, N­acetyl­glucos­ amine, D­maltose, and potassium gluconate are utilized. Description of Microbacterium oxydans WR-M1Y Does not utilize capric acid, adipic acid, malic acid, tri­ Cells are Gram-staining-positive, non-flagellated and sodium citrate and phenylacetic acid. Strain MIC10 rod­shaped. Colonies are circular, convex, glistening and (=NIBRBA0000113848) was isolated from a fresh wa­ orange/yellow-colored after 3 days on MA at 25℃. Pos­ ter, Jeju island, Korea. itive reactions are obtained for esculin hydrolysis, gelati­ nase and β-galactosidase in API 20NE, and negative Description of Microbacterium esteraromaticum reactions are obtained for nitrate reduction, indole pro­ MS5-22 duction, glucose fermentation, arginine dihydrolase and Cells are Gram-staining-negative, flagellated and rod- urease. D­glucose, D­mannose, D­mannitol, D­maltose shaped. Colonies are circular, entire, smooth and yel­ and potassium gluconate are utilized. Does not utilize low-colored after 2 days on MA at 30℃. Positive reac- L­arabinose, N­acetyl­glucosamine, capric acid, adipic tions are obtained for nitrate reduction and β-galactosi­ acid, malic acid, trisodium citrate and phenylacetic acid. = dase in API 20NE, and negative reactions are obtained Strain WR­M1Y ( NIBRBA0000113997) was isolated for indole production, glucose fermentation, arginine from a soil sample in field of reeds, Wando, Korea. dihydrolase, urease, esculin hydrolysis and gelatinase. D­glucose, L­arabinose, D­mannose, D­mannitol, N­ace­ Description of Agrococcus baldri WT-RY7 tyl­glucosamine, D­maltose and potassium gluconate Cells are Gram-staining-positive, flagellated and rod- utilized. Does not utilize capric acid, adipic acid, ma­ shaped. Colonies are circular, convex, glistening and viv­ lic acid, trisodium citrate and phenylacetic acid. Strain id-yellow-colored after 3 days on R2A at 25℃. Positive = MS5­22 ( NIBRBA0000113944) was isolated from a reactions are obtained for esculin hydrolysis, gelatinase sample of ginseng field, Iljuk, Anseong, Korea. and β-galactosidase in API 20NE, and negative reactions are obtained for nitrate reduction, indole production, Description of Leifsonia soli MA9 glucose fermentation, arginine dihydrolase and urease. Does not utilize D­glucose, L­arabinose, D­mannose, Cells are flagellated and rod-shaped. Colonies are cir­ D­mannitol, N­acetyl­glucosamine, D­maltose, potas­ cular and yellow-colored after 2 days on MA at 25℃. sium gluconate, capric acid, adipic acid, malic acid, Positive reactions are obtained for urease, esculin hy­ trisodium citrate and phenylacetic acid. Strain WT­RY7 drolysis and β-galactosidase in API 20NE, and nega­ (=NIBRBA0000114009) was isolated from a plant sam­ tive reactions are obtained for nitrate reduction, indole ple, Wando, Korea. production, glucose fermentation, arginine dihydrolase and gelatinase. D­glucose and D­mannose are utilized. Description of Aquiluna rubra HME8543 Does not utilize L­arabinose, D­mannitol, N­acetyl­glu­ cosamine, D­maltose, potassium gluconate, capric acid, Cells are Gram­staining­positive, non­flagellated adipic acid, malic acid, trisodium citrate and pheny­ and rod­shaped. Colonies are circular, raised, entire lacetic acid. Strain MA9 (=NIBRBA0000113962) was and red-colored after 2 days on R2A at 37℃. Positive isolated from a forest soil sample, Gwanak Mountain, reaction is obtained for β-galactosidase in API 20NE, Seoul, Korea. and negative reactions are obtained for nitrate reduc­ tion, indole production, glucose fermentation, arginine Description of Microbacterium pumilum MAT14 dihydrolase, urease, esculin hydrolysis and gelatinase. Does not utilize D­glucose, L­arabinose, D­mannose, Cells are Gram-staining-positive, flagellated and rod- D­mannitol, N­acetyl­glucosamine, D­maltose, potassi­ shaped. Colonies are circular and orange­colored after um gluconate, capric acid, adipic acid, malic acid, triso­ 2 days on MA at 25℃. Positive reactions are obtained dium citrate and phenylacetic acid. Strain HME8543 (= for glucose fermentation, urease, esculin hydrolysis and NIBRBA0000114076) was isolated from a freshwater β-galactosidase in API 20NE, and negative reactions are sample, Geongan Stream, Yongin, Korea. obtained for nitrate reduction, indole production, argi­ nine dihydrolase and gelatinase. Does not utilize D­glu­ Description of Arthrobacter globiformis RK 4Y 5-1 cose, L­arabinose, D­mannose, D­mannitol, N­acetyl­ glucosamine, D­maltose, potassium gluconate, capric Cells are Gram-staining-negative, flagellated, non-pig­ acid, adipic acid, malic acid, trisodium citrate and phe­ mented and rod­shaped. Colonies are circular, entire, 10 JOURNAL OF SPECIES RESEARCH Vol. 5, No. 1 smooth and white-colored after 2 days on R2A at 30℃. Does not utilize L­arabinose, D­mannose, D­mannitol, Positive reactions are obtained for gelatinase and β-ga­ N­acetyl­glucosamine, potassium gluconate, capric acid, lactosidase in API 20NE, and negative reactions are ob­ adipic acid, trisodium citrate and phenylacetic acid. tained for nitrate reduction, indole production, glucose Strain HME8781 (=NIBRBA0000114092) was isolated fermentation, arginine dihydrolase, urease and esculin from a tidal flat sample, Sinan, Korea. hydrolysis. D­glucose, L­arabinose, D­mannose, D­man­ nitol, N­acetyl­glucosamine, D­maltose, potassium glu­ Description of Isoptericola nanjingensis RS5-5_B conate, adipic acid, malic acid, trisodium citrate and phe­ Cells are Gram-staining-positive, non-flagellated and nylacetic acid are utilized. Dose not utilize capric acid. coccoid­shaped. Colonies are circular, entire, smooth and Strain RK 4Y 5­1 (=NIBRBA0000113994) was isolated light yellow­colored after 2 days on R2A at 30°C. Posi­ from a sample of ginseng field, Iljuk, Anseong, Korea. tive reactions are obtained for nitrate reduction, esculin hydrolysis and β-galactosidase in API 20NE, and nega­ Description of Arthrobacter scleromae mNW18 tive reactions are obtained for indole production, glucose Cells are Gram-staining-positive, flagellated, non-pig­ fermentation, arginine dihydrolase, urease and gelati­ mented and coccoid­shaped. Colonies are circular and nase. D­glucose, L­arabinose, D­mannose, D­mannitol, cream-colored after 3 days on minimal meduim at 25℃. N­acetyl­glucosamine, D­maltose and potassium glu­ Positive reactions are obtained for glucose fermentation, conate are utilized. Does not utilize capric acid, adipic urease, esculin hydrolysis, gelatinase and β-galactosi­ acid, malic acid, trisodium citrate and phenylacetic acid. dase in API 20NE, and negative reactions are obtained Strain RS5­5_B ( =NIBRBA0000113943) was isolat­ for nitrate reduction, indole production and arginine ed from a sample of ginseng field, Iljuk, Anseong, Ko­ dihydrolase. L­arabinose, D­mannitol, N­acetyl­glucos­ rea. amine, D­maltose, potassium gluconate, trisodium ci­ trate and phenylacetic acid are utilized. Does not utilize Description of Cellulosimicrobium funkei NS3-4_B D­glucose, D­mannose, capric acid, adipic acid and ma­ Cells are Gram-staining-positive, non-flagellated and lic acid. Strain mNW18 ( =NIBRBA0000113977) was coccoid­shaped. Colonies are punctiform, entire, smooth isolated from a forest soil sample, Gwanak Mountain, and light yellow­colored after 2 days on NA at 30°C. Seoul, Korea. Positive reactions are obtained for nitrate reduction, esculin hydrolysis and β-galactosidase in API 20NE, Description of Kocuria rosea HR39 negative reactions are obtained for indole production, Cells are Gram-staining-positive, non-flagellated and glucose fermentation, arginine dihydrolase, urease and rod­shaped. Colonies are irregular, convex, glistening gelatinase. D­glucose, L­arabinose, D­mannose, N­ace­ and pale orange/yellow­colored after 3 days on R2A at tyl­glucosamine, D­maltose and potassium gluconate are 25℃. Positive reactions are obtained for nitrate reduc­ utilized. Does not utilize D­mannitol, capric acid, adipic tion and β-galactosidase in API 20NE, and negative re­ acid, malic acid, trisodium citrate and phenylacetic acid. actions are obtained for indole production, glucose fer­ Strain NS3­4_B ( =NIBRBA0000113954) was isolated mentation, arginine dihydrolase, urease, esculin hydroly­ from a sample of ginseng field, Iljuk, Anseong, Korea. sis and gelatinase. D­glucose, L­arabinose, D­mannitol, D­maltose, potassium gluconate, adipic acid and malic Description of Microlunatus soli Gsoil 950 acid are utilized. Does not utilize D­mannose, N­ace­ Cells are Gram-staining-positive, non-flagellated and tyl­glucosamine, capric acid, trisodium citrate and phe­ rod­shaped. Colonies are circular, raised, entire and nylacetic acid. Strain HR39 ( =NIBRBA0000113993) yellow-colored after 2 days on R2A at 25℃. Positive was isolated from a tidal flat sample, Hwango-ri, Anmy­ reactions are obtained for nitrate reduction, glucose fer­ eon­eup, Taean, Korea. mentation, urease, esculin hydrolysis, gelatinase and β-galactosidase in API 20NE, and negative reactions are Description of Micrococcus flavus HME8781 obtained for indole production and arginine dihydro­ Cells are Gram-staining-positive, non-flagellated and lase. D­glucose, D­mannose, D­mannitol, N­acetyl­glu­ rod­shaped. Colonies are circular, convex, entire and cosamine, D­maltose and trisodium citrate are utilized. yellow-colored after 2 days on MA at 30℃. Negative re­ Does not utilize L­arabinose, potassium gluconate, cap­ actions are obtained for nitrate reduction, indole produc­ ric acid, adipic acid, malic acid, and phenylacetic acid. tion, glucose fermentation, arginine dihydrolase, urease, Strain Gsoil 950 (=NIBRBA0000113889) was isolated esculin hydrolysis, gelatinase and β-galactosidase in API from a sample of ginseng field, Pocheon, Korea. 20NE. D­glucose D­maltose and malic acid are utilized. February 2016 CHOI ET AL.­UNRECORDED ACTINOBACTERIA SPECIES 11

Description of Streptomyces avellaneus UKS-24 acknowledgeMents Cells are Gram-staining-positive, non-flagellated and This study was supported by the research grant “The rod­shaped. Colonies are circle, raised, entire and yel­ Survey of Korean Indigenous Species” from the Nation­ low-colored after 2 days on R2A at 25℃ Positive reac­ al Institute of Biological Resources of the Ministry of tion is obtained for β-galactosidase in API 20NE, and Environment in Korea. negative reactions are obtained for nitrate reduction, in­ dole production, glucose fermentation, arginine dihydro­ lase, urease, esculin hydrolysis and gelatinase. D­glu­ cose, L­arabinose, N­acetyl­glucosamine, potassium glu­ references conate and malic acid are utilized. Does not utilize D­mannose, D­mannitol, D­maltose, capric acid, adip­ Amann, R.I., W. Ludwig and K.H. Schleifer. 1995. Phyloge­ ic acid, trisodium citrate and phenylacetic acid. Strain netic identification and in situ detection of individual mi­ UKS­24 ( =NIBRBA0000113885) was isolated from a crobial cells without cultivation. Microbiol Rev 59:143­ wetland sample, Ungok­ri, Gochang, Korea. 169. Baldacci, E. and A. Grein. 1966. Streptomyces avellaneus Description of Streptomyces omiyaensis MS5-13 and Streptomyces libani: two new species characterized by a hazel­nut brown (avellaneus) aerial mycelium. Giorn Cells are Gram-staining-positive, non-flagellated and Microbiol 14:185­198. rod­shaped. Colonies are irregular, undulate, smooth Behrendt, U., A. Ulrich, P. Schumann, D. Naumann and K.­i. and light yellow-colored after 2 days on MA at 30℃. Suzuki. 2002. Diversity of grass­associated Microbac- Positive reactions are obtained for nitrate reduction, teriaceae isolated from the phyllosphere and litter layer esculin hydrolysis and β-galactosidase in API 20NE, after mulching the sward; polyphasic characterization of and negative reactions are obtained for indole produc­ Subtercola pratensis sp. nov., Curtobacterium herbarum tion, glucose fermentation, arginine dihydrolase, urease sp. nov. and Plantibacter flavus gen. nov., sp. nov. Int J and gelatinase. D­glucose, L­arabinose, D­mannose, D­ Syst Evol Microbiol 52:1441­1454. mannitol, N­acetyl­glucosamine, D­maltose, potassium Brown, J.M., A.G. Steigerwalt, R.E. Morey, M.I. Daneshvar, gluconate, adipic acid, malic acid and phenylacetic acid L.­J. Romero and M.M. McNeil. 2006. Characterization are utilized. Does not utilize capric acid and trisodium ci­ of clinical isolates previously identified as Oerskovia tur­ trate. Strain MS5­13 (=NIBRBA0000113953) was iso­ bata: proposal of Cellulosimicrobium funkei sp. nov. and lated from a sample of ginseng field, Anseong, Korea. emended description of the genus Cellulosimicrobium. Int J Syst Evol Microbiol 56:801­804. Description of Streptomyces atratus MA10 Buczolits, S., P. Schumann, G. Weidler, C. Radax and H.­J. Busse. 2003. Brachybacterium muris sp. nov., isolated Cells are Gram-staining-negative, non-flagellated and from the liver of a laboratory mouse strain. Int J Syst rod­shaped. Colonies are circle and yellow­colored after Evol Microbiol 53:1955­1960. 2 days on MA at 25°C. Strain MA10 ( =NIBRBA00 Chater, K.F. 2006. Streptomyces inside­out: a new perspec­ 00113970) was isolated from a forest soil sample, Gwa­ tive on the bacteria that provide us with antibiotics. Phi­ nak Mountain, Seoul, Korea. los Trans R Soc Lond B Biol Sci 361:761­768. Chater, K.F., S. Biró, K.J. Lee, T. Palmer and H. Schrempf. Description of Microbispora rosea subsp. rosea 2010. The complex extracellular biology of Streptomy­ UKS-23 ces. FEMS Microbiol Rev 34:171­198. Cells are Gram-staining-negative, non-flagellated and Conn, H.J. and I. Dimmick. 1947. Soil Bacteria Similar in rod­shaped. Colonies are circular, raised, entire and yel­ Morphology to Mycobacterium and Corynebacterium. J low-colored after 2 day on R2A at 25℃. Positive reac­ Bacteriol 54:291­303. tion is obtained for esculin hydrolysis in API 20NE, and Felsenstein, J. 1981. Evolutionary trees from DNA sequenc­ negative reactions are obtained for nitrate reduction, in­ es: a maximum likelihood approach. Journal of molecu­ dole production, glucose fermentation, arginine dihydro­ lar evolution 17:368­376. lase, urease, gelatinase and β-galactosidase. D-glucose, Fierer, N. and R.B. Jackson. 2006. The diversity and bioge­ L­arabinose, D­mannose, D­mannitol, N­acetyl­glu­ ography of soil bacterial communities. Proc Natl Acad cosamine, D­maltose, potassium gluconate, adipic acid Sci U S A 103:626­631. and malic acid are utilized. Does not utilize capric acid, Gao, B. and R.S. Gupta. 2012. Phylogenetic Framework and trisodium citrate and phenylacetic acid. Strain UKS­23 Molecular Signatures for the Main Clades of the Phylum ( =NIBRBA0000113886) was isolated from a wetland Actinobacteria. Microbiol Mol Biol Rev 76:66­112. sample, Ungok­ri, Gochang, Korea. Goodfellow, M. and G. Alderson. 1977. The Actinomy­ 12 JOURNAL OF SPECIES RESEARCH Vol. 5, No. 1

cete­genus Rhodococcus: A Home for the ‘rhodochrous’ V. Křen. 2009. Biodegradation potential of the genus Complex. J Gen Microbiol 100:99­122. Rhodococcus. Environ Int 35:162­177. Hahn, M.W. 2009. Description of seven candidate species Miyadoh, S., S. Amano, H. Tohyama and T. Shomura. 1990. affiliated with the phylum Actinobacteria, representing A taxonomic review of the genus Microbispora and a planktonic freshwater bacteria. Int J Syst Evol Microbiol proposal to transfer two species to the genus Actinomad- 59:112­117. ura and to combine ten species into Microbispora rosea. Hopwood, D.A. 2007. Streptomyces in nature and medicine. J Gen Microbiol 136:1905­1913. Oxford University Press, New York, NY. Nimaichand, S., S. Sanasam, L.­Q. Zheng, W.­Y. Zhu, L.­ Huang, Y., N. Zhao, L. He, L. Wang, Z. Liu, M. You and L. Yang, S.­K. Tang, D. S. Ningthoujam, and W.­J. Li. F. Guan. 2005. Arthrobacter scleromae sp. nov. Isolat­ 2013. Rhodococcus canchipurensis sp. nov., an actino­ ed from Human Clinical Specimens. J Clin Microbiol mycete isolated from a limestone deposit site. Int J Syst 43:1451­1455. Evol Microbiol 63:114­118. Huang, Z., X.­F. Sheng, F. Zhao, L.­Y. He, J. Huang and Q. Oren, A. and G.M. Garrity. 2014. Then and now: a systemat­ Wang. 2012. Isoptericola nanjingensis sp. nov., a min­ ic review of the systematics of prokaryotes in the last 80 eral­weathering bacterium. Int J Syst Evol Microbiol years. Antonie van Leeuwenhoek 106:43­56. 62:971­976. Parte, A.C. 2014. LPSN­list of prokaryotic names with Jukes, T.H. and C.R. Cantor. 1969. Mammalian Protein Me­ standing in nomenclature. Nucleic Acids Res 42:D613­ tabolism in Evolution of protein molecules. H. N. Munro 616. ed. Academic Press, New York. Rivas, R., M.E. Trujillo, P.F. Mateos, E. Martínez­Molina Kämpfer, P., O. Terenius, J.M. Lindh and I. Faye. 2006. Jan- and E. Velázquez. 2004. Cellulomonas xylanilytica sp. ibacter anophelis sp. nov., isolated from the midgut of nov., a cellulolytic and xylanolytic bacterium isolat­ Anopheles arabiensis. Int J Syst Evol Microbiol 56:389­ ed from a decayed elm tree. Int J Syst Evol Microbiol 392. 54:533­536. Kämpfer, P., C.­C. Young, H.­J. Busse, J.­N. Chu, P. Saitou, N. and M. Nei. 1987. The neighbor­joining method: Schumann, A.B. Arun, F.­T. Shen and P.D. Rekha. 2010. a new method for reconstructing phylogenetic trees. Mol Microlunatus soli sp. nov., isolated from soil. Int J Syst Biol Evol 4:406­425. Evol Microbiol 60:824­827. Schumann, P., F.A. Rainey, J. Burghardt, E. Stackebrandt Kageyama, A., A. Matsumoto, S. Ōmura and Y. Takahashi. and N. Weiss. 1999. Reclassification of Brevibacterium 2008. Humibacillus xanthopallidus gen. nov., sp. nov. Int oxydans (Chatelain and Second 1966) as Microbacterium J Syst Evol Microbiol 58:1547­1551. oxydans comb. nov. Int J Syst Bacteriol 49:175­177. Kageyama, A., Y. Takahashi, and S. Ōmura. 2006. Microbac- Shibata, M., E. Higashide, H. Yamamoto and K. Nakazawa. terium deminutum sp. nov., Microbacterium pumilum sp. 1962. Studies on Streptomycetes. Part I. Streptomyces nov. and Microbacterium aoyamense sp. nov. Int J Syst atratus nov. sp., producing new antituberculous antibi­ Evol Microbiol 56:2113­2117. otics rugomycin A and B. Agricultural and Biological Kim, O.­S., Y.­J. Cho, K. Lee, S.­H. Yoon, M. Kim, H. Na, Chemistry 26:228­233. S.­C. Park, Y.S. Jeon, J.­H. Lee, H. Yi, S. Won and J. Stackebrandt, E., C. Koch, O. Gvozdiak and P. Schumann. Chun. 2012. Introducing EzTaxon­e: a prokaryotic 16S 1995. Taxonomic Dissection of the Genus Micrococcus: rRNA gene sequence database with phylotypes that re­ Kocuria gen. nov., Nesterenkonia gen. nov., Kytococcus pre sent uncultured species. Int J Syst Evol Microbiol gen. nov., Dermacoccus gen. nov., and Micrococcus 62:716­721. Cohn 1872 gen. emend. Int J Syst Bacteriol 45:682­692. Liu, X.­Y., B.­J. Wang, C.­Y. Jiang and S.­J. Liu. 2007. Mi- Takeuchi, M. and K. Hatano. 1998. Union of the genera Mi- crococcus flavus sp. nov., isolated from activated sludge crobacterium Orla­Jensen and Aureobacterium Collins in a bioreactor. Int J Syst Evol Microbiol 57:66­69. et al. in a redefined genus Microbacterium. Int J Syst Ludwig, W., and H.P. Klenk. 2005. Overview: a phylogenet­ Bacteriol 48:739­747. ic backbone and taxonomic framework for procaryotic Takeuchi, M., K. Hatano, I. Sedlácek and Z. Pácová. 2002. systematics. Pages 49­65 in Bergey’s Manual of System­ Rhodococcus jostii sp. nov., isolated from a medieval atic Bacteriology. Brenner, Krieg, Staley, and Garrity grave. Int J Syst Evol Microbiol 52:409­413. eds. Springer, New York. Tamura, K., G. Stecher, D. Peterson, A. Filipski and S. Ku­ Madhaiyan, M., S. Poonguzhali, J.­S. Lee, M. Senthilkumar, mar. 2013. MEGA6: molecular evolutionary genetics K.C. Lee and S. Sundaram. 2010. Leifsonia soli sp. nov., analysis version 6.0. Mol Biol Evol 30:2725­2729. a yellow­pigmented actinobacterium isolated from teak Thompson, J.D., T.J. Gibson, F. Plewniak, F. Jeanmougin rhizosphere soil. Int J Syst Evol Microbiol 60:1322­ and D.G. Higgins. 1997. The CLUSTAL_X windows 1327. interface: flexible strategies for multiple sequence align­ Martínková, L., B. Uhnáková, M. Pátek, J. Nešvera and ment aided by quality analysis tools. Nucleic Acids Res February 2016 CHOI ET AL.­UNRECORDED ACTINOBACTERIA SPECIES 13

25:4876­4882. Yassin, A.F., H. Hupfer and K.P. Schaal. 2006. Dietzia cin- Umezawa, H., T. Tazaki, Y. Okami and S. Fukuyama. 1950. namea sp. nov., a novel species isolated from a perianal On the new source of chloromycetin, Streptomyces omi- swab of a patient with a bone marrow transplant. Int J yaensis. J Antibiotics 3:292­296. Syst Evol Microbiol 56:641­645. Ventura, M., C. Canchaya, G.F. Fitzgerald, R.S. Gupta and D. Yassin, A.F., F.A. Rainey, J. Burghardt, H. Brzezinka, S. van Sinderen. 2007. Genomics as a means to understand Schmitt, P. Seifert, O. Zimmermann, H. Mauch, D. Gi­ bacterial phylogeny and ecological adaptation: the case erth, I. Lux and K.P. Schaal. 1997. Tsukamurella tyrosi- of bifidobacteria. Antonie van Leeuwenhoek 91:351-372. nosolvens sp. nov. Int J Syst Bacteriol 47:607­614. Whitman, W.B., D.C. Coleman and W.J. Wiebe. 1998. Pro­ Zhang, J., Y. Zhang, C. Xiao, Z. Liu and M. Goodfellow. karyotes: the unseen majority. Proc Natl Acad Sci U S A 2002. Rhodococcus maanshanensis sp. nov., a novel ac­ 95:6578­6583. tinomycete from soil. Int J Syst Evol Microbiol 52:2121­ Willumsen, P., U. Karlson, E. Stackebrandt and R.M. Kro­ 2126. ppenstedt. 2001. Mycobacterium frederiksbergense sp. Zhi, X.­Y., W.­J. Li and E. Stackebrandt. 2009. An update of nov., a novel polycyclic aromatic hydrocarbon­degrad­ the structure and 16S rRNA gene sequence-based defi­ ing Mycobacterium species. Int J Syst Evol Microbiol nition of higher ranks of the class Actinobacteria, with 51:1715­1722. the proposal of two new suborders and four new families Xiao, J., Y. Luo, S. Xie and J. Xu. 2011. Serinicoccus pro- and emended descriptions of the existing higher taxa. Int fundi sp. nov., an actinomycete isolated from deep­sea J Syst Evol Microbiol 59:589­608. sediment, and emended description of the genus Serini- Zlamala, C., P. Schumann, P. Kämpfer, R. Rosselló­Mora, coccus. Int J Syst Evol Microbiol 61:16­19. W. Lubitz and H.­J. Busse. 2002. Agrococcus baldri sp. Yam, K.C., S. Okamoto, J.N. Roberts and L.D. Eltis. 2011. nov., isolated from the air in the ‘Virgilkapelle’ in Vien­ Adventures in Rhodococcus­from steroids to explosives. na. Int J Syst Evol Microbiol 52:1211­1216. Can J Microbiol 57:155­168. Yamada, K. and K. Komagata. 1972. Taxonomic studies on Submitted: May 20, 2015 coryneform bacteria. V. Classification of coryneform Revised: August 18, 2015 bacteria. J Gen Appl Microbiol 18:417­431. Accepted: February 16, 2016