Journal of Research 10(1):23-45, 2021

Description of unrecorded bacterial species belonging to the phylum in Korea

Mi-Sun Kim1, Seung-Bum Kim2, Chang-Jun Cha3, Wan-Taek Im4, Won-Yong Kim5, Myung-Kyum Kim6, Che-Ok Jeon7, Hana Yi8, Jung-Hoon Yoon9, Hyung-Rak Kim10 and Chi-Nam Seong1,*

1Department of Biology, Sunchon National University, Suncheon 57922, Republic of Korea 2Department of Microbiology, Chungnam National University, Daejeon 34134, Republic of Korea 3Department of Biotechnology, Chung-Ang University, Anseong 17546, Republic of Korea 4Department of Biotechnology, Hankyong National University, Anseong 17579, Republic of Korea 5Department of Microbiology, College of Medicine, Chung-Ang University, Seoul 06974, Republic of Korea 6Department of Bio & Environmental Technology, Division of Environmental & Life Science, College of Natural Science, Seoul Women’s University, Seoul 01797, Republic of Korea 7Department of Life Science, Chung-Ang University, Seoul 06974, Republic of Korea 8School of Biosystem and Biomedical Science, Korea University, Seoul 02841, Republic of Korea 9Department of Food Science and Biotechnology, Sungkyunkwan University, Suwon 16419, Republic of Korea 10Department of Laboratory Medicine, Saint Garlo Medical Center, Suncheon 57931, Republic of Korea

*Correspondent: [email protected]

For the collection of indigenous prokaryotic species in Korea, 77 strains within the phylum Actinobacteria were isolated from various environmental samples, fermented foods, animals and clinical specimens in 2019. Each strain showed high 16S rRNA gene sequence similarity (>98.8%) and formed a robust phylogenetic clade with actinobacterial species that were already defined and validated with nomenclature. There is no official description of these 77 bacterial species in Korea. The isolates were assigned to 77 species, 31 genera, 18 families, 14 orders and 2 classes of the phylum Actinobacteria. All the strains except one Coriobacteriia strain were affiliated within the class . Among them, the orders Streptomycetales and Microbacteriales were predominant. A number of strains were isolated from forest soils, riverside soils, and ginseng cultivated soils. Twenty-nine strains were isolated from ‘Protected Ecosystem and Scenery Areas’. Morphological properties, basic biochemical characteristics, isolation source and strain IDs are described in the species descriptions. Keywords: ‌16S rRNA gene sequence, Actinobacteria, unrecorded species

Ⓒ 2021 National Institute of Biological Resources DOI:10.12651/JSR.2021.10.1.023

Introduction species are relatively abundant in terrestrial and aquatic environments where they are involved in the decomposi­ The phylum Actinobacteria is one of the largest groups tion and recycling of organic matter (Servin et al., 2008). in the (Goodfellow, 2012). In recent In addition to their saprophytic property, several genera, years, a proper hierarchical classification system for this such as , (Dangle et al., group was established based on genome analysis. Conse­ 2019) and Clavibacter (Hwang et al., 2019), are patho­ quently, on August 2020, the phylum Actinobacteria con­ genic to animals (including humans) and plants (Qin et al., sisted of 6 classes, 46 orders, 78 families and more than 2011). Also, endophytic actinobacteria have been isolated 400 genera [LPSN (https://www.bacterio.net/); Salam et from a variety of healthy plants (Qin et al., 2011). al., 2020]. In Korea, 329 species with valid names were isolated Members of the phylum Actinobacteria showed great from various natural environments, fermented foods, diversity in terms of their habitat, morphology and physi­ waste­water, compost and clinical specimens [LPSN (https:// ology­ (Goodfellow and Williams, 1983). Actinobacterial www.bacterio.net/); Bae et al., 2016]. Moreover, 249 un­ 24 JOURNAL OF SPECIES RESEARCH Vol. 10, No. 1 recorded species were also discovered up to 2018 (Choi et senstein, 1981) and the maximum parsimony (Fitch, 1971) al., 2016; Kim et al., 2016; 2017; 2019; Ko et al., 2017; methods with the MEGA 6.0 (Tamura et al., 2013) with Lee et al., 2018). bootstrap values based on 1,000 replicates (Felsenstein, In 2019, the authors isolated a great number of unrec­ 1985). orded prokaryotic species from diverse environmental samples, artificial sources and clinical specimens in Korea. In particular, included in the isolation sources were nine Results and Discussion ‘Protected Ecosystem and Scenery Areas’ designated by the Ministry of Environment (http://www.cbd-chm.go.kr/). All the 77 strains belonged to the phylum Actinobacte- The present report focuses on the description of unrecorded ria and were affiliated with 2 classes, 14 orders, 18 fam­ species belonging to the phylum Actinobacteria which ilies and 31 genera (Table 1). All the strains except one previously had not been isolated in Korea. Here we report were affiliated with the class Actinomycetia. Seventy-six 77 unrecorded actinobacterial strains in Korea. strains were affiliated with 13 orders:Streptomycetales (29 strains), Microbacteriales (14 strains), (8 strains), (6 strains), Micromonosporales (4

Materials and Methods strains), (3 strains), Brevibacteriales (3 strains), Cellulomonadales (2 strains), Pseudonocardi- A total of 77 bacterial strains assigned to the phylum ales (2 strains), Bifidobacteriales, Bogoriellales, Derma­ Actinobacteria were isolated from various environmental bacterales and Dermatophilales (each 1 strain). All the samples including forest soils, tidal sediments, seashore Streptomycetales, Microbacteriales, Micrococcales, Micro- sands, cave soils, ginseng cultivated soil, fermented foods monosporales, Propionibacteriales, Brevibacteriales and clinical specimens in 2019 (Table 1). Each sample was strains belonged to the single families , processed separately, spread onto diverse culture agar , , ­ media (Becton Dickinson) such as anaerobe basal (AB), ceae, , , Pseudonocar- blood (BA), international project medium 7 diaceae, , , Dermabac- (ISP7), Luria broth (LB), Mueller-Hinton (MH), marine teraceae and , respectively. The strains (MA), Reasoner’s 2A (R2A) and tryptic soy (TSA), and belonging to the order Mycobacteriales were affiliated incubated at 20-37℃ for 1-12 days. All strains were within four families: (5 strains), Dietziaceae, puri­fied as single colonies and stored as 10-20% glycerol and (each 1 strain). Two suspension at -80℃ as well as lyophilized ampoules. families Actinotaleaceae and Oerskoviaceae were found Colony morphology of the strains was observed on in the order Cellulomonadales. agar plates with a magnifying lens after cells grew up to Only one isolate belonged to the family Coriobacteria- stationary phase. Cellular morphology and cell size were ceae within the class Coriobacteriia. examined by either transmission electron microscopy or The strains were isolated from diverse sources: 64 scanning electron microscopy (Figs. 1 & 2). Biochemical strains from soil including forest soils, riverside soils, characteristics were tested using API 20NE (for aerobic ginseng-cultivated soils, meadow soils, and cave soils; 7 isolates) or API 20A (for anaerobic strains LPB0331 and strains from tidal flat sediments or seashore sands; 2 strains LPB0332) galleries (bioMérieux) according to the manu­ each from animal intestines and fermented foods and one facturer’s instructions. strain each from clinical specimen and seawater. Geo­ DNA extraction, PCR amplification and 16S rRNA gene graphic regions of the strains were as follows: 28 strains sequencing were carried out as described previously (Chun from Gangwon Province; 16 strains from Gyeongsangbuk and Goodfellow, 1995). The 16S rRNA gene sequences­ Province; 12 strains from Jeollanam Province; 4 strains of the strains assigned to the phylum Actinobacteria each from Daejeon, Jeju and Chungcheongnam Province; were compared with the sequences held in GenBank by 3 strains from Gyeonggi Province; 2 strains each from BLASTN and also analyzed using the EzTaxon-e server Seoul and Incheon and one strain each from Sejong and (http://www.ezbiocloud.net/; Yoon et al., 2017). For Chungcheongbuk Province. In particular, 29 strains were phylogenetic analyses, multiple alignments were per­ isolated from ‘Protected Ecosystem and Scenery Areas’. formed using the ClustalW program (Thompson et al., All isolates were Gram-stain-positive and chemohetero­ 1994) and gaps were edited in the BioEdit program (Hall, trophic. Figure 3 shows the phylogenetic assignment of 1999). Evolutionary distances were calculated using the the strains based on 16S rRNA gene sequences. Jukes-Cantor model (Jukes and Cantor, 1969). The phylo­ Here we report the 77 unrecorded bacterial species in genetic trees were constructed by using the neighbor-join­ Korea belonging to the phylum Actinobacteria. ing (Saitou and Nei, 1987), the maximum likelihood (Fel­ February 2021 Kim et al. Unrecorded Actinobacteria species of Korea 25

25°C, 3d 25°C, 3d 25°C, 3d 25°C, 3d 30°C, 3d 25°C, 3d 30°C, 4d 30°C, 4d 30°C, 3d 30°C, 4d 30°C, 4d 25°C, 3d 30°C, 3d 25°C, 3d 25°C, 3d 30°C, 4d 30°C, 4d 25°C, 3d 25°C, 3d 25°C, 3d 25°C, 3d 25°C, 3d 30°C, 4d 30°C, 4d 30°C, 3d condition Incubation R2A R2A R2A R2A AB R2A MH MH R2A MH MH R2A MA R2A R2A MH MH R2A R2A TSA R2A R2A MH MH R2A Medium Isolation † † † † † † † † † † † R.S./Gw G.S./Gb G.S./Gb G.S./Gb G.S./Gb M.S./Gw A.I./CDS G.S./Gb R.S./Gw R.S./Gw R.S./Gw R.S./Gw G.S./Gb T.F./GSI G.S./Gb F.F./GSI R.S./Gw R.S./Gw F.S./Gb S.S./CDS F.S./Gb G.S./Gb T.F./CDS R.S./Gw M.S./Gw Source/Region

99.0 98.9 98.8 98.9 99.7 99.5 99.9 99.6 99.8 99.6 99.4 99.9 99.2 98.8 99.6 99.8 99.1 99.0 99.2 99.1 99.6 (%) 100.0 100.0 100.0 100.0 Similarity . longum subsp Most closely related species A. ferrariae B. avium B. sandarakinum B. siliguriense G. satyanarayanai P. soli P. B. longum B. alimentarium A. citreus A. tumbae O. jenensis A. versicolor A. fucosus A. mangrove L. massiliensis M. flavum M. invictum M. lemovicicum M. murale M. nanhaiense M. shaanxiense M. thalassium M. trichothecenolyticum M. manganoxydans ilicis P. NIBR ID . NIBRBAC000503262 NIBRBAC000503403 NIBRBAC000503413 NIBRBAC000503412 NIBRBAC000503416 NIBRBAC000503224 NIBRBAC000503355 NIBRBAC000503405 NIBRBAC000503254 NIBRBAC000503267 NIBRBAC000503256 NIBRBAC000503404 NIBRBAC000503252 NIBRBAC000503414 NIBRBAC000503401 NIBRBAC000503259 NIBRBAC000503253 NIBRBAC000503304 NIBRBAC000503354 NIBRBAC000503305 NIBRBAC000503417 NIBRBAC000503329 NIBRBAC000503269 NIBRBAC000503223 NIBRBAC000503263 Actinobacteria Strain ID 19D1G4 G9 G37 G24 N20 13H - 3 LPB0331 G56 19D1A19 19D2A1 19D1A72 G36 CAU 1605 N14 KR3 19D1C16 19D1A9 SO98 LPB0322 SO111 N40 BSSP - R25 19D2C13 13H - 2 19D1L6 Genus Georgenia Pedococcus Bifidobacterium Brachybacterium Oerskovia Family Actinotaleaceae Brevibacteriaceae Bogoriellaceae Intrasporangiaceae Bifidobacteriaceae Microbacteriaceae Micrococcaceae Oerskoviaceae The taxonomic affiliations of isolates belonging to the phylum Order* Cellulomonadales Brevibacteriales Bogoriellales Dermatophilales Bifidobacteriales Table 1. Table Dermabacterales Microbacteriales Micrococcales 26 JOURNAL OF SPECIES RESEARCH Vol. 10, No. 1

30°C, 3d 25°C, 3d 25°C, 3d 25°C, 3d 25°C, 3d 25°C, 3d 25°C, 5d 30°C, 3d 25°C, 3d 30°C, 4d 30°C, 2d 37°C, 3d 25°C, 2d 30°C, 3d 30°C, 3d 25°C, 3d 25°C, 3d 30°C, 4d 30°C, 4d 30°C, 4d 25°C, 4d 25°C, 3d 30°C, 4d 25°C, 3d condition Incubation R2A TSA R2A R2A R2A R2A MA MA R2A MH MA BA MA R2A ISP7 R2A R2A MH MH MH TSA R2A MH R2A Medium Isolation † † † † † † † † F.S./Jn F.S./GSI F.S./Gb R.S./Gw F.S./Gw F.F./GSI F.S./Jn F.S./Gb R.S./Gw T.F./CDS R.S./Gw T.F./CDS F.S./Gw T.F./GSI H.S./GSI S.W./Jn F.S./CDS G.S./Gb F.S./Jj R.S./Gw R.S./Gw F.S./Jn G.S./Gb F.S./Jj Source/Region

99.9 98.9 99.5 99.1 99.2 99.0 99.5 98.9 99.6 99.7 99.1 99.3 99.9 99.0 99.6 99.4 99.8 99.7 99.4 99.1 (%) 100.0 100.0 100.0 100.0 Similarity . subsp Most closely related species K. purpeofusca K. xanthocidica S. carbonis P. phenanthrenivorans P. L. urticae D. lutea limnetica W. M. wolinskyi N. alba A. marinum M. aquaticus L. albidocapillata albidocapillata L. guizhouensis P. polychromogenes P. R. aeria R. amarae M. avicenniae M. oryzae M. schwarzwaldensis N. grenadensis N. nova N. tengchongensis R. artemisiae N. phosphati NIBR ID . NIBRBAC000503341 NIBRBAC000503409 NIBRBAC000503382 NIBRBAC000503260 NIBRBAC000503258 NIBRBAC000502986 NIBRBAC000503218 NIBRBAC000503402 NIBRBAC000503294 NIBRBAC000503408 NIBRBAC000503270 NIBRBAC000503336 NIBRBAC000503328 NIBRBAC000503335 NIBRBAC000503349 NIBRBAC000503307 NIBRBAC000503392 NIBRBAC000503406 NIBRBAC000503002 NIBRBAC000503264 NIBRBAC000503272 NIBRBAC000503295 NIBRBAC000503418 NIBRBAC000502998 Actinobacteria Strain ID LPB0280 S36 MMS19 - T35 19D1F19 19D1C14 BT46 R21 SR3 FS100 S5 19D2V10 BSSP - M28 BSSP - M29 JBTF - M16 LPB0310 KYW1971 R_77 G92 BT360 19D1S1 19D1V24 JDB110 R12 BT343 Genus Streptacidiphilus Williamsia Micromonospora Family Streptomycetaceae Micrococcaceae Dietziaceae Gordoniaceae Mycobacteriaceae Nocardiaceae Nocardioidaceae Continued. The taxonomic affiliations of isolates belonging to the phylum Order* Streptomycetales Micrococcales Table 1. Table Table 1. Table Micromonosporales Mycobacteriales Propionibacteriales Pseudonocardiales February 2021 Kim et al. Unrecorded Actinobacteria species of Korea 27

30°C, 3d 30°C, 3d 25°C, 3d 37°C, 3d 30°C, 4d 25°C, 3d 30°C, 3d 25°C, 4d 30°C, 4d 30°C, 3d 30°C, 3d 30°C, 3d 30°C, 3d 30°C, 3d 30°C, 4d 37°C, 3d 25°C, 7d 25°C, 3d 30°C, 4d 25°C, 5d 37°C, 3d 30°C, 3d 30°C, 4d 30°C, 3d 30°C, 3d 25°C, 3d 30°C, 3d 30°C, 4d condition Incubation AB TSA R2A R2A MH 1/10LB R2A R2A MH R2A R2A R2A R2A R2A MH NA TSA R2A MH R2A R2A R2A MH R2A R2A R2A R2A MH Medium Isolation † † † † † † † † † † † † † R.S./Jn A.I./CDS F.S./Jn F.S./Jj R.S./Gw R.S./Gw F.S./Gw F.S./Gw F.S./Gb R.S./Jn F.S./Jn C.S./Gw C.S./Gw M.S./Gw R.S./Gw S.S./GSI F.S./Jj F.S./Gb R.S./Gw F.S./Jn R.S./Jn F.S./CDS R.S./Gw F.S./CDS F.S./Jn F.S./Cb F.S./Jn R.S/Gw Source/Region

Denotes the ‘Protected Ecosystem and Scenery Areas.’ Denotes the ‘Protected Ecosystem and Scenery † 99.9 99.7 99.9 99.6 99.8 99.0 99.2 99.9 99.2 99.9 99.9 99.9 99.5 99.2 99.8 99.9 99.9 99.9 (%) 100.0 100.0 100.0 100.0 100.0 100.0 100.0 100.0 100.0 100.0 Similarity . asoensis subsp Most closely related species S. albogriseolus C. aerofaciens S. cavourensis S. coelescens S. amakusaensis S. aureus S. badius S. brevispora S. cacaoi S. corchorusii S. europaeiscabiei S. finlayi S. formicae S. fragilis S. fulvissimus S. globosus S. griseorubiginosus S. hydrogenans S. netropsis S. nigrescens S. phaeoluteichro- matogenes S. populi S. pratensis S. pseudovenezuelae S. recifensis S. tanashiensis S. virginiae S. zaomyceticus NIBR ID . . NIBRBAC000503388 NIBRBAC000503339 NIBRBAC000503282 NIBRBAC000503274 NIBRBAC000503276 NIBRBAC000502992 NIBRBAC000503221 NIBRBAC000503306 NIBRBAC000503380 NIBRBAC000503387 NIBRBAC000503371 NIBRBAC000503220 NIBRBAC000503219 NIBRBAC000503222 NIBRBAC000503277 NIBRBAC000503235 NIBRBAC000503283 NIBRBAC000503297 NIBRBAC000503278 NIBRBAC000503303 NIBRBAC000503389 NIBRBAC000503377 NIBRBAC000503275 NIBRBAC000503378 NIBRBAC000503372 NIBRBAC000503284 NIBRBAC000503384 NIBRBAC000503273 Actinomycetia Actinobacteria 111 Strain ID - R - 5 LPB0332 EAC34 19D1L39 19D2C16 BT63 9C - 1 SO100 BG138 R - 9 MMS19 - T27 5C - 2 5C - 1 13H - 1 19D2F17 CAU 1564 EAC30 SO94 19D2S3 JDB244 R - 21 DS - 12 19D1T8 F MMS19 - T31 EAC17 MMS19 - T12 19D1A31 ; the rests to class Genus Streptomyces Collinsella Coriobacteriia Family belongs to the class Streptomycetaceae Coriobacteriaceae Coriobacteriales Continued. The taxonomic affiliations of isolates belonging to the phylum Order* Streptomycetales Table 1. Table Table 1. Table *The order H.S., food; fermented F.F., intestine; animal A.I., sand; S.S., seashore sediment; flat tidal T.F., soils; C.S., cave soil; M.S., meadow soil; R.S, riverside soil; forest F.S., soil; G.S., Ginseng-cultivated source: of the Abbreviations seawater. clinical specimen; S.W., Gangwon; Gb, Gyeongbuk; Jn, Jeonnam; Jj, Jeju; CDS, Chungnam/ Daejeon/Sejong; GSI, Gyeonggi/Seoul/Incheon; Cb, Chungbuk. Abbreviations of the region: Gw, TSA, tryptic soy. 2A; Anaerobe basal; BA, Blood;.ISP7, international streptomyces project medium 7; LB, Luria broth; MH, Mueller Hinton; MA, marine; R2A, Reasoner’s AB, Abbreviations of the agar medium: Coriobacteriales 28 JOURNAL OF SPECIES RESEARCH Vol. 10, No. 1

Fig. 1. Transmission electron micrographs of cells of the isolates. Bar, 1 μm. Strains: 1, LPB0331; 2, N20; 3, G24; 4, G9; 5, G37; 6, 19D1G4; 7, 19D1L6; 8, G56; 9, 13H-3; 10, 19D1A19; 11, 19D1A72; 12, G36; 13, CAU 1605; 14, N14; 15, KR3; 16, 19D1C16; 17, 19D1A9; 18, SO98; 19, LPB0322; 20, SO111; 21, N40; 22, BSSP-R25; 23, 19D2C13; 24, 19D2A1; 25, 13H-2; 26, 19D1F19; 27, JBTF-M16; 28, LPB0310; 29, KYW1971; 30, R21; 31, R_77; 32, BT360; 33, SR3; 34, FS100; 35, S5; 36, 19D2V10; 37, 19D1S1; 38, 19D1V24; 39, JDB110; 40, R12; 41, BSSP-M28; 42, 19D1C14; 43, BT343; 44, BSSP-M29; 45, BT46; 46, MMS19-T35; 47, LPB0280; 48, R-5; 49, 19D2C16; 50, BT63; 51, 9C-1; 52, BG138; 53, EAC34; 54, 19D1L39; 55, R-9; 56, MMS19-T27; 57, 5C-2; 58, 5C-1; 59, 13H-1; 60, 19D2F17; 61, CAU 1564; 62, 19D2S3; 63, R-21; 64, DS-12; 65, 19D1T8; 66, F-111; 67, MMS19-T31; 68, EAC17; 69, MMS19-T12; 70, 19D1A31; 71, LPB0332. February 2021 Kim et al. Unrecorded Actinobacteria species of Korea 29

Fig. 1. Continued.

Fig. 2. Scanning electron micrographs of cells of the isolates. Bar, 1 μm. Strains: 1, G92; 2, S36; 3, SO100; 4, EAC30; 5, SO94; 6, JDB244.

Description of Bifidobacterium longum subsp. longum reaction for esculin hydrolysis, acid production from LPB0331 salicin (weak), glycerol, D-cellobiose, D-rhamnose and D-trehalose. In the API 20A system, negative reaction for Cells are anaerobic, Gram-staining-positive, non-flagel­ oxidase activity, indole formation, urease activity, gelatin lated and rod shaped. Colonies are circular, convex, entire hydrolysis and acid production from D-glucose, D-manni­ and cream colored after incubation for 3 days on anaerobe tol, D-lactose, sucrose, D-maltose, D-xylose, L-arabinose, basal medium at 30℃. In the API 20A system, positive D-mannose, D-melezitose, D-raffinose and D-sorbitol. 30 JOURNAL OF SPECIES RESEARCH Vol. 10, No. 1

Fig. 3. Neighbor-joining phylogenetic tree, based on 16S rRNA gene sequences, showing the relationship between the isolates and their rela­ tives of the phylum Actinobacteria. Evolutionary distances, generated using the model of Jukes & Cantor (1969), are based on 1155 unambi­ guously aligned nucleotides. Bootstrap values (>70%) are shown above nodes. Filled circles indicate the nodes recovered by three other treeing methods including maximum likelihood, maximum parsimony and neighbor-joining. Bar, 0.02 substitutions per nucleotide position. February 2021 Kim et al. Unrecorded Actinobacteria species of Korea 31

Fig. 3. Continued. 32 JOURNAL OF SPECIES RESEARCH Vol. 10, No. 1

Strain LPB0331 (=NIBRBAC000503355) was isolated Yeongju, Gyeongsangbuk Province, Korea (36°53′51.5″N from the intestine of a laboratory mouse in Daejeon, Korea 128°28′02.3″E). (36°23′56.11″N 127°23′41.76″E). Description of Brevibacterium siliguriense G37 Description of Georgenia satyanarayanai N20 Cells are aerobic, Gram-staining-positive, non-flagel­ Cells are aerobic, Gram-staining-positive, non-flagel­ lated and rod shaped. Colonies are circular, smooth and lated and rod shaped. Colonies are circular, convex, entire light orange colored after incubation for 3 days on R2A and white colored after incubation for 3 days on R2A agar agar at 25℃. In the API 20NE system, positive reaction at 25℃. In the API 20NE system, positive reaction for nit­ for nitrate reduction and utilization of D-glucose, D-man­ rate reduction, hydrolysis of esculin, oxidase activity and nose, D-mannitol, potassium gluconate, capric acid, malic utilization of D-glucose, D-mannose, N-acetyl-glucosa­ acid, trisodium citrate and phenylacetic acid. In the API mine, D-maltose and potassium gluconate. In the API 20NE system, negative reaction for indole production, 20NE system, negative reaction for indole production, glucose fermentation, arginine dihydrolase, urease activity, glucose fermentation, arginine dihydrolase, urease acti­ hydrolysis of esculin and gelatin, β-galactosidase activity, vity, hydrolysis of gelatin, β-galactosidase activity and oxidase activity and utilization of L-arabinose, N-acetyl- utilization of L-arabinose, D-mannitol, capric acid, adipic glucosamine, D-maltose and adipic acid. Strain G37 acid, malic acid, trisodium citrate and phenylacetic acid. (=NIBRBAC000503413) was isolated from ginseng-cul­ Strain N20 (=NIBRBAC000503416) was isolated from tivated soil in Yeongju, Gyeongsangbuk Province, Korea ginseng-cultivated soil in Yeongju, Gyeongsangbuk Prov­­ (36°53′51.5″N 128°28′02.3″E). ince, Korea (36°53′51.5″N 128°28′02.3″E).

Description of Brevibacterium avium G24 Description of Actinotalea ferrariae 19D1G4 Cells are aerobic, Gram-staining-positive, non-flagellat­ Cells are aerobic, Gram-staining-positive, non-flagel­ ed and rod shaped. Colonies are circular, smooth and cream lated and short rod shaped. Colonies are circular, convex, colored after incubation for 3 days on R2A agar at 25℃. glistening and yellow colored after incubation for 4 days In the API 20NE system, positive reaction for nitrate­ re­ on MH agar at 30℃. In the API 20NE system, positive re­ duction and utilization of D-glucose, L-arabinose, D-man­ action for nitrate reduction, glucose fermentation, hydro- nitol, potassium gluconate, capric acid, malic acid, triso­ lysis of esculin and gelatin, β-galactosidase activity and dium citrate and phenylacetic acid. In the API 20NE sys­ utilization of D-glucose, D-mannose, N-acetyl-glucosa­ tem, negative reaction for indole production, glucose fer­ mine (weak) and D-maltose. In the API 20NE system, mentation, arginine dihydrolase, urease activity, hydrolysis negative reaction for indole production, arginine dihydro­ of esculin and gelatin, β-galactosidase activity, oxi­dase lase, urease activity, oxidase activity and utilization of L- activity­ and utilization of D-mannose, N-acetyl-glucosa­ arabinose, D-mannitol, potassium gluconate, capric acid, mine, D-maltose and adipic acid. Strain G24 ( =NIBR adipic acid, malic acid, trisodium citrate and phenylacetic BAC000503412) was isolated from ginseng-cultivated soil acid. Strain 19D1G4 (=NIBRBAC000503262) was iso­ in Yeongju, Gyeongsangbuk Province, Korea (36°53′ lated from soil around the Dong River, Gangwon Prov­ 51.5″N 128°28′02.3″E). ince, Korea (37°22′56.8″N 128°40′01.2″E).

Description of Brevibacterium sandarakinum G9 Description of Oerskovia jenensis 19D1L6 Cells are aerobic, Gram-staining-positive, non-flagel­ Cells are facultatively aerobic, Gram-staining-positive, lated and rod shaped. Colonies are circular, smooth and flagellated and rod shaped. Colonies are circular, convex, orange colored after incubation for 3 days on R2A agar at glistening and light yellow colored after incubation for 4 25℃. In the API 20NE system, positive reaction for nit­ days on MH agar at 30℃. In the API 20NE system, posi­ rate reduction and utilization of D-glucose, D-mannitol, tive reaction for nitrate reduction, glucose fermentation, potassium gluconate, malic acid and trisodium citrate. In urease activity, esculin hydrolysis, β-galactosidase activity­ the API 20NE system, negative reaction for indole pro­ and utilization of D-glucose, L-arabinose, D-mannose, duction, glucose fermentation, arginine dihydrolase, urease N-acetyl-glucosamine, D-maltose and potassium gluco­ activity, hydrolysis of esculin and gelatin, β-galactosidase nate. In the API 20NE system, negative reaction for indole activity, oxidase activity and utilization of L-arabinose, production, arginine dihydrolase, gelatin hydrolysis, oxi­ D-mannose, N-acetyl-glucosamine, D-maltose, capric acid, dase activity and utilization of D-mannitol, capric acid, adipic acid and phenylacetic acid. Strain G9 (=NIBRBAC adipic acid, malic acid, trisodium citrate and phenylacetic 000503403) was isolated from ginseng-cultivated soil in acid. Strain 19D1L6 (=NIBRBAC000503263) was iso­ February 2021 Kim et al. Unrecorded Actinobacteria species of Korea 33 lated from soil around the Dong River, Gangwon Prov­ Description of Agrococcus versicolor 19D1A72 ′ ″ ′ ″ ince, Korea (37°22 56.8 N 128°40 01.2 E). Cells are aerobic, Gram-staining-positive, non-flagel­ lated and ovoid or short rod shaped. Colonies are circular, Description of Brachybacterium alimentarium G56 convex, glistening and orange colored after incubation Cells are aerobic, Gram-staining-positive, non-flagel­ for 4 days on MH agar at 30℃. In the API 20NE system, lated and coccoid or ovoid. Colonies are circular, smooth, positive reaction for hydrolysis of esculin and gelatin and glistening and yellow colored after incubation for 3 days β-galactosidase activity. In the API 20NE system, nega­tive on R2A agar at 25℃. In the API 20NE system, positive reaction for nitrate reduction, indole production, glucose reaction for urease activity, hydrolysis of esculin, β-galac­ fermentation, arginine dihydrolase, urease activity, oxidase tosidase activity and utilization of D-glucose, L-arabinose, activity and utilization of D-glucose, L-arabinose, D-man­ D-mannose, D-mannitol, N-acetyl-glucosamine, D-mal­ nose, D-mannitol, N-acetyl-glucosamine, D-maltose, tose and potassium gluconate. In the API 20NE system, pota­ssium gluconate, capric acid, adipic acid, malic acid, negative reaction for nitrate reduction, indole production, trisodium­ citrate and phenylacetic acid. Strain 19D1A72 glucose fermentation, arginine dihydrolase, hydrolysis (=NIBRBAC000503256) was isolated from soil around of gelatin, oxidase activity and utilization of capric acid, the Dong River, Gangwon Province, Korea (37°22′56.8″N adipic acid, malic acid, trisodium citrate and phenylacetic 128°40′01.2″E). acid. Strain G56 (=NIBRBAC000503405) was isolated Description of Agromyces fucosus G36 from ginseng-cultivated soil in Yeongju, Gyeongsangbuk Province, Korea (36°53′51.5″N 128°28′02.3″E). Cells are aerobic, Gram-staining-positive, non-flagella­ ted and rod shaped. Colonies are entire, convex and yellow Description of Pedococcus soli 13H-3 colored after incubation for 3 days on R2A agar at 25℃. In the API 20NE system, positive reaction for hydrolysis Cells are aerobic, Gram-staining-positive, non-flagel­ of esculin and utilization of L-arabinose, D-mannitol, lated and coccoid. Colonies are circular and white colored N-acetyl-glucosamine and phenylacetic acid. In the API - after incubation for 3 days on R2A agar at 20 40℃. In 20NE system, negative reaction for nitrate reduction, indole the API 20NE system, positive reaction for hydrolysis of production, glucose fermentation, arginine dihydrolase, esculin and gelatin, β-galactosidase activity and utiliza­ urease activity, hydrolysis of gelatin, β-galactosidase acti­ tion of D-glucose, L-arabinose, D-mannose, D-mannitol, vity, oxidase activity and utilization of D-glucose, D-man- N-acetyl-glucosamine, potassium gluconate, malic acid nose, D-maltose, potassium gluconate, capric acid, adipic and trisodium citrate. In the API 20NE system, negative acid, malic acid and trisodium citrate. Strain G36 (=NIBR reaction for nitrate reduction, indole production, glucose BAC000503404) was isolated from ginseng-cultivated soil fermentation, arginine dihydrolase, urease activity, oxidase­ in Yeongju, Gyeongsangbuk Province, Korea (36°53′51.5″ activity and utilization of D-maltose, capric acid, adipic N 128°28′02.3″E). acid and phenylacetic acid. Strain 13H-3 (=NIBRBAC 000503224) was isolated from meadow soil in Yeongwol, Description of Agromyces mangrovi CAU 1605 Gangwon Province, Korea (37°23′04.0″N 128°41′01.3″E). Cells are aerobic, Gram-staining-positive, non-flagel­ lated and short rod shaped. Colonies are circular, convex, Description of Agrococcus citreus 19D1A19 smooth, shiny, opaque and yellow colored after incubation Cells are aerobic, Gram-staining-positive, non-flagel­ for 2-3 days on marine agar at 30℃. In the API 20NE lated and ovoid or short rod shaped. Colonies are circular, system, positive reaction for esculin hydrolysis and β-gal­ convex, glistening and yellow colored after incubation for actosidase activity. In the API 20NE system, negative 4 days on MH agar at 30℃. In the API 20NE system, posi­ reaction for nitrate reduction, indole production, glucose tive reaction for esculin hydrolysis and β-galactosidase fermentation, arginine dihydrolase, urease activity, gela­ activity. In the API 20NE system, negative reaction for tin hydrolysis and utilization of D-glucose, L-arabi­nose, nitrate reduction, indole production, glucose fermentation, D-mannose, D-mannitol, N-acetyl-glucosamine, D-mal- arginine dihydrolase, urease activity, gelatin hydrolysis, tose, potassium gluconate, capric acid, adipic acid, malic oxidase activity and utilization of D-glucose, L-arabi­ acid, trisodium citrate and phenylacetic acid. Strain CAU nose, D-mannose, D-mannitol, N-acetyl-glucosamine, 1605 (=NIBRBAC000503252) was isolated from tidal

D-maltose, potassium gluconate, capric acid, adipic acid, flat sediment in Incheon, Korea (37°32′41.0″N 126°25′ malic acid, trisodium citrate and phenylacetic acid. Strain 53.9″E). 19D1A19 ( =NIBRBAC000503254) was isolated from soil around the Dong River, Gangwon Province, Korea Description of Leucobacter massiliensis N14 (37°22′56.8″N 128°40′01.2″E). Cells are aerobic, Gram-staining-positive, non-flagella­ 34 JOURNAL OF SPECIES RESEARCH Vol. 10, No. 1 ted and rod shaped. Colonies are circular, convex, glisten­ glistening and yellow colored after incubation for 4 days ing and yellow colored after incubation for 3 days on R2A on MH agar at 30℃. In the API 20NE system, positive agar at 25℃. In the API 20NE system, positive reaction reaction for hydrolysis of esculin and gelatin, β-galacto­ for hydrolysis of esculin, oxidase activity and utilization of sidase activity and oxidase activity. In the API 20NE sys­ D-mannitol. In the API 20NE system, negative reaction for tem, negative reaction for nitrate reduction, indole produc­ nitrate reduction, indole production, glucose fermentation, tion, glucose fermentation, arginine dihydrolase, urease arginine dihydrolase, urease activity, hydrolysis of gelatin, activity and utilization of D-glucose, L-arabinose, D-man­ β-galactosidase activity and utilization of D-glucose, nose, D-mannitol, N-acetyl-glucosamine, D-maltose, L-arabinose, D-mannose, N-acetyl-glucosamine, D-mal­ potassium gluconate, capric acid, adipic acid, malic acid, tose, potassium gluconate, capric acid, adipic acid, malic trisodium citrate and phenylacetic acid. Strain 19D1A9 acid, trisodium citrate and phenylacetic acid. Strain N14 (=NIBRBAC000503253) was isolated from soil around (=NIBRBAC000503414) was isolated from ginseng-cul­ the Dong River, Gangwon Province, Korea (37°22′56.8″N tivated soil in Yeongju, Gyeongsangbuk Province, Korea 128°40′01.2″E). (36°53′51.5″N 128°28′02.3″E). Description of Microbacterium murale SO98 Description of KR3 Cells are aerobic, Gram-staining-positive, non-flagel­ Cells are aerobic, Gram-staining-positive, non-flagel­ lated and short rod shaped. Colonies are circular, smooth, lated and ovoid or rod shaped. Colonies are circular, con­ opaque and yellow colored after incubation for 3 days on vex and yellow colored after incubation for 3 days on R2A R2A agar at 25℃. In the API 20NE system, positive reac­ agar at 25℃. In the API 20NE system, positive reaction tion for nitrate reduction, esculin hydrolysis and utilization for hydrolysis of esculin, β-galactosidase activity and uti­ of D-glucose, L-arabinose, D-mannose and D-mannitol. lization of D-glucose, D-mannose, D-mannitol, N-acetyl- In the API 20NE system, negative reaction for indole pro­ glucosamine, D-maltose, potassium gluconate, malic acid duction, glucose fermentation, arginine dihydrolase, ure­ and trisodium citrate. In the API 20NE system, negative ase activity, gelatin hydrolysis, β-galactosidase activity, reaction for nitrate reduction, indole production, glucose oxidase activity and utilization of N-acetyl-glucosamine, fermentation, arginine dihydrolase, urease activity, hydro­ D-maltose, potassium gluconate, capric acid, adipic acid, lysis of gelatin, oxidase activity and utilization of L-arabi­ malic acid, trisodium citrate and phenylacetic acid. Strain nose, capric acid, adipic acid and phenylacetic acid. Strain SO98 (=NIBRBAC000503304) was isolated from forest KR3 (=NIBRBAC000503401) was isolated from salted, soil in Yeongju, Gyeongsangbuk Province, Korea (36°51′ fermented scallop (jeotgal) in Anseong, Gyeonggi Prov­ 27.2″N 128°27′28.6″E). ince, Korea (37°0′39.15″N 127°15′50.82″E). Description of Microbacterium nanhaiense LPB0322 Description of Microbacterium invictum 19D1C16 Cells are aerobic, Gram-staining-positive, non-flagel­ Cells are aerobic, Gram-staining-positive, non-flagel­ lated and rod shaped. Colonies are circular, convex, entire lated and rod shaped. Colonies are circular, convex, glis­ and yellow colored after incubation for 3 days on R2A tening and light yellow colored after incubation for 4 days agar medium at 25℃. In the API 20NE system, positive on MH agar at 30℃. In the API 20NE system, positive reaction for esculin hydrolysis, β-galactosidase activity reaction for hydrolysis of esculin and gelatin (weak) and and utilization of potassium gluconate. In the API 20NE β-galactosidase activity. In the API 20NE system, nega­tive system, negative reaction for nitrate reduction, indole pro­ reaction for nitrate reduction, indole production, glucose duction, glucose fermentation, arginine dihydrolase, ure­ fermentation, arginine dihydrolase, urease activity, oxidase ase activity, gelatin hydrolysis, oxidase activity and utili­ activity and utilization of D-glucose, L-arabinose, D-man­ zation of D-glucose, L-arabinose, D-mannose, D-manni­ nose, D-mannitol, N-acetyl-glucosamine, D-maltose, tol, N-acetyl-glucosamine, D-maltose, capric acid, adipic potassium gluconate, capric acid, adipic acid, malic acid, acid, malic acid, trisodium citrate and phenylacetic acid. trisodium citrate and phenylacetic acid. Strain 19D1C16 Strain LPB0322 (=NIBRBAC000503354) was isolated (=NIBRBAC000503259) was isolated from soil around from seashore sand in Taean, Chungcheongnam Province, the Dong River, Gangwon Province, Korea (37°22′56.8″N Korea (36°29′14.5″N 126°20′4.67″E). 128°40′01.2″E). Description of Microbacterium shaanxiense SO111 Description of Microbacterium lemovicicum 19D1A9 Cells are aerobic, Gram-staining-positive, non-flagel­ Cells are aerobic, Gram-staining-positive, non-flagel­ lated and rod shaped. Colonies are circular, smooth, opaque lated and short rod shaped. Colonies are circular, convex, and cream colored after incubation for 3 days on TSA at February 2021 Kim et al. Unrecorded Actinobacteria species of Korea 35

25℃. In the API 20NE system, positive reaction for nit­ agar at 30℃. In the API 20NE system, positive reaction rate reduction, esculin hydrolysis and β-galactosidase for esculin hydrolysis, β-galactosidase activity and utili­ activity. In the API 20NE system, negative reaction for zation of D-glucose (weak), L-arabinose, D-mannose, D- indole production, glucose fermentation, arginine dihydro­ mannitol and D-maltose (weak). In the API 20NE system, lase, urease activity, gelatin hydrolysis, oxidase activity negative reaction for nitrate reduction, indole production, and utilization of D-glucose, L-arabinose, D-mannose, glucose fermentation, arginine dihydrolase, urease activity, D-mannitol, N-acetyl-glucosamine, D-maltose, potassium gelatin hydrolysis, oxidase activity and utilization of N-ace­ gluconate, capric acid, adipic acid, malic acid, trisodium tyl-glucosamine, potassium gluconate, capric acid, adipic citrate and phenylacetic acid. Strain SO111 (=NIBRBAC acid, malic acid, trisodium citrate and phenylacetic acid. 000503305) was isolated from forest soil in Yeongju, Strain 19D2C13 (=NIBRBAC000503269) was isolated Gyeong­sangbuk Province, Korea (36°51′27.2″N 128°27′ from soil around the Dong River, Gangwon Province, Kor­ 28.6″E). ea (37°18′48.5″N 128°37′37.6″E).

Description of Microbacterium thalassium N40 Description of Arthrobacter tumbae 19D2A1 Cells are aerobic, Gram-staining-positive, flagellated Cells are aerobic, Gram-staining-positive, non-flagel­ and rod shaped. Colonies are circular, convex and yellow lated and coccoid. Colonies are circular, convex, smooth colored after incubation for 3 days on R2A agar at 25℃. and cream colored after incubation for 4 days on MH agar In the API 20NE system, positive reaction for nitrate at 30℃. In the API 20NE system, positive reaction for reduction, glucose fermentation, hydrolysis of esculin esculin hydrolysis, β-galactosidase activity and utilization and gelatin and utilization of D-glucose, D-mannose, of D-mannitol and D-maltose. In the API 20NE system, D-mannitol, D-maltose and potassium gluconate. In the negative reaction for nitrate reduction, indole production, API 20NE system, negative reaction for indole produc­ glucose fermentation, arginine dihydrolase, urease activity,­ tion, arginine dihydrolase, urease activity, β-galactosidase gelatin hydrolysis, oxidase activity and utilization of D- activity, oxidase activity and utilization of L-arabinose, glucose, L-arabinose, D-mannose, N-acetyl-glucosamine, N-acetyl-glucosamine, capric acid, adipic acid, malic potassium gluconate, capric acid, adipic acid, malic acid, acid, trisodium citrate and phenylacetic acid. Strain N40 trisodium citrate and phenylacetic acid. Strain 19D2A1 (=NIBRBAC000503417) was isolated from ginseng-cul­ (=NIBRBAC000503267) was isolated from soil around tivated soil in Yeongju, Gyeongsangbuk Province, Korea the Dong River, Gangwon Province, Korea (37°18′48.5″N (36°53′51.5″N 128°28′02.3″E). 128°37′37.6″E).

Description of Microbacterium trichothecenolyticum Description of Paenarthrobacter ilicis 13H-2 BSSP-R25 Cells are aerobic, Gram-staining-positive, non-flagel­ Cells are aerobic, Gram-staining-positive, non-flagella­ lated and coccoid or rod shaped. Colonies are circular and ted and rod shaped. Colonies are circular, slightly convex, yellow colored after incubation for 3 days on R2A agar at glistening and light yellowish pink colored after incuba­ 20-40℃. In the API 20NE system, positive reaction for tion for 3 days on R2A agar at 25℃. In the API 20NE urease activity, hydrolysis of esculin and gelatin, β-galac­ system, positive reaction for hydrolysis of esculin and gel­ tosidase activity and utilization of D-glucose, L-arabinose, atin, β-galactosidase activity and utilization of D-glucose, D-mannose, D-mannitol, N-acetyl-glucosamine, D-mal­ L-arabinose, D-mannose and D-maltose. In the API 20NE tose, potassium gluconate, malic acid, trisodium citrate system, negative reaction for nitrate reduction, indole pro­ and phenylacetic acid. In the API 20NE system, negative duction, glucose fermentation, arginine dihydrolase, ure­ reaction for nitrate reduction, indole production, glucose ase activity, oxidase activity and utilization of D-mannitol, fermentation, arginine dihydrolase, oxidase activity and N-acetyl-glucosamine, potassium gluconate, capric acid, utilization of capric acid and adipic acid. Strain 13H-2 adipic acid, malic acid, trisodium citrate and phenylacetic (=NIBRBAC000503223) was isolated from meadow soil acid. Strain BSSP-R25 (=NIBRBAC000503329) was iso­- in Yeongwol, Gangwon Province, Korea (37°23′04.0″N lated from tidal flat sediment in Boryeong, Chungcheong- 128°41′01.3″E). nam Province, Korea (36°20′15″N 126°53′93″E). Description of Pseudarthrobacter phenanthrenivorans Description of Mycetocola manganoxydans 19D2C13 19D1F19 Cells are aerobic, Gram-staining-positive, flagellated Cells are facultatively aerobic, Gram-staining-positive, and rod shaped. Colonies are circular, convex, glistening non-flagellated and rod shaped. Colonies are circular, and yellow colored after incubation for 4 days on MH raised, glistening and light yellow colored after incubation 36 JOURNAL OF SPECIES RESEARCH Vol. 10, No. 1 for 4 days on MH agar at 30℃. In the API 20NE system, marine agar at 25℃. In the API 20NE system, positive positive reaction for nitrate reduction, esculin hydroly­ reaction for urease activity, hydrolysis of esculin and gela­ sis, β-galactosidase activity and utilization of D-glucose, tin and utilization of D-glucose, D-mannose, D-mannitol D-mannose, D-mannitol, D-maltose, potassium gluconate, and D-maltose. In the API 20NE system, negative reac­ adipic acid, malic acid, trisodium citrate and phenylacetic tion for nitrate reduction, indole production, glucose fer­ acid. In the API 20NE system, negative reaction for indole mentation, arginine dihydrolase, β-galactosidase activity, production, glucose fermentation, arginine dihydrolase, oxidase activity and utilization of L-arabinose, N-acetyl- urease activity, gelatin hydrolysis, oxidase activity and uti­ glucosamine, potassium gluconate, capric acid, adipic acid, lization of L-arabinose, N-acetyl-glucosamine and capric malic acid, trisodium citrate and phenylacetic acid. Strain acid. Strain 19D1F19 (=NIBRBAC000503260) was iso­ KYW1971 (=NIBRBAC000503307) was isolated from lated from soil around the Dong River, Gangwon Prov­ seawater in Gwangyang, Jeollanam Province, Korea (34° ince, Korea (37°22′56.8″N 128°40′01.2″E). 54′24.83″N 127°44′01.47″E).

Description of Pseudarthrobacter polychromogenes Description of Longispora urticae R21 JBTF-M16 Cells are aerobic, Gram-staining-positive, non-flagel­ Cells are aerobic, Gram-staining-negative, non-flagella­ lated and rod shaped. Colonies are circular and yellowish ted and rod shaped. Colonies are circular, convex, glisten­ white colored after incubation for 3 days on R2A agar at ing and yellowish white colored after incubation for 2 days 10-37℃. In the API 20NE system, positive reaction for on marine agar medium at 30℃. In the API 20NE sys­ nitrate reduction, hydrolysis of gelatin, β-galactosidase tem, positive reaction for nitrate reduction, hydrolysis of acti­vity, oxidase activity and utilization of potassium gluco­ esculin and gelatin, β-galactosidase activity and utilization nate. In the API 20NE system, negative reaction for indole of D-glucose, D-mannose, D-mannitol, N-acetyl-glucosa­ production, glucose fermentation, arginine dihydro­lase, mine, D-maltose, potassium gluconate, malic acid, triso­ urease activity, hydrolysis of esculin and utilization of D- dium citrate and phenylacetic acid. In the API 20NE sys­ glucose, L-arabinose, D-mannose, D-mannitol, N-acetyl- tem, negative reaction for indole production, glucose fer­ glucosamine, D-maltose, capric acid, adipic acid, malic mentation, arginine dihydrolase, urease activity, oxidase acid, trisodium citrate and phenylacetic acid. Strain R21 activity and utilization of L-arabinose, capric acid and adi­ ( =NIBRBAC000503218) was isolated from a soil in pic acid. Strain JBTF-M16 (=NIBRBAC000503335) was Yeongwol, Gangwon Province, Korea (37°15′38.4″N 128° isolated from tidal flat sediment in Jebu Island, Gyeonggi 36′28.7″E). Province, Korea (37°9′48″N 126°37′1″E). Description of Micromonospora avicenniae R_77 Description of LPB0310 Cells are aerobic, Gram-staining-positive, non-flagella­ Cells are aerobic, Gram-staining-positive, non-flagel­ ted and filamentous. Colonies are circular, convex, rough, lated and rod shaped. Colonies are circular, rhizoid, um­ entire and orange colored after incubation for 3 days on bonate and white colored after incubation for 3 days on ISP7 agar at 30℃. In the API 20NE system, positive re­ blood agar at 37℃. In the API 20NE system, positive action for nitrate reduction, hydrolysis of esculin and gel­ reaction for nitrate reduction, hydrolysis (weak) of esculin atin, β-galactosidase activity and utilization of D-glucose, and gelatin and oxidase activity. In the API 20NE sys­ L-arabinose, D-mannose, D-mannitol, D-maltose, potas­ tem, negative reaction for indole production, glucose sium gluconate, adipic acid, malic acid and phenylacetic fermentation, arginine dihydrolase, urease activity, β-gal­ acid. In the API 20NE system, negative reaction for indole actosidase activity and utilization of D-glucose, L-arabi­ production, glucose fermentation, arginine dihydrolase, nose, D-mannose, D-mannitol, N-acetyl-glucosamine, D- urease activity, oxidase activity and utilization of N-ace­ maltose, potassium gluconate, capric acid, adipic acid, tyl-glucosamine, capric acid and trisodium citrate. Strain malic acid, trisodium citrate and phenylacetic acid. Strain R_77 (=NIBRBAC000503392) was isolated from soil in LPB0310 (=NIBRBAC000503349) was isolated from a Sejong, Korea (36°28′42.4″N 127°15′42.0″E). human respiratory organ in Seoul, Korea (37°34′47.74″N 126°59′56.12″E). Description of Micromonospora oryzae G92 Cells are aerobic, Gram-staining-positive and coccoid. Description of Rothia amarae KYW1971 Colonies are filamentous, umbonate and orange colored Cells are aerobic, Gram-staining-positive, non-flagella­ after incubation for 3 days on R2A agar at 25℃. In the ted and coccoid. Colonies are circular, convex, smooth, API 20NE system, positive reaction for arginine dihydro- opaque and white colored after incubation for 2 days on lase, urease activity, hydrolysis of esculin and gelatin and February 2021 Kim et al. Unrecorded Actinobacteria species of Korea 37 utilization of D-glucose, L-arabinose, D-maltose and nate, malic acid and trisodium citrate. In the API 20NE potassium gluconate. In the API 20NE system, negative system, negative reaction for indole production, arginine reaction for nitrate reduction, indole production, glucose dihydrolase, hydrolysis of esculin and gelatin, β-galactosi­ fermentation, β-galactosidase activity, oxidase activity dase activity, oxidase activity and utilization of D-glucose, and utilization of D-mannose, D-mannitol, N-acetyl-glu­ L-arabinose, D-mannose, N-acetyl-glucosamine, D-mal­ cosamine, capric acid, adipic acid, malic acid, trisodium tose, capric acid, adipic acid and phenylacetic acid. Strain citrate and phenylacetic acid. Strain G92 (=NIBRBAC FS100 (=NIBRBAC000503294) was isolated from soil 000503406) was isolated from ginseng-cultivated soil in in Suncheon, Jeollanam Province, Korea (34°58′12.4″N Yeongju, Gyeongsangbuk Province, Korea (36°53′51.5″N 127°28′53.0″E). 128°28′02.3″E). Description of Mycolicibacterium wolinskyi S5 Description of Micromonospora schwarzwaldensis Cells are aerobic, Gram-staining-positive, non-flagel­ BT360 lated and rod shaped. Colonies are circular, convex and Cells are aerobic, Gram-staining-positive, non-flagel­ cream colored after incubation for 3 days on R2A agar at lated and rod shaped. Colonies are circular, smooth and 25℃. In the API 20NE system, positive reaction for oxi­ orange colored after incubation for 3 days on R2A agar at dase activity and utilization of D-glucose, D-mannose, D- 25℃. In the API 20NE system, positive reaction for nit­ mannitol, potassium gluconate and malic acid. In the API rate reduction, hydrolysis of gelatin (weak), β-galactosi­ 20NE system, negative reaction for nitrate reduction, in­ dase activity (weak) and oxidase activity. In the API 20NE dole production, glucose fermentation, arginine dihydro­ system, negative reaction for indole production, glucose lase, urease activity, hydrolysis of esculin and gelatin, β- fermentation, arginine dihydrolase, urease activity, hydro­ galactosidase activity and utilization of L-arabinose, N- lysis of esculin and utilization of D-glucose, L-arabinose, acetyl-glucosamine, D-maltose, capric acid, adipic acid, D-mannose, D-mannitol, N-acetyl-glucosamine, D-mal­ trisodium citrate and phenylacetic acid. Strain S5 (=NIBR tose, potassium gluconate, capric acid, adipic acid, malic BAC000503408) was isolated from forest soil in Yeongju, acid, trisodium citrate and phenylacetic acid. Strain BT360 Gyeongsangbuk Province, Korea (36°54′12.67″N 128°27′ (=NIBRBAC000503002) was isolated from soil in Jeju 32.36″E). Island, Korea (33°28′58.6″N 126°30′44.6″E). Description of Nocardia alba 19D2V10 Description of Dietzia lutea SR3 Cells are facultatively aerobic, Gram-staining-positive, Cells are aerobic, Gram-staining-positive, non-flagel­ non-flagellated and rod shaped. Colonies are circular, flat lated and coccoid. Colonies are circular, convex and light and white colored after incubation for 4 days on MH agar orange colored after incubation for 3 days on R2A agar at at 30℃. In the API 20NE system, positive reaction for 25℃. In the API 20NE system, positive reaction for utili­ nitrate reduction, urease activity, hydrolysis of esculin, zation of D-glucose, D-mannose, potassium gluconate and β-galactosidase activity and utilization of D-glucose, adipic acid. In the API 20NE system, negative reaction L-arabinose, D-mannose, D-mannitol, N-acetyl-glucosa­ for nitrate reduction, indole production, glucose fermen­ mine, potassium gluconate, adipic acid and malic acid. tation, arginine dihydrolase, urease activity, hydrolysis of In the API 20NE system, negative reaction for indole esculin and gelatin, β-galactosidase activity, oxidase acti­ production, glucose fermentation, arginine dihydrolase, vity and utilization of L-arabinose, D-mannitol, N-acetyl- hydrolysis of gelatin, oxidase activity and utilization of glucosamine, D-maltose, capric acid, malic acid, triso­ D-maltose, capric acid, trisodium citrate and phenylacetic dium citrate and phenylacetic acid. Strain SR3 (=NIBR acid. Strain 19D2V10 ( =NIBRBAC000503270) was BAC000503402) was isolated from salted, fermented isolated from soil in the Dong River, Gangwon Province, shrimp (jeotgal) in Anseong, Gyeonggi Province, Korea Korea (37°18′48.5″N 128°37′37.6″E). (37°0′39.15″N 127°15′50.82″E). Description of Nocardia grenadensis 19D1S1 Description of Williamsia limnetica FS100 Cells are facultatively aerobic, Gram-staining-positive, Cells are aerobic, Gram-staining-positive, non-flagella­ non-flagellated and rod shaped. Colonies are circular, um­ ted and rod shaped. Colonies are circular, convex, smooth, bonate and white colored after incubation for 4 days on opaque and pale pink colored after incubation for 3 days MH agar at 30℃. In the API 20NE system, positive reac­ on R2A agar at 25℃. In the API 20NE system, positive tion for hydrolysis of esculin and utilization of D-glucose, reaction for nitrate reduction, glucose fermentation, urease potassium gluconate, adipic acid and malic acid. In the activity and utilization of D-mannitol, potassium gluco­ API 20NE system, negative reaction for nitrate reduction, 38 JOURNAL OF SPECIES RESEARCH Vol. 10, No. 1 indole production, glucose fermentation, arginine dihydro­ of esculin, β-galactosidase activity, oxidase activity and lase, urease activity, hydrolysis of gelatin, β-galactosidase utilization of D-mannitol, N-acetyl-glucosamine, D-mal­ activity, oxidase activity and utilization of L-arabinose, tose, potassium gluconate, capric acid, adipic acid, malic D-mannose, D-mannitol, N-acetyl-glucosamine, D-mal­ acid, trisodium citrate and phenylacetic acid. Strain R12 tose, capric acid, trisodium citrate and phenylacetic acid. (=NIBRBAC000503418) was isolated from ginseng-cul­ Strain 19D1S1 ( =NIBRBAC000503264) was isolated tivated soil in Yeongju, Gyeongsangbuk Province, Korea from soil in the Dong River, Gangwon Province, Korea (36°53′51.5″N 128°28′02.3″E). (37°22′56.8″N 128°40′01.2″E). Description of BSSP-M28 Description of Nocardia nova 19D1V24 Cells are aerobic, Gram-staining-positive, non-flagel­ Cells are facultatively aerobic, Gram-staining-positive, lated and rod shaped. Colonies are circular, slightly con­ non-flagellated and rod shaped. Colonies are circular, vex, glistening and yellowish white colored after incuba­ umbonate and white colored after incubation for 4 days tion for 5 days on marine agar at 25℃. In the API 20NE on MH agar at 30℃. In the API 20NE system, positive system, positive reaction for hydrolysis of gelatin and uti­ reaction for hydrolysis of esculin and utilization of adipic lization of D-glucose. In the API 20NE system, negative acid. In the API 20NE system, negative reaction for nit­ reaction for nitrate reduction, indole production, glucose rate reduction, indole production, glucose fermentation, fermentation, arginine dihydrolase, urease activity, hydro­ arginine dihydrolase, urease activity, hydrolysis of gelatin, lysis of esculin, β-galactosidase activity, oxidase activity β-galactosidase activity, oxidase activity and utilization of and utilization of L-arabinose, D-mannose, D-mannitol, D-glucose, L-arabinose, D-mannose, D-mannitol, N-ace­ N-acetyl-glucosamine, D-maltose, potassium gluconate, tyl-glucosamine, D-maltose, potassium gluconate, capric capric acid, adipic acid, malic acid, trisodium citrate and acid, malic acid, trisodium citrate and phenylacetic acid. phenylacetic acid. Strain BSSP-M28 (=NIBRBAC00050 Strain 19D1V24 (=NIBRBAC000503272) was isolated 3336) was isolated from tidal flat sediment in Boryeong, from soil in the Dong River, Gangwon Province, Korea Chungcheongnam Province, Korea (36°20′15″N 126°53′ (37°22′56.8″N 128°40′01.2″E). 93″E).

Description of Nocardia tengchongensis JDB110 Description of Marmoricola aquaticus 19D1C14 Cells are aerobic, Gram-staining-positive, non-flagel­ Cells are aerobic, Gram-staining-positive, non-flagel­ lated and rod shaped. Colonies are circular, flat, smooth, lated and pleomorphic. Colonies are circular, convex and opaque and white colored after incubation for 4 days on orange colored after incubation for 4 days on MH agar at TSA at 25℃. In the API 20NE system, positive reaction 30℃. In the API 20NE system, positive reaction for hydro­ for nitrate reduction, hydrolysis of esculin and utilization lysis of esculin and gelatin, β-galactosidase activity and of N-acetyl-glucosamine and malic acid. In the API 20NE utilization of D-glucose, L-arabinose, D-mannose (weak), system, negative reaction for indole production, glucose D-mannitol, N-acetyl-glucosamine, D-maltose, potassium fermentation, arginine dihydrolase, urease activity, hydro­ gluconate, adipic acid (weak) and malic acid. In the API lysis of gelatin, β-galactosidase activity, oxidase activity 20NE system, negative reaction for nitrate reduction, indole and utilization of D-glucose, L-arabinose, D-mannose, production, glucose fermentation, arginine dihydrolase,­ ure­ D-mannitol, D-maltose, potassium gluconate, capric acid, ase activity, oxidase activity and utilization of capric acid, adipic acid, trisodium citrate and phenylacetic acid. Strain trisodium citrate and phenylacetic acid. Strain 19D1C14 JDB110 (=NIBRBAC000503295) was isolated from soil (=NIBRBAC000503258) was isolated from soil in the in Goheung, Jeollanam Province, Korea (34°27′29.29″N Dong River, Gangwon Province, Korea (37°22′56.8″N 127°11′14.21″E). 128°40′01.2″E).

Description of Rhodococcus artemisiae R12 Description of Nocardioides phosphati BT343 Cells are aerobic, Gram-staining-positive, non-flagella­ Cells are aerobic, Gram-staining-positive, non-flagella­ ted and rod shaped. Colonies are smooth, convex and pink ted and rod shaped. Colonies are circular, convex, smooth colored after incubation for 3 days on R2A agar at 25℃. and white colored after incubation for 3 days on R2A agar In the API 20NE system, positive reaction for indole pro­ at 25℃. In the API 20NE system, positive reaction for duction, urease activity, hydrolysis of gelatin and utili­ nitrate reduction, arginine dihydrolase, urease activity and zation of D-glucose, L-arabinose and D-mannose. In the utilization of D-glucose, L-arabinose (weak), N-acetyl-glu­ API 20NE system, negative reaction for nitrate reduction, cosamine (weak), potassium gluconate, capric acid (weak), glucose fermentation, arginine dihydrolase, hydrolysis adipic acid (weak), malic acid and trisodium citrate. In the February 2021 Kim et al. Unrecorded Actinobacteria species of Korea 39

API 20NE system, negative reaction for indole production, production, glucose fermentation, arginine dihydrolase, glucose fermentation, hydrolysis of esculin and gelatin, urease activity, hydrolysis of esculin, and utilization of β-galactosidase activity, oxidase activity and utilization of L-arabinose, D-mannose, D-mannitol, D-maltose, capric D-mannose, D-mannitol, D-maltose and phenylacetic acid. acid, adipic acid, trisodium citrate and phenylacetic acid. Strain BT343 ( =NIBRBAC000502998) was isolated Strain MMS19-T35 (=NIBRBAC000503382) was isola­ from soil in Jeju Island, Korea (33°28′30.5″N 126°30′ ted from soil in Gurye, Jeollanam Province, Korea (35°16′ 39.4″E). 25.9″N 127°28′34.5″E).

Description of Lentzea albidocapillata subsp. Description of Kitasatospora xanthocidica LPB0280 albidocapillata BSSP-M29 Cells are aerobic, Gram-staining-positive, non-flagel­ Cells are aerobic, Gram-staining-positive, non-flagel­ lated and rod shaped. Colonies are irregular, undulate, flat lated and rod shaped. Colonies are circular and yellowish and cream colored after incubation for 3 days on TSA at gray colored after incubation for 3 days on marine agar at 25℃. In the API 20NE system, positive reaction for hydro­ 30℃. In the API 20NE system, positive reaction for argi­ lysis of esculin (weak), oxidase activity and utilization of nine dihydrolase, urease activity, hydrolysis of esculin and D-glucose, L-arabinose (weak), N-acetyl-glucosamine, gelatin, β-galactosidase activity and utilization of D-glu­ potassium gluconate, malic acid (weak) and trisodium cit­ cose, L-arabinose, D-mannose, D-mannitol, N-acetyl-glu­ rate (weak). In the API 20NE system, negative reaction for cosamine, D-maltose and malic acid. In the API 20NE nitrate reduction, indole production, glucose fermentation, system, negative reaction for nitrate reduction, indole arginine dihydrolase, urease activity, hydrolysis of gela­ production, glucose fermentation, oxidase activity and uti­ tin, β-galactosidase activity and utilization of D-mannose, lization of potassium gluconate, capric acid, adipic acid, D-mannitol, D-maltose, capric acid, adipic acid and phe­ trisodium citrate and phenylacetic acid. Strain BSSP-M29 nylacetic acid. Strain LPB0280 (=NIBRBAC000503341) (=NIBRBAC000503328) was isolated from tidal flat sedi-­ was isolated from soil in Seoul, Korea (37°35′5.01″N 127° ment in Boryeong, Chungcheongnam Province, Korea 1′35.69″E). (36°20′15″N 126°53′93″E). Description of Streptacidiphilus carbonis S36 Description of Lentzea guizhouensis BT46 Cells are aerobic, Gram-staining-positive and filamen­ Cells are aerobic, Gram-staining-positive, non-flagella­ tous. Colonies are filamentous, umbonate and white col­ ted and rod shaped. Colonies are circular, convex, smooth ored after incubation for 3 days on R2A agar at 25℃. In and white colored after incubation for 3 days on R2A agar the API 20NE system, positive reaction for hydrolysis at 25℃. In the API 20NE system, positive reaction for of esculin and utilization of D-glucose, D-mannitol, N- nitrate reduction, hydrolysis (weak) of esculin and gela­ acetyl-glucosamine and potassium gluconate. In the API tin and β-galactosidase activity (weak). In the API 20NE 20NE system, negative reaction for nitrate reduction, in­ system, negative reaction for indole production, glucose dole production, glucose fermentation, arginine dihydro- fermentation, arginine dihydrolase, urease activity, oxi­ lase, urease activity, hydrolysis of gelatin, β-galactosi­ dase activity and utilization of D-glucose, L-arabinose, D- dase activity, oxidase activity and utilization of L-arabi­ mannose, D-mannitol, N-acetyl-glucosamine, D-maltose, nose, D-mannose, D-maltose, capric acid, adipic acid, potassium gluconate, capric acid, adipic acid, malic acid, tri­ malic acid, trisodium citrate and phenylacetic acid. Strain sodium citrate and phenylacetic acid. Strain BT46 (=NIBR S36 (=NIBRBAC000503409) was isolated from forest BAC000502986) was isolated from soil in Pyeongchang, soil in Yeongju, Gyeongsangbuk Province, Korea (36°54′ Gangwon Province, Korea (37°42′21.1″N 128°43′01.9″E). 12.67″N 128°27′32.36″E).

Description of Kitasatospora purpeofusca Description of Streptomyces albogriseolus R-5 MMS19-T35 Cells are aerobic, Gram-staining-positive, non-flagella­ Cells are aerobic, Gram-staining-positive, non-flagel­ ted and rod shaped. Colonies are filamentous, raised, undu­ lated and filamentous or rod shaped. Colonies are circu­ late and cream colored after incubation for 3 days on R2A lar, convex, erose and brown colored after incubation for agar at 37℃. In the API 20NE system, positive reaction 3 days on R2A agar at 30℃. In the API 20NE system, for nitrate reduction, glucose fermentation (weak), hydro­ positive reaction for nitrate reduction, hydrolysis of gela­ lysis of esculin (weak) and gelatin, β-galactosidase activi­ tin, β-galactosidase activity and utilization of D-glucose, ty and utilization of D-glucose, L-arabinose, D-mannose, N-acetyl-glucosamine, potassium gluconate and malic D-mannitol, N-acetyl-glucosamine, D-maltose, potassium acid. In the API 20NE system, negative reaction for indole gluconate, adipic acid (weak), malic acid and trisodium 40 JOURNAL OF SPECIES RESEARCH Vol. 10, No. 1 citrate. In the API 20NE system, negative reaction for in­ gluconate, capric acid, adipic acid, malic acid, trisodium dole production, arginine dihydrolase, urease activity and citrate and phenylacetic acid. Strain 9C-1 (=NIBRBAC utilization of capric acid and phenylacetic acid. Strain R-5 000503221) was isolated from soil in Yeongwol, Gang­ (=NIBRBAC000503388) was isolated from soil in Gu­- won Province, Korea (37°13′52.7″N 128°28′59.0″E). rye, Jeollanam Province, Korea (35°11′16.0″N 127°33′ 02.5″E). Description of Streptomyces brevispora SO100 Cells are aerobic, Gram-staining-positive, non-flagella­ Description of Streptomyces amakusaensis 19D2C16 ted and filamentous. Colonies are circular, smooth, opaque Cells are aerobic, Gram-staining-positive, non-flagel­ and white to gray colored after incubation for 4 days lated and filamentous. Colonies are circular, raised and on R2A agar at 25℃. In the API 20NE system, positive brown colored after incubation for 4 days on MH agar at reaction for nitrate reduction, urease activity, hydrolysis 30℃. In the API 20NE system, positive reaction for hydro- of esculin and gelatin, β-galactosidase activity and utiliza­ lysis of esculin and gelatin, oxidase activity and utilization tion of D-glucose, L-arabinose, D-mannose, N-acetyl-glu­ of D-glucose, N-acetyl-glucosamine, potassium gluco­ cosamine, D-maltose and potassium gluconate. In the API nate, adipic acid and malic acid. In the API 20NE system, 20NE system, negative reaction for indole production, negative reaction for nitrate reduction, indole production, glucose fermentation, arginine dihydrolase, oxidase acti­ glucose fermentation, arginine dihydrolase, urease acti­ vity and utilization of D-mannitol, capric acid, adipic acid, vity, β-galactosidase activity and utilization of L-arabi­ malic acid, trisodium citrate and phenylacetic acid. Strain nose, D-mannose, D-mannitol, D-maltose, capric acid, SO100 (=NIBRBAC000503306) was isolated from soil trisodium citrate and phenylacetic acid. Strain 19D2C16 in Yeongju, Gyeongsangbuk Province, Korea (36°51′27.2″ (=NIBRBAC000503276) was isolated from soil around N 128°27′28.6″E). the Dong River, Gangwon Province, Korea (37°18′48.5″N 128°37′37.6″E). Description of subsp. asoensis BG138 Description of BT63 Cells are aerobic, Gram-staining-positive, non-flagella­ Cells are aerobic, Gram-staining-positive, non-flagella­ ted and filamentous. Colonies are circular, convex, entire ted and rod shaped. Colonies are circular, smooth and yel­ and cream colored after incubation for 3 days on TSA at low colored after incubation for 3 days on 1/10 LB agar at 30℃. In the API 20NE system, positive reaction for nitrate 25℃. In the API 20NE system, positive reaction for nitrate reduction, glucose fermentation (weak), arginine dihydro­ reduction, urease activity, hydrolysis of gelatin (weak), lase (weak), urease activity (weak), hydrolysis of esculin β-galactosidase activity (weak) and oxidase activity.­ and gelatin, β-galactosidase activity, oxidase activity and In the API 20NE system, negative reaction for indole utilization of D-glucose, L-arabinose, D-mannose, D-man­ production, glucose fermentation, arginine dihydrolase, nitol, N-acetyl-glucosamine, D-maltose (weak), potassium hydrolysis of esculin and utilization of D-glucose, L-arabi­ gluconate, adipic acid (weak), malic acid (weak) and tri­ nose, D-mannose, D-mannitol, N-acetyl-glucosamine, sodium citrate (weak). In the API 20NE system, negative D-mal­tose, potassium gluconate, capric acid, adipic acid, reaction for indole production and utilization of capric malic acid, trisodium citrate and phenylacetic acid. Strain acid and phenylacetic acid. Strain BG138 (=NIBRBAC BT63 (=NIBRBAC000502992) was isolated from soil in 000503380) was isolated from soil in Gurye, Jeollanam Pyeongchang, Gangwon Province, Korea (37°42′04.5″N Province, Korea (35°16′25.9″N 127°28′34.5″E). 128°42′49.1″E). Description of EAC34 Description of 9C-1 Cells are aerobic, Gram-staining-positive, non-flagel­ Cells are aerobic, Gram-staining-positive, non-flagella­ lated and rod shaped. Colonies are circular, umbonate, ted and filamentous. Colonies are circular, spore-forming opaque and creamy white colored after incubation for 3 and yellow colored after incubation for 3 days on R2A agar days on R2A agar at 25℃. In the API 20NE system, posi­ at 20-40℃. In the API 20NE system, positive reaction tive reaction for nitrate reduction, indole production, ure­ for urease activity, hydrolysis of esculin and gelatin and ase activity, hydrolysis of esculin and gelatin, β-galacto­ β-galactosidase activity. In the API 20NE system, nega­ sidase activity and utilization of D-glucose, N-acetyl-glu­ tive reaction for nitrate reduction, indole production, glu­ cosamine, potassium gluconate, malic acid and trisodium cose fermentation, arginine dihydrolase, oxidase activity­ citrate. In the API 20NE system, negative reaction for glu­ and utilization of D-glucose, L-arabinose, D-mannose, cose fermentation, arginine dihydrolase, oxidase activity D-mannitol, N-acetyl-glucosamine, D-maltose, potassium and utilization of L-arabinose, D-mannose, D-mannitol, February 2021 Kim et al. Unrecorded Actinobacteria species of Korea 41

D-maltose, capric acid, adipic acid and phenylacetic acid. acid, trisodium citrate and phenylacetic acid. Strain MMS Strain EAC34 ( =NIBRBAC000503282) was isolated 19-T27 (=NIBRBAC000503371) was isolated from soil from soil in Jeju Island, Korea (33°24′33.6″N 126°20′ in Gurye, Jeollanam Province, Korea (35°16′25.9″N 127° 26.3″E). 28′34.5″E).

Description of Streptomyces coelescens 19D1L39 Description of Streptomyces finlayi 5C-2 Cells are facultatively aerobic, Gram-staining-positive, Cells are aerobic, Gram-staining-positive, non-flagel­ non-flagellated and filamentous. Colonies are circular, con­ lated and filamentous. Colonies are circular, spore-form­ vex and orange colored after incubation for 4 days on MH ing and gray white colored after incubation for 3 days on agar at 30℃. In the API 20NE system, positive reaction­ R2A agar at 20-40℃. In the API 20NE system, positive for nitrate reduction, urease activity, hydrolysis of esculin reaction for hydrolysis of esculin and gelatin, β-galacto­ and gelatin, β-galactosidase activity and utilization of sidase activity and utilization of D-glucose, D-mannitol, D-glucose, L-arabinose, D-mannose, D-mannitol, N-ace­ D-maltose, potassium gluconate and malic acid. In the tyl-glucosamine, D-maltose, potassium gluconate, adipic API 20NE system, negative reaction for nitrate reduc­ acid, malic acid, trisodium citrate and phenylacetic acid tion, indole production, glucose fermentation, arginine (weak). In the API 20NE system, negative reaction for in­ dihydrolase, urease activity, oxidase activity and utiliza­ dole production, glucose fermentation, arginine dihydro­ tion of L-arabinose, D-mannose, N-acetyl-glucosamine, lase, oxidase activity and utilization of capric acid. Strain capric acid, adipic acid, trisodium citrate and phenylacetic 19D1L39 ( =NIBRBAC000503274) was isolated from acid. Strain 5C-2 (=NIBRBAC000503220) was isolated soil around the Dong River, Gangwon Province, Korea from cave soil in Yeongwol, Gangwon Province, Korea (37°22′56.8″N 128°40′01.2″E). (37°07′46.2″N 128°31′58.6″E).

Description of Streptomyces corchorusii R-9 Description of 5C-1 Cells are aerobic, Gram-staining-positive, non-flagella­ Cells are aerobic, Gram-staining-positive, non-flagella­ ted and rod shaped. Colonies are filamentous, raised, undu­ ted and filamentous. Colonies are circular, spore-forming late and light beige colored after incubation for 3 days on and brown white colored after incubation for 3 days on R2A agar at 30℃. In the API 20NE system, positive reac­ R2A agar at 20-40℃. In the API 20NE system, positive tion for nitrate reduction, hydrolysis of esculin and gelatin, reaction for hydrolysis of esculin and gelatin, β-galacto­ β-galactosidase activity and utilization of D-glucose, sidase activity and utilization of D-glucose, D-mannose, L-arabinose, D-mannose (weak), D-mannitol, D-maltose D-mannitol, N-acetyl-glucosamine, D-maltose, potassium (weak), potassium gluconate and malic acid. In the API gluconate, malic acid and trisodium citrate. In the API 20NE system, negative reaction for indole production, 20NE system, negative reaction for nitrate reduction, in­ glucose fermentation, arginine dihydrolase, urease activity dole production, glucose fermentation, arginine dihydro­ and utilization of N-acetyl-glucosamine, capric acid, adipic lase, urease activity, oxidase activity and utilization of L- acid, trisodium citrate and phenylacetic acid. Strain R9 arabinose, capric acid, adipic acid and phenylacetic acid. (=NIBRBAC000503387) was isolated from soil in Gu- Strain 5C-1 (=NIBRBAC000503219) was isolated from rye, Jeollanam Province, Korea (35°11′16.0″N 127°33′ cave soil in Yeongwol, Gangwon Province, Korea (37°07′ 02.5″E). 46.2″N 128°31′58.6″E).

Description of Streptomyces europaeiscabiei Description of 13H-1 MMS19-T27 Cells are aerobic, Gram-staining-positive, non-flagella­ Cells are aerobic, Gram-staining-positive, non-flagella­ ted and filamentous. Colonies are circular, spore-forming ted and filamentous. Colonies are circular, convex, erose and yellow colored after incubation for 3 days on R2A agar and brown colored after incubation for 3 days on R2A agar at 20-40℃. In the API 20NE system, positive reaction at 30℃. In the API 20NE system, positive reaction for for hydrolysis of esculin and gelatin, β-galactosidase acti­ nitrate reduction, urease activity (weak), hydrolysis of vity and utilization of D-glucose, L-arabinose, D-maltose esculin and gelatin, β-galactosidase activity, oxidase acti­ and potassium gluconate. In the API 20NE system, nega­ vity and utilization of D-glucose, L-arabinose, D-mannose, tive reaction for nitrate reduction, indole production, glu­ D-mannitol, N-acetyl-glucosamine, D-maltose, potassium cose fermentation, arginine dihydrolase, urease activity, gluconate, adipic acid and malic acid. In the API 20NE oxidase activity and utilization of D-mannose, D-mannitol, system, negative reaction for indole production, glucose N-acetyl-glucosamine, capric acid, adipic acid, malic fermentation, arginine dihydrolase and utilization of capric acid, trisodium citrate and phenylacetic acid. Strain 13H-1 42 JOURNAL OF SPECIES RESEARCH Vol. 10, No. 1

(=NIBRBAC000503222) was isolated from meadow soil Description of Streptomyces hydrogenans SO94 in Yeongwol, Gangwon Province, Korea (37°23′04.0″N Cells are aerobic, Gram-staining-positive, non-flagella­ 128°41′01.3″E). ted and filamentous. Colonies are circular, smooth, opaque Description of Streptomyces fulvissimus 19D2F17 and pale yellow colored after incubation for 3 days on R2A agar at 25℃. In the API 20NE system, positive reaction Cells are facultatively aerobic, Gram-staining-positive, for nitrate reduction, indole production, urease activity, non-flagellated and filamentous. Colonies are circular, hydrolysis of esculin and gelatin and utilization of D-glu­ raised and yellow colored after incubation for 4 days on cose, L-arabinose, D-mannose, D-mannitol, potassium MH agar at 30℃. In the API 20NE system, positive reac­ gluconate, adipic acid, malic acid, trisodium citrate and tion for nitrate reduction, urease activity, hydrolysis of phenylacetic acid. In the API 20NE system, negative reac­ esculin and gelatin, β-galactosidase activity and utilization tion for glucose fermentation, arginine dihydrolase, β- of D-glucose, L-arabinose (weak), D-mannose, D-manni­ galactosidase activity, oxidase activity and utilization of tol, N-acetyl-glucosamine, D-maltose (weak), potassium N-acetyl-glucosamine, D-maltose and capric acid. Strain gluconate, adipic acid (weak), malic acid and trisodium SO94 (=NIBRBAC000503297) was isolated from soil in citrate (weak). In the API 20NE system, negative reaction Yeongju, Gyeongsangbuk Province, Korea (36°51′27.2″N for indole production, glucose fermentation, arginine di­ 128°27′28.6″E). hydrolase, oxidase activity and utilization of capric acid = and phenylacetic acid. Strain 19D2F17 ( NIBRBAC Description of 19D2S3 000503277) was isolated from soil around the Dong River, Gangwon Province, Korea (37°18′48.5″N 128°37′37.6″E). Cells are facultatively aerobic, Gram-staining-positive, non-flagellated and filamentous. Colonies are irregular, Description of CAU 1564 flat and brown colored after incubation for 4 days on MH Cells are aerobic, Gram-staining-positive, non-flagella­ agar at 30℃. In the API 20NE system, positive reaction ted and rod shaped. Colonies are circular, smooth, convex, for urease activity, hydrolysis of esculin and gelatin, β-gal- shiny and cream colored after incubation for 3 days on actosidase activity, oxidase activity and utilization of D- nutrient agar at 37℃. In the API 20NE system, positive glucose, D-mannose, N-acetyl-glucosamine, potassium gluconate, adipic acid (weak), malic acid, trisodium citrate reaction for utilization of L-arabinose, D-mannose, N-ace­ tyl-glucosamine (weak) and D-maltose. In the API 20NE and phenylacetic acid (weak). In the API 20NE system, system, negative reaction for nitrate reduction, indole negative reaction for nitrate reduction, indole production, production, glucose fermentation, arginine dihydrolase, glucose fermentation, arginine dihydrolase and utilization urease activity, hydrolysis of esculin and gelatin, β-galac­ of L-arabinose, D-mannitol, D-maltose and capric acid. = tosidase activity and utilization of D-glucose, D-mannitol, Strain 19D2S3 ( NIBRBAC000503278) was isolated from soil around the Dong River, Gangwon Province, potassium gluconate, capric acid, adipic acid, malic acid, ′ ″ ′ ″ trisodium citrate and phenylacetic acid. Strain CAU 1564 Korea­ (37°18 48.5 N 128°37 37.6 E). ( =NIBRBAC000503235) was isolated from seashore sand in Incheon, Korea (37°31′50.8″N 126°25′53.1″E). Description of Streptomyces nigrescens JDB244 Cells are aerobic, Gram-staining-positive, non-flagella­ Description of EAC30 Streptomyces griseorubiginosus ted and filamentous. Colonies are wavy, wrinkled, opaque Cells are aerobic, Gram-staining-positive, non-flagel­ and white to gray colored after incubation for 5 days on lated and filamentous. Colonies are wrinkled, wavy, hilly, R2A agar at 25℃. In the API 20NE system, positive reac­ opaque and brown colored after incubation for 7 days on tion for hydrolysis of gelatin. In the API 20NE system, TSA at 25℃. In the API 20NE system, positive reaction negative reaction for nitrate reduction, indole production, for hydrolysis of esculin, β-galactosidase activity and oxi­ glucose fermentation, arginine dihydrolase, urease activity, dase activity. In the API 20NE system, negative reaction hydrolysis of esculin, β-galactosidase activity, oxidase for nitrate reduction, indole production, glucose fermen­ activity and utilization of D-glucose, L-arabinose, D-man­ tation, arginine dihydrolase, urease activity, hydrolysis of nose, D-mannitol, N-acetyl-glucosamine, D-maltose, gelatin and utilization of D-glucose, L-arabinose, D-man­ potassium gluconate, capric acid, adipic acid, malic acid, nose, D-mannitol, N-acetyl-glucosamine, D-maltose, potas­- trisodium citrate and phenylacetic acid. Strain JDB244 sium gluconate, capric acid, adipic acid, malic acid, triso­ (=NIBRBAC000503303) was isolated from soil in Go­ dium citrate and phenylacetic acid. Strain EAC30 (=NIBR heung, Jeollanam Province, Korea (34°27′29.33″N 127° BAC000503283) was isolated from soil in Jeju Island, 11′14.21″E). Korea (33°24′33.6″N 126°20′26.3″E). February 2021 Kim et al. Unrecorded Actinobacteria species of Korea 43

Description of Streptomyces phaeoluteichromatogenes ted and rod shaped. Colonies are circular, convex, filamen­ R-21 tous and light yellow colored after incubation for 3 days Cells are aerobic, Gram-staining-positive, non-flagella­ on R2A agar at 30℃. In the API 20NE system, positive ted and rod shaped. Colonies are filamentous, raised, reaction for hydrolysis of esculin and gelatin, β-galacto­ undulate and brown colored after incubation for 3 days sidase activity and utilization of D-glucose, L-arabinose, on R2A agar at 37℃. In the API 20NE system, positive D-mannitol, potassium gluconate and adipic acid. In the reaction for nitrate reduction, arginine dihydrolase, urease API 20NE system, negative reaction for nitrate reduction, activity, hydrolysis of esculin and gelatin, β-galactosidase indole production, glucose fermentation, arginine dihydro- activity (weak) and utilization of D-glucose, L-arabinose lase, urease activity, oxidase activity and utilization of (weak), D-mannitol, N-acetyl-glucosamine, D-maltose D-mannose, N-acetyl-glucosamine, D-maltose, capric acid, (weak), potassium gluconate, malic acid (weak) and phe­ malic acid, trisodium citrate and phenylacetic acid. Strain = nylacetic acid (weak). In the API 20NE system, negative F-111 ( NIBRBAC000503378) was isolated from soil in ′ ″ ′ ″ reaction for indole production, glucose fermentation and Daejeon, Korea (36°22 33.4 N 127°20 33.8 E). utilization of D-mannose, capric acid, adipic acid and tri­ sodium citrate. Strain R-21 (=NIBRBAC000503389) was Description of MMS19-T31 isolated from soil in Gurye, Jeollanam Province, Korea Cells are aerobic, Gram-staining-positive, non-flagel­ ′ ″ ′ ″ (35°11 16.0 N 127°33 02.5 E). lated and rod shaped. Colonies are circular, convex, erose and brown colored after incubation for 3 days on R2A Description of Streptomyces populi DS-12 agar at 30℃. In the API 20NE system, positive reaction Cells are aerobic, Gram-staining-positive, non-flagella­ for hydrolysis of esculin and gelatin, β-galactosidase acti­ ted and filamentous or rod shaped. Colonies are circular, vity (weak) and utilization of D-glucose, L-arabinose raised, erose and light gray colored after incubation for 3 (weak), D-mannose, D-mannitol, N-acetyl-glucosamine, days on R2A agar at 30℃. In the API 20NE system, posi­ D-maltose, potassium gluconate and malic acid. In the tive reaction for nitrate reduction, hydrolysis of esculin API 20NE system, negative reaction for nitrate reduction, and gelatin, β-galactosidase activity and utilization of D- indole production, glucose fermentation, arginine dihydro- glucose, L-arabinose, D-mannose, N-acetyl-glucosamine, lase, urease activity, oxidase activity and utilization of D-maltose, potassium gluconate, adipic acid, malic acid, capric acid, adipic acid, trisodium citrate and phenylacetic trisodium citrate and phenylacetic acid. In the API 20NE acid. Strain MMS19-T31 (=NIBRBAC000503372) was system, negative reaction for indole production, glucose isolated from soil in Gurye, Jeollanam Province, Korea fermentation, arginine dihydrolase, urease activity, oxi­ (35°16′25.9″N 127°28′34.5″E). dase activity and utilization of D-mannitol and capric acid.

Strain DS-12 (=NIBRBAC000503377) was isolated from Description of Streptomyces tanashiensis EAC17 soil in Daejeon, Korea (36°22′35.4″N 127°20′37.2″E). Cells are aerobic, Gram-staining-positive, non-flagella­ Description of 19D1T8 ted and rod shaped. Colonies are circular, smooth, convex, opaque and gray colored after incubation for 3 days on Cells are aerobic, Gram-staining-positive, non-flagella­ R2A agar at 25℃. In the API 20NE system, positive reac­ ted and filamentous. Colonies are circular, convex, umbo­ tion for nitrate reduction, hydrolysis of esculin and gela­ nate and cream colored after incubation for 4 days on MH tin, β-galactosidase activity and utilization of D-glucose, agar at 30℃. In the API 20NE system, positive reaction N-acetyl-glucosamine, D-maltose, potassium gluconate, for hydrolysis of esculin and gelatin, β-galactosidase acti­ malic acid and trisodium citrate. In the API 20NE system, vity, oxidase activity and utilization of D-glucose, L-arabi­ negative reaction for indole production, glucose fermen­ nose, D-mannose, D-mannitol, N-acetyl-glucosa­mine, tation, arginine dihydrolase, urease activity, oxidase acti­ D-maltose, potassium gluconate, adipic acid, malic acid, vity and utilization of L-arabinose, D-mannose, D-manni­ trisodium citrate and phenylacetic acid. In the API 20NE tol, capric acid, adipic acid and phenylacetic acid. Strain system, negative reaction for nitrate reduction, indole EAC17 (=NIBRBAC000503284) was isolated from soil production, glucose fermentation, arginine dihydrolase, in Chungju, Chungcheongbuk Province, Korea (37°08′ urease activity and utilization of capric acid. Strain 19D1T8 23.8″N 127°54′41.8″E). (=NIBRBAC000503275) was isolated from soil around the Dong River, Gangwon Province, Korea (37°22′56.8″N 128°40′01.2″E). Description of MMS19-T12 Cells are aerobic, Gram-staining-positive, non-flagella­ Description of Streptomyces pseudovenezuelae F-111 ted and filamentous or rod shaped. Colonies are circular, Cells are aerobic, Gram-staining-positive, non-flagella­ convex, erose and pink colored after incubation for 3 days 44 JOURNAL OF SPECIES RESEARCH Vol. 10, No. 1 on R2A agar at 30℃. In the API 20NE system, positive References reaction for nitrate reduction, arginine dihydrolase, urease activity, hydrolysis of esculin and gelatin and utilization of Bae, K.S., M.S. Kim, J.H. Lee, J.W. Kang, D.I. Kim, J.H. Lee D-glucose, D-mannose, N-acetyl-glucosamine, D-maltose and C.N. Seong. 2016. Korean indigenous bacterial spe­ and malic acid. In the API 20NE system, negative reaction cies with valid names belonging to the phylum Actinobac- for indole production, glucose fermentation, β-galactosi­ teria. J Microbiol 54(12):789-795. dase activity and utilization of L-arabinose, D-mannitol, Choi, J.H., J.H. Cha, J.W. Bae, J.C. Cho, J. Chun and others. potassium gluconate, capric acid, adipic acid, trisodium 2016. Report on 31 unrecorded bacterial species in Korea citrate and phenylacetic acid. Strain MMS19-T12 (=NIBR that belong to the phylum Actinobacteria. J Sp Res 5(1):1- BAC000503384) was isolated from soil in Gurye, Jeolla­ 13. nam Province, Korea (35°16′25.9″N 127°28′34.5″E). Chun, J. and M. Goodfellow. 1995. A phylogenetic analysis of the genus Nocardia with 16S rRNA gene sequences. Int J Description of Streptomyces zaomyceticus 19D1A31 Syst Bacteriol 45(2):240-245. Dangel, A., A. Berger, R. Konrad and A. Sing. 2019. NGS- Cells are aerobic, Gram-staining-positive, non-flagel­ based phylogeny of -related pathogenicity fac­ lated and filamentous. Colonies are circular, flat and dark tors in different Corynebacterium spp. implies species- cream colored after incubation for 4 days on MH agar at specific virulence transmission. BMC Microbiol 19(1): 30℃. In the API 20NE system, positive reaction for nit­ 28. rate reduction, urease activity, hydrolysis of esculin and Felsenstein, J. 1981. Evolutionary trees from DNA sequences: gelatin (weak), β-galactosidase activity and utilization of a maximum likelihood approach. J Mol Evol 17(6):368- D-glucose, L-arabinose, D-mannose, D-mannitol, N-ace­ 376. tyl-glucosamine, D-maltose, potassium gluconate, adipic Felsenstein, J. 1985. Confidence limit on phylogenies: an appro- acid, malic acid, trisodium citrate and phenylacetic acid. In ­ach using the bootstrap. Evolution 39(4):783-791. the API 20NE system, negative reaction for indole produc­ Fitch, W.M. 1971. Toward defining the course of evolution: tion, glucose fermentation, arginine dihydrolase, oxidase minimum change for a specific tree topology. Syst Zool activity and utilization of capric acid. Strain 19D1A31 20(4):406-416. (=NIBRBAC000503273) was isolated from soil around Goodfellow, M. 2012. Phylum XXVI. Actinobacteria phyl. the Dong River, Gangwon Province, Korea (37°22′56.8″N nov. In: Goodfellow, M., P. Kämpfer, H.-J. Busse, M.E. 128°40′01.2″E). Trujillo, K. Suzuki, W. Ludwig, Whitman, W.B. (eds), Bergey’s Manual of Systematic Bacteriology, second edi­ Description of Collinsella aerofaciens LPB0332 tion, vol. 5, Springer, New York. pp. 33-34. Goodfellow, M. and S.T. Williams. 1983. Ecology of Actino- Cells are anaerobic, Gram-staining-positive, non-flag­ mycetes. Annu Rev Microbiol 37:189-216. ellated and ovoid shaped. Colonies are circular, convex, Hall, T.A. 1999. BioEdit: a user-friendly biological sequence entire and cream colored after incubation for 3 days on alignment editor and analysis program for Windows 95/98/ anaerobe basal medium at 30℃. In the API 20A system, NT. Nucleic Acids Symp Ser 41:95-98. positive reaction for esculin hydrolysis and acid production Hwang, I.S., E.J. Oh, H.B. Lee and C.S. Oh. 2019. Functional from D-mannitol, sucrose, D-maltose, salicin, D-xylose, Characterization of Two Cellulase Genes in the Gram-Posi- L-arabinose, glycerol, D-cellobiose, D-melezitose, D- tive Pathogenic Bacterium Clavibacter michiganensis for raffinose, D-sorbitol, D-rhamnose and D-trehalose. In the Wilting in Tomato. Mol Plant Microbe Interact 32(4):491- API 20A system, negative reaction for oxidase activity, in­ 501. dole formation, urease activity, gelatin hydrolysis and acid Jukes, T.H. and C.R. Cantor. 1969. Evolution of protein mole­ production from D-glucose, D-lactose and D-mannose. cules. In: Munro, H.N. (eds.), Mammalian Protein Meta­ = Strain LPB0332 ( NIBRBAC000503339) was isolated bolism. Academic Press, New York. pp. 21-132. from the intestine of a laboratory mouse in Daejeon, Korea Kim, M.S., J.H. Lee, J.W. Kang, S.B. Kim, J. C. Cho and others. (36°23′56.11″N 127°23′41.76″E). 2016. A report of 38 unrecorded bacterial species in Korea, belonging to the phylum Actinobacteria. 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the class Actinobacteria in Korea. J Sp Res 8(1): 97-108. the actinobacteria. Mol Biol Evol 25(1):1-4. Ko, K.S., C.J. Cha, W.T. Im, S.B. Kim, C.N. Seong and others. Tamura, K., G. Stecher, D. Peterson, A. Filipski and S. Kumar. 2017. A report of 34 unrecorded bacterial species in Korea, 2013. MEGA6: Molecular Evolutionary Genetics Analysis belonging to the Actinobacteria. J Sp Res 6(1):1-14. version 6.0. Mol Biol Evol 30(12):2725-2729. Lee, N.Y., C.J. Cha, W.T. Im, S.B. Kim, C.N. Seong and others. Thompson, J.D., D.G. Higgins and T.J. Gibson. 1994. 2018. A report of 42 unrecorded actinobacterial species in CLUSTAL W: improving the sensitivity of progressive Korea. J Sp Res 7(1):36-49. multiple sequence alignment through sequence weighting, Qin, S., K. Xing, J.H. Jiang, L.H. Xu and W.J. Li. 2011. Bio­ position-specific gap penalties and weight matrix choice. diversity, bioactive natural products and biotechnological Nucleic Acids Res 22(22):4673-4680. potential of plant-associated endophytic actinobacteria. Appl Yoon, S.H., S.M. Ha, S. Kwon, J. Lim, Y. Kim, H. Seo and Microbiol Biotechnol 89(3):457-473. J. Chun. 2017. Introducing EzBioCloud: a taxonomi­ Saitou, N. and M. Nei. 1987. The neighbor-joining method: cally united database of 16S rRNA gene sequences and a new method for reconstructing phylogenetic trees. Mol whole-genome assemblies. Int J Syst Evol Microbiol Biol Evol 4(4):406-425. 67(5):1613-1617. Salam, N., J.Y. Jiao, X.T. Zhang and W.J. Li. 2020. Update on the classification of higher ranks in the phylum Actinobac- teria. Int J Syst Evol Microbiol 70(2):1331-1355. Submitted: November 6, 2020 Servin, J.A., C.W. Herbold, R.G. Skophammer and J.A. Lake. Revised: January 20, 2021 2008. Evidence excluding the root of the tree of life from Accepted: January 22, 2021