A Report of 42 Unrecorded Actinobacterial Species in Korea

Journal36 of Species Research 7(1):36-49, 2018Journal of Species Research Vol. 7, No. 1

A report of 42 unrecorded actinobacterial species in Korea

Na-Young Lee1, Chang-Jun Cha2, Wan-Taek Im3, Seung-Bum Kim4, Chi-Nam Seong5, Jin-Woo Bae6, Kwang Yeop Jahng7, Jang-Cheon Cho8, Kiseong Joh9, Che Ok Jeon10, Hana Yi11 and Soon Dong Lee1,*

1Faculty of Science Education, Jeju National University, Jeju 63243, Republic of Korea 2Department of Biotechnology, Chung-Ang University, Anseong 17546, Republic of Korea 3Department of Biotechnology, Hankyong National University, Anseong 17579, Republic of Korea 4Department of Microbiology and Molecular Biology, Chungnam National University, Daejeon 34134, Republic of Korea 5Department of Biology, Sunchon National University, Suncheon 57922, Republic of Korea 6Department of Biology, Kyung Hee University, Seoul 02447, Republic of Korea 7Department of Life Sciences, Chonbuk National University, Jeonju 28644, Republic of Korea 8Department of Biological Sciences, Inha University, Incheon 22212, Republic of Korea 9Department of Bioscience and Biotechnology, Hankuk University of Foreign Studies, Gyeonggi 17035, Republic of Korea 10School of Biological Science, Chung-Ang University, Seoul 06974, Republic of Korea 11School of Biosystems & Biomedical Science and Department of Public Health Science, Korean University Guro Hospital, Korea University, Seoul 08308, Republic of Korea

*Correspondent: [email protected]

During a study to discover indigenous prokaryotic species in Korea in 2016, a total of 42 actinobacterial isolates were recovered from various environmental samples collected from natural cave, squid, sewage, sea water, trees, droppings of birds, freshwater, eelgrass, mud flat, sediment and soil. On the basis of a tight phylogenetic clade with the closest species and high level of 16S rRNA gene sequence similarity, it was shown that each isolate was assigned to independent and previously described bacterial species which were assigned to the phylum Actinobacteria. The following 42 species have not been reported in Korea: eight species in two genera n the order Corynebacteriales, 26 species of 16 genera in the Micrococcales, one species of one genus in the Micromonosporales, one species of one genus in the Propionibacteriales, four species of two genera in the Streptomycetales and two species of two genera in the Streptosporangiale. Cell morphology, Gram staining reaction, colony colors and features, the media and conditions of incubation, physiological and biochemical characteristics, origins of isolation and strain IDs of 42 unrecorded actinobacterial species are presented in the species description. Keywords: 16S rRNA gene, indigenous prokaryotic species, unrecorded actinobacterial species

Introduction The phylum Actinobacteria (Ludwig et al., 2012) con- sists of classes Actinobacteria, Acidimicrobiia, Corio- Actinobacteria are generally described as aerobic, bacteriia, Nitriliruptoria, Rubrobacteria and Thermo- high-G+C-content, Gram-staining-positive bacteria leophilia based on 16S rRNA gene sequences. Among and contain morphologically diverse groups that ranged these, the class Actinobacteria contains 15 orders: from coccoids or rods to hyphal forms. The branched, Actinomycetales, Actinopolysporales, Bifidobacteria- filamentous hyphae may either remain stable or split into les, Catenulisporales, Corynebacteriales, Frankiales, smaller fragments (Lechevalier & Lechevalier, 1981). In Glycomycetales, Jiangellales, Kineosporiales, Micro- many mycelium-forming actinobacteria, the hyphae are monosporales, Micrococcales, Propionibacteriales, differentiated into aerial mycelia, as well as vegetative Pseudonocardiales, Streptomycetales and Streptospo- mycelia, that bear spores that may by located on the tips rangiales. Members of these bacteria are widely distrib- of filaments or inside sporangia (Lechevalier, 1989; En- uted in nature and occur in both terrestrial and marine sign et al., 1992; Prescott et al., 2002). habitats such as soil, fresh or sea water, plants, insects, February 2018 Lee et al. - Unrecorded actinobacterial species in Korea 37 manure and compost. Owing to their saprophytic nature, 20% glycerol suspensions (or including 2% sea salts) at actinobacteria play an important role as decomposers by -80°C and as lyophilized ampoules. degrading a wide range of organic compounds and thus Gram staining was performed using a Gram-stain- contribute to nutrient cycling in natural environments ing kit (bioMérieux) following the manufacturer’s in- through the turnover of complex organic materials structions. The morphology and color of colonies were (Lechevalier & Lechevalier, 1981; Williams et al., 1984; observed on agar plates showing optimal growth. Cell Mincer et al., 2002; Prescott et al., 2002). Actinobacte- morphology was observed using either transmission or ria are of significant industrial importance as producers scanning electron microscopy. Utilization of sole carbon of a wide range of secondary metabolites such as anti- and energy sources and biochemical properties and were biotics, anti-tumor agents, immunomodulatory agents tested with API 20NE kit (bioMérieux) according to the and other industrial useful enzymes (Nolan and Cross, manufacturer’s instructions. 1988; Mincer et al., 2002; Prescott et al., 2002; Busti et Bacterial DNA extraction, PCR amplification of the al., 2006). Many actinobacteria form symbiotic relation- 16S rRNA gene by PCR and its sequencing were per- ships with plants and insects, and reside as endophytes formed using the standard procedures described else- by providing beneficial secondary metabolites to the where. The 42 16S rRNA gene sequences of the actino- host plants through the production of antimicrobial mol- bacterial isolates were aligned with the corresponding ecules or plant growth promoters (Coombs & Franco, sequences of closely related bacterial species retrieved 2003; Hasegawa et al., 2006; Zhang et al., 2007). Some from the EzTaxon-e server (Kim et al., 2012), using the actinobacteria may also be harmful and cause disease in Clustal_X program (Thompson et al., 1997). After re- human, animals and plants (Lechevalier & Lechevalier, moval of gaps and manual optimization according to the 1981; Williams et al., 1989; Prescott et al., 2002) secondary structure of Escheichia coli, the sequences During an investigation of bacterial species indige- were used for phylogenetic analysis. A neighbor-joining nous to Korea that was performed through a research (Saitou and Nei, 1987) tree was drawn using evolution- program supported by the National Institute of Biolog- ary distances generated using the model of Jukes & Can- ical Resources (NIBR) in 2016, a large number of un- tor (1969). Phylogenetic analyses were also performed recorded species in Korea were recovered from diverse using maximum-parsimony (Fitch, 1971) and maxi- environmental samples. In this study, we identified and mum-likelihood (Felsenstein, 1981) methods. Bootstrap classified bacterial isolates that belonged to the phylum analysis (Felsenstein, 1985) was performed for estimat- Actinobacteria and describe 42 unrecorded actinobac- ing reliability of tree topology, based on 1,000 replica- terial species that have not been previously reported in tions. Korea.

Results and Discussion Materials and Methods The 42 isolates were recovered from various environ- A total of 42 bacterial strains in the phylum Acti- ments and were found to belong to the phylum Actino- nobacteria were isolated from various environmental bacteria. They were distributed in six orders of the class samples collected from natural cave, lava cave, squid, Actinobacteria: eight strains in the order Corynebacte- sewage, sea water, tree, droppings of birds, freshwater, riales, 26 strains in the Micrococcales; one strain in the eelgrass, mud flat, sediment and soils (Table 1). For bac- Micromonosporales; one strain in the Propionibacteria- terial isolation, the environmental samples were treated les; four strains in the Streptomycetales; and two strains independently in several laboratories. Pure cultures were in the Streptosporangiales (Table 1). All the strains obtained after subcultures were attempted several times were Gram-staining-positive, non-motile and most were and grown at 20-37°C for 1-22 days on varied culture represented by coccoid, oval or rod-shaped morphol- media including R2A agar (BD), marine agar 2216 (MA; ogy (Fig. 1). The neighbor-joining tree (Fig. 2) based BD), tryptic soy agar (TSA; BD), nutrient agar (NA; on 16S rRNA gene sequences shows the phylogenetic BD), ISP (International Streptomyces Project) 2 agar relationships of the strains of the order Corynebacteri- (Shirling & Gottlieb, 1966), Mueller Hinton medium ales: Mycobacterium (2 species) of the Mycobacteriace- (BD) and starch-casein agar (SCA; Küster and Williams, ae and Gordonia (2 species); and Nocardia (3 species) 1964), depending on the strains. The strain numbers, and Rhodococcus (1 species) of the Nocardiaceae. The designated NIBR numbers, most closely related species, strains of the order Micrococcales were assigned to 16 origins of isolation, culture media and incubation condi- genera in eight families: Demequina (1 species) of the tions of 42 unrecorded actinobacterial species are sum- family Demequinaceae; Brachybacterium (2 species) marized in Table 1. The pure cultures were stored as 10- and Demacoccus (1 species) of the Dermabacteraceae; 38 Journal of Species Research Vol. 7, No. 1 con. I ncubation 30° C , 2 days 30° C , 2 days 30° C , 3 days 30° C , 4 days 25° C , 3 days 30° C , 5 days 25° C , 3 days 37° C , 2 days 25° C , 1 day 30° C , 2 days 30° C , 2 days 25° C , 3 days 30° C , 2 days 25° C , 3 days 20° C , 2 days 24° C , 3 days 25° C , 1 day 20° C , 2 days 25° C , 1 day 30° C , 3 days 30° C , 2 days 25° C , 1 day 30° C , 5 days 30° C , 5 days 37° C , 2 days 30° C , 2 days 25° C , 3 days 30° C , 5 days 30° C , 22 days 30° C , 22 days 25° C , 1 day 30° C , 3 days 25° C , 3 days 30° C , 3 days 30° C , 3 days 25° C , 3 days 30° C , 2 days 37° C , 2 days 37° C , 2 days 25° C , 3 days 25° C , 3 days 20° C , 2 days (p H 5) (p H 5) (p H 5) (2x)

Medium TS A R 2 A R 2 A S CA NA R 2 A I S P 2 S CA R 2 A M A M A R 2 A R 2 A TS A R 2 A R 2 A R 2 A M A M A M A M A M A M A R 2 A M A I S P 2 NA Mueller H inton R 2 A R 2 A I S P 2 I S P 2 M A R 2 A TS A I S P 2 TS A TS A M A R 2 A R 2 A R 2 A I solation source Soil Sediment Tree Soil Yongcheon C ave Yongcheon Ginseng soil C ave Yongcheon Soil Ginseng soil Droppings of spoonbills Squid Sediment Sediment Mud flat Ginseng soil Sewage Ginseng soil Droppings of eagles E elgrass Squid Droppings of eagles Squid Mud flat Sediment Yongcheon C ave Yongcheon Droppings of eagles Squid C ave Yongcheon Sewage Sea water N atural cave N atural cave Squid Soil Ginseng soil Soil Droppings of eagles Droppings of spoonbills Ginseng soil F reshwater F reshwater Droppings of eagles

99 99 100 100 100 100 100 99.5 99.7 99.7 99.9 99.6 98.8 98.9 (%) 99.9 99.8 99.9 99.6 98.8 99.6 99.1 99.7 99.4 99.3 99.3 99.7 99.1 99.6 99.8 99.5 98.8 98.9 99.7 99.4 99.3 99.9 99.8 99.3 99.2 98.8 99.9 98.7 Similarity

( N ame of type strain) Most closely related species Agromyces italicus Agromyces Gryllotalpicola reticulitermitis Jonesia quinghaiensis Leifsonia poae Oerskovia turbata echinospora Micromonospora fastidiosum Aeromicrobium Kitasatospora kifunensis Intrasporangium oryzae Cellulosimicrobium cellulans Cellulosimicrobium Brachybacterium muris Brachybacterium rhamnosum Dermacoccus nishinomiyaensis Gordonia aichiensis Gordonia Nonomuraea harbinensis Arthrobacter niigatensis Arthrobacter Arthrobacter nicotinovorans Arthrobacter nicotianae Arthrobacter gandavensis Arthrobacter rhombi Arthrobacter Kocuria carniphila lylae Micrococcus lutea Nesterenkonia Renibacterium salmoninarum Nocardia soli Nocardia fluminea Nocardia vermiculata Nocardia Rhodococcus ruber Demequina flava Gordonia terrae Gordonia Mycobacterium algericum Mycobacterium aubagnense Nocardiopsis synnemataformans Nocardiopsis Streptomyces kurssanovii Streptomyces Streptomyces durhamensis Streptomyces Streptomyces blastmyceticus Streptomyces Microbacterium esteraromaticum Microbacterium Microbacterium testaceum Microbacterium Microbacterium aerolatum Microbacterium fluvii Microbacterium hatanonis Microbacterium Microbacterium resistens Microbacterium NI B R I D NI B R AC 000498411 NI B R AC 000498614 NI B R AC 000498454 NI B R AC 000498569 NI B R AC 000498620 NI B R AC 000498651 NI B R AC 000498604 NI B R AC 000498650 NI B R AC 000498624 NI B R AC 000498587 NI B R AC 000498393 NI B R AC 000498523 NI B R AC 000498570 NI B R AC 000498577 NI B R AC 000498659 NI B R AC 000498588 NI B R AC 000498596 NI B R AC 000498399 NI B R AC 000498480 NI B R AC 000498521 NI B R AC 000498398 NI B R AC 000498515 NI B R AC 000498539 NI B R AC 000498568 NI B R AC 000498652 NI B R AC 000498653 NI B R AC 000498386 NI B R AC 000498431 NI B R AC 000498631 NI B R AC 000498517 NI B R AC 000498662 NI B R AC 000498661 NI B R AC 000498516 NI B R AC 000498617 NI B R AC 000498585 NI B R AC 000498623 NI B R AC 000498378 NI B R AC 000498381 NI B R AC 000498599 NI B R AC 000498634 NI B R AC 000498635 NI B R AC 000498394 Strain I D TS3307 UL 003 H M F 7422 5189 MMS16- CNU 056 Y C 4-25 Gsoil 1174 Y C 3-14 MMS16- CNU 292 Gsoil 608 P M A 3209 LP B0135 6228 10038 B E 5-3 Gsoil 1046 J52 Gsoil 1513 P M A 2201 Z O D2-24 LP B0133 P M A 2305 LP B0125 I M CC 25640 5187 Y C 6-12 Y C 7-20 AR 23308 Y H J2 JGM01 LP B0127 C 6-3 C 1-45 LP B0126 UL 451 A Gsoil 224 MMS16- CNU 183 P TS2402 Gsoil 163 CAH -3 2JSS04 A M 3210 Genus Agromyces Agromyces Gryllotalpicola Jonesia Leifsonia Oerskovia Micromonospora Aeromicrobium Kitasatospora Intrasporangium Cellulosimicrobium Cellulosimicrobium Brachybacterium Brachybacterium Dermacoccus Gordonia Gordonia Nonomuraea Arthrobacter Arthrobacter Arthrobacter Arthrobacter Arthrobacter Arthrobacter Arthrobacter Kocuria Micrococcus Nesterenkonia Renibacterium Nocardia Nocardia Nocardia Nocardia Rhodococcus Demequina Gordonia Gordonia Mycobacterium Mycobacterium Nocardiopsis Nocardiopsis Streptomyces Streptomyces Microbacterium Streptomyces Streptomyces Streptomyces Microbacterium Microbacterium Microbacterium Microbacterium Microbacterium Microbacterium F amily Microbacteriaceae Jonesiaceae Sanguibacteraceae Micromonosporaceae Nocardioidaceae Streptomycetaceae Intrasporangiaceae Promicromonosporaceae Dermabacteraceae Nocardiaceae Streptosporangiaceae Micrococcaceae Demequinaceae Mycobacteriaceae Nocardiopsaceae Summary of the isolated strains belonging to the Actinobacteria and their taxonomic affiliations. Summary of the isolated strains belonging to O rder Actinobacteria Micromonosporales Propionibacteriales Streptomycetales Micrococcales Table 1. Table C lass Corynebacteriales Streptosporangiales February 2018 Lee et al. - Unrecorded actinobacterial species in Korea 39

(1) (2) (3) (4) (5) (6)

(7) (8) (9) (10) (11) (12)

(13) (14) (15) (16) (17) (18)

(19) (20) (21) (22) (23) (24)

(25) (26) (27) (28) (29) (30)

(31) (32) (33) (34) (35)

Fig. 1. Transmission electron micrographs or scanning electron micrographs of cells of the strains isolated in this study. Strains: 1, C1-45; 2, C6-3; 3, BE5-3; 4, LPB0127; 5, YC6-12; 6, YC7-20; 7, AR23308; 8, YHJ2; 9, JGM01; 10, LPB0135; 11, 6228; 12, 10038; 13, Gsoil 608; 14, 5189; 15, UL003; 16, HMF7422; 17, MMS16-CNU056; 18, ATS3307; 19, PTS2402; 20, AMA3210; 21, Gsoil 163; 22, CAH-3; 23, 2JSS04; 24, J52; 25, Gsoil 1513; 26, PMA2201; 27, ZOD2-24; 28, LPB0133; 29, PMA2305; 30, LPB0125; 31, IMCC25640; 32, 5187; 33, PMA3209; 34, YC4-25; 35, YC3-14.

Intrasporangium (1 species) of the Intrasporangiaceae; losimicrobium (1 species) of the Promicromonosporace- Jonesia (1 species) of the Jonesiaceae; Agromyces (1 ae; and Oerskovia (1 species) of the Sanguibacteraceae species), Gryllotalpicola (1 species), Leifsonia (1 spe- and their phylogenetic relationships were shown in Fig. cies) and Microbacterium (6 species) of the Microbac- 3. The order Streptomycetales consisted of four isolates teriaceae; Arthrobacter (5 species), Kocuria (1 species), that were distributed in the genera Kitasatospora (1 spe- Micrococcus (1 species), Nesterenkonia (1 species) and cies) and Streptomyces (3 species) of the family Strep- Renibacterium (1 species) of the Micrococcaceae; Cellu- tomycataceae. The orders Micromonosporales and Pro- 40 Journal of Species Research Vol. 7, No. 1

Fig. 2. Neighbor-joining phylogenetic tree, based on 16S rRNA gene sequences, showing the relationships between the strains isolated in this study and their relatives of the orders Corynebacteriales in the class Actinobacteria. Asterisks indicate that the corresponding branches were also recovered in both the maximum-likelihood and maximum-parsimony trees. Bootstrap values (>60%) are shown at the branching points. Bar, 0.01 substitutions per nucleotide position. pionibacteriales contained only one isolate that was af- teriales, whose members form coccoids or rods. Some filiated to the genus Micromonospora of the family Mi- mycelium-forming species showed further differentiated cromonosporaceae and to the genus Nocardioidaceae, features such as the formation of spores on the ends of respectively. The order Streptosporangiales comprised aerial mycelia (Lechevalier, 1989; Prescott et al., 2002). two isolates, each of which was assigned to the genus Nocardiopsis of the family Nocardiopsaceae and the ge- Description of Mycobacterium aubagnense C1-45 nus Nonomuraea of the family Stereptosporangiaceae, respectively. The tree given in Fig. 4 reveals the phylo- Cells are Gram-staining-positive, flagellated, non-pig- genetic relationships between the isolates of the above mented and rod-shaped. Colonies are circular, convex, four orders and their closest relatives. Most strains of entire and light brown-colored after 22 days on ISP2 these orders formed well-developed, branched aerial at 30°C. Positive for nitrate reduction in API 20NE but and substrate hypha. Among members of these orders, negative for indole production, glucose fermentation, most produce vegetative and aerial mycelia depending arginine dihydrolase, urease, esculin hydrolysis, gelati- on the species, with exception of the order Propionibac- nase and β-galactosidase. Does not utilize D-glucose, February 2018 Lee et al. - Unrecorded actinobacterial species in Korea 41

Fig. 3. Neighbor-joining phylogenetic tree, based on 16S rRNA gene sequences, showing the relationships between the strains isolated in this study and their relatives of the order Micrococcales in the class Actinobacteria. Asterisks indicate that the corresponding branches were also recovered in both the maximum-likelihood and maximum-parsimony trees. Bootstrap values (>60%) are shown at the branching points. Bar, 0.01 substitutions per nucleotide position.

L-arabinose, D-mannitol, D-mannose, N-acetyl-glu- Description of Mycobacterium algericum C6-3 cosamine, D-maltose, gluconate, caprate, adipate, malate, citrate and phenylacetate. Strain C1-45 ( =NI- Cells are Gram-staining-positive, non-flagellated, BRBA000498661) has been isolated from natural cave, non-pigmented and rod-shaped. Colonies are circular, Jeju, Korea. convex, entire and light brown-colored after 22 days on ISP2 at 30°C. Positive for nitrate reduction, esculin 42 Journal of Species Research Vol. 7, No. 1

Fig. 4. Neighbor-joining phylogenetic tree, based on 16S rRNA gene sequences, showing the relationships between the strains isolated in this study and their relatives of the orders Micromonosporales, Propionibacteriales, Streptomycetales and Streptosporangiales in the class Actinobacteria. Asterisks indicate that the corresponding branches were also recovered in both the maximum-likelihood and maximum-parsimony trees. Bootstrap values (>60%) are shown at the branching points. Bar, 0.01 substitutions per nucleotide position. hydrolysis and β-galactosidase in API 20NE but nega- TSA at 30°C. Positive for nitrate reduction and urease tive for indole production, glucose fermentation, argi- in API 20NE, weakly positive for esculin hydrolysis but nine dihydrolase, urease and gelatinase. Does not uti- negative for indole production, glucose fermentation, lize D-glucose, L-arabinose, D-mannitol, D-mannose, arginine dihydrolase, gelatinase and β-galactosidase. N-acetyl-glucosamine, D-maltose, gluconate, caprate, Does not utilize D-glucose, L-arabinose, D-mannitol, adipate, malate, citrate and phenylacetate. Strain C6-3 D-mannose, N-acetyl-glucosamine, D-maltose, gluco- (=NIBRBA000498662) has been isolated from natural nate, caprate, adipate, malate, citrate and phenylacetate. cave, Jeju, Korea. Strain BE5-3 (=NIBRBA000498659) has been isolated from mud flat, Eulsukdo, Busan, Korea. Description of Gordonia aichiensis BE5-3 Description of Gordonia terrae LPB0127 Cells are Gram-staining-positive, non-flagellated, non-pigmented and rod-shaped. Colonies are circular, Cells are Gram-staining-positive, non-flagellated, convex, entire and orange yellow-colored after 5 days on non-pigmented and rod-shaped. Colonies are circular, February 2018 Lee et al. - Unrecorded actinobacterial species in Korea 43 convex, entire and pink-colored after 1 day on MA at pings of eagles, Seoul Grand Park, Gwacheon, Gyeong- 25°C. Positive for urease in API 20NE but negative for gi, Korea. nitrate reduction, indole production, glucose fermentation, arginine dihydrolase, esculin hydrolysis, gelatinase Description of Rhodococcus ruber YHJ2 and β-galactosidase. D-Mannos and malate are utilized. Cells are Gram-staining-positive, non-flagellated and Does not utilize D-glucose, L-arabinose, D-mannitol, rod/oval-shaped. Colonies are Irregular, raised, entire N-acetyl-glucosamine, D-maltose, gluconate, caprate, and orange-colored after 2 days on R2A at 30°C. Pos- adipate, citrate and phenylacetate. Strain LPB0127 itive for nitrate reduction in API 20NE but negative (=NIBRBAC000498517) has been isolated from squid, for indole production, glucose fermentation, arginine Jumunjin, Gangwon-do, Korea. dihydrolase, urease, esculin hydrolysis, gelatinase and β-galactosidase. D-Glucose, D-mannose and malate Description of Nocardia soli YC6-12 are utilized. Does not utilize L-arabinose, D-mannitol, Cells are Gram-staining-positive, non-flagellated, N-acetyl-glucosamine, D-maltose, gluconate, caprate, non-pigmented and fragmented mycelium. Colonies are adipate, citrate and phenylacetate. Strain YHJ2 ( =NI- circular, convex, entire and whitish pink-colored after 5 BRBAC000498431) has been isolated from sewage, Bu- days on IPS2 at 30°C. Esculin hydrolysis is weakly pos- san, Korea. itive in API 20NE but negative for nitrate reduction, indole production, glucose fermentation, arginine dihydro- Description of Demequina flava JGM01 lase, urease and gelatinase and β-galactosidase. N-Ace- Cells are Gram-staining-positive, non-flagellated and tyl-glucosamine and gluconate are weakly utilized. Does oval-shaped. Colonies are circular, convex and yel- not utilize D-glucose, L-arabinose, D-mannitol, D-man- low-colored after 3 days on R2A at 25°C. Strain JGM01 nose, D-maltose, caprate, adipate, malate, citrate and (=NIBRBA000498631) has been isolated from sea wa- = phenylacetate. Strain YC6-12 ( NIBRBA000498652) ter, Mokji island, Jeju, Korea. has been isolated from Yongcheon Cave, Jeju, Korea. Description of Brachybacterium muris LPB0135 Description of Nocardia fluminea YC7-20 Cells are Gram-staining-positive, non-flagellated, Cells are Gram-staining-positive, non-flagellated, non-pigmented and coccus-shaped. Colonies are cir- non-pigmented and fragmented mycelium. Colonies cular, convex, entire and yellow-colored after 1 day on are circular, convex, entire and whitish pink-colored MA at 25°C. Positive for urease and β-galactosidase after 5 days on NA at 30°C. Esculin hydrolysis is in API 20NE but negative for nitrate reduction, indole weakly positive in API 20NE but negative for nitrate production, glucose fermentation, arginine dihydro- reduction, indole production, glucose fermentation, lase, esculin hydrolysis, gelatinase. Does not utilize arginine dihydrolase, urease, gelatinase and β-galac- D-glucose, L-arabinose, D-mannitol, D-mannos, N-ace- tosidase. N-Acetyl-glucosamine is weakly utilized. tyl-glucosamine, D-maltose, gluconate, caprate, adi- Does not utilize D-glucose, L-arabinose, D-mannitol, pate, malate, citrate and phenylacetate. Strain LPB0135 D-mannose, D-maltose, gluconate, caprate, adipate, (=NIBRBAC000498523) has been isolated from squid, malate, citrate and phenylacetate. Strain YC7-20 (=NI- Jumunjin, Gangwon-do, Korea. BRBA000498653) has been isolated from Yongcheon Cave, Jeju, Korea. Description of Brachybacterium rhamnosum 6228 Cells are Gram-staining-positive, non-flagellated and Description of Nocardia vermiculata AR23308 coccus-shaped. Colonies are circular, entire, smooth, Cells are Gram-staining-positive, non-flagellated raised and pale-yellow-colored after 2 days on R2A at and rod-shaped. Colonies are circular, raised, hard and 30°C. Positive for glucose fermentation, esculin hydro- cream-colored after 2 days on Mueller Hinton at 37°C. lysis, gelatinase and β-galactosidase in API 20NE but Positive for nitrate reduction, esculin hydrolysis and negative for nitrate reduction, indole production, argi- β-galactosidase in API 20NE but negative for indole nine dihydrolase and urease. D-Glucose, D-mannitol, production, glucose fermentation, arginine dihydrolase, D-mannose, D-maltose, gluconate are utilized. Does urease and gelatinase. D-Maltose, gluconate, adipate, not utilize L-arabinose, N-acetyl-glucosamine, caprate, malate is utilized. Does not utilize D-glucose, L-arabi- adipate, malate, citrate and phenylacetate. Strain 6228 nose, D-mannitol, D-mannose, N-acetyl-glucosamine, (=NIBRBAC000498570) has been isolated from sedi- caprate, citrate and phenylacetate. Strain AR23308 ment, Han River, Korea. (=NIBRBAC000498386) has been isolated from drop- 44 Journal of Species Research Vol. 7, No. 1

Description of Dermacoccus nishinomiyaensis 10038 pate, malate, citrate and phenylacetate. Strain UL003 ( =NIBRBAC000498614) has been isolated from soil, Cells are Gram-staining-positive, flagellated and coc- Ulleung-gun, Gyeongsangbuk-do, Korea. cus-shaped. Colonies are circular, entire, smooth, raised and yellow-colored after 2 days on R2A at 30°C. Pos- Description of Gryllotalpicola reticulitermitis HMF7422 itive for gelatinase and β-galactosidase in API 20NE but negative for nitrate reduction, indole production, Cells are Gram-staining-positive, non-flagellated and glucose fermentation, arginine dihydrolase, urease and irregular oval-shaped. Colonies are Circular, convex, esculin hydrolysis. D-Glucose, L-arabinose, D-man- entire and white-colored after 3 days on R2A at 30°C. nitol, D-mannose, N-acetyl-glucosamine, D-maltose, Positive for esculin hydrolysis and β-galactosidase in gluconate, adipate, malate, citrate and phenylacetate are API 20NE but negative for nitrate reduction, indole utilized. Does not utilize caprate. Strain 10038 ( =NI- production, glucose fermentation, arginine dihydrolase, BRBAC000498577) has been isolated from sediment, urease, gelatinase. D-Glucose, D-mannitol and gluco- Han River, Korea. nate are utilized. Does not utilize L-arabinose, D-mannose, N-acetyl-glucosamine, D-maltose, caprate, adi- Description of Intrasporangium oryzae Gsoil 608 pate, malate, citrate and phenylacetate. Strain HMF7422 ( =NIBRBAC000498454) has been isolated from tree, Cells are Gram-staining-positive, non-flagellated and Yongin, Gyeonggi, Korea. coccus-shaped. Colonies are round, convex, circular and yellow-colored after 3 days on R2A at 25°C. Positive Description of Leifsonia poae MMS16-CNU056 for nitrate reduction, esculin hydrolysis, gelatinase and β-galactosidase in API 20NE but negative for indole Cells are Gram-staining-positive, non-flagellated and production, glucose fermentation, arginine dihydrolase rod-shaped. Colonies are circular, glistering, moist and and urease. D-Glucose, D-mannitol, D-maltose, gluco- white-colored after 3 days on SCA (pH 5) at 30°C. Pos- nate, malate are utilized. Does not utilize L-arabinose, itive for esculin hydrolysis and β-galactosidase in API D-mannose, N-acetyl-glucosamine, caprate, adipate, 20NE but negative for nitrate reduction, indole produc- citrate and phenylacetate. Strain Gsoil 608 ( =NI- tion, glucose fermentation, arginine dihydrolase, urease BRBAC000498587) has been isolated from ginseng and gelatinase. D-Glucose, L-arabinose, D-mannitol, soil, Anseong, Korea. N-acetyl-glucosamine and D-maltose are utilized. Does not utilize D-mannose, gluconate, caprate, adipate, ma- Description of Jonesia quinghaiensis 5189 late, citrate and phenylacetate. Strain MMS16-CNU056 ( =NIBRBAC000498620) has been isolated from soil, Cells are Gram-staining-positive, non-flagellated Daejeon, Korea. and rod-shaped. Colonies are circular, entire, smooth, raised and pale-yellow-colored after 2 days on R2A at Description of Microbacterium esteraromaticum 30°C. Positive for nitrate reduction, esculin hydrolysis, ATS3307 β-galactosidase in API 20NE but negative for indole production, glucose fermentation, arginine dihydrolase, Cells are Gram-staining-positive, non-flagellated urease and gelatinase. D-Glucose, L-arabinose, D-man- and rod-shaped. Colonies are Circular, slightly convex, nitol, D-mannose, D-maltose, gluconate are utilized. smooth, glistening and pale yellow-colored after 2 days Does not utilize N-acetyl-glucosamine, caprate, adipate, of incubation on TSA at 37°C. Positive for esculin hy- malate, citrate and phenylacetate. Strain 5189 ( =NI- drolysis in API 20NE, but negative for nitrate reduction, BRBAC000498569) has been isolated from sediment, indole production, glucose fermentation, arginine dihy- Han River, Korea. drolase, urease, gelatinase and β-galactosidase. D-Glu- cose, L-arabinose, D-mannitol, D-maltose are utilized. Description of Agromyces italicus UL003 Does not utilize, D-mannose, N-acetyl-glucosamine, gluconate, caprate, adipate, malate, citrate and pheny- Cells are Gram-staining-positive, non-flagellated, lacetate. Strain ATS3307 (=NIBRBAC000498378) has non-pigmented and rod-shaped. Colonies are circular, been isolated from droppings of eagles, Seoul Grand convex, entire and cream-colored after 2 days on TSA at Park, Gwacheon, Gyeonggi, Korea. 30°C. Positive for nitrate reduction and β-galactosidase in API 20NE but negative for indole production, glucose Description of Microbacterium testaceum PTS2402 fermentation, arginine dihydrolase, urease, esculin hydrolysis and gelatinase. D-Glucose, D-mannitol, D-man- Cells are Gram-staining-positive, non-flagellated nose, N-acetyl-glucosamine and D-maltose are utilized. and rod-shaped. Colonies are circular, glistening and Does not utilize L-arabinose, gluconate, caprate, adi- cream-colored after 2 days on TSA at 20°C. Positive February 2018 Lee et al. - Unrecorded actinobacterial species in Korea 45 for esculin hydrolysis and β-galactosidase in API 20NE Description of Microbacterium hatanonis 2JSS04 but negative for nitrate reduction, indole production, Cells are Gram-staining-positive, non-flagellated glucose fermentation, urease, arginine dihydrolase and rod-shaped. Colonies are circular, slightly convex, and gelatinase. D-Glucose, L-arabinose, D-mannitol, smooth and yellow-colored after 3-4 days on 2x R2A at D-mannose and D-maltose are utilized. Does not utilize 25°C. Positive for nitrate reduction, esculin hydrolysis N-acetyl-glucosamine, gluconate, caprate, adipate, ma- and β-galactosidase in API 20NE but negative for indole late, citrate and phenylacetate. Strain PTS2402 ( =NI- production, glucose fermentation, arginine dihydrolase, BRBA000498381) has been isolated from droppings of urease and gelatinase. D-Mannose is utilized. Does not spoonbills, Seoul Grand Park, Gwacheon, Gyeonggi, utilize D-glucose, L-arabinose, D-mannitol, N-ace- Korea. tyl-glucosamine, D-maltose, gluconate, caprate, adipate, malate, citrate and phenylacetate. Strain 2JSS04 (=NI- Description of Microbacterium resistens AMA3210 BRBA000498635) has been isolated from freshwater, Cells are Gram-staining-positive, non-flagellated and Jeonju, Korea. rod-shaped. Colonies are circular and yellow-colored after 2 days on MA at 37°C. Positive for nitrate reduction, Description of Arthrobacter niigatensis J52 esculin hydrolysis and β-galactosidase in API 20NE but Cells are Gram-staining-positive, non-flagellated, negative for indole production, glucose fermentation, non-pigmented and rod-shaped. Colonies are circular, arginine dihydrolase, urease and gelatinase. Does not raised, entire and white-colored after 2 days on R2A at utilize D-glucose, L-Arabinose, D-mannitol, D-man- 30°C. Positive for esculin hydrolysis and β-galactosi- nose, N-acetyl-glucosamine, D-maltose, gluconate, dase in API 20NE but negative nitrate reduction, indole caprate, adipate, malate, citrate and phenylacetate. Strain production, glucose fermentation, arginine dihydrolase, = AMA3210 ( NIBRBAC000498394) has been isolated urease and gelatinase. D-Glucose, D-mannitol, D-man- from droppings of eagles, Seoul Grand Park, Gwacheon, nose, D-maltose, gluconatem and malate are utilized. Gyeonggi, Korea. Does not utilize L-arabinose, N-acetyl-glucosamine, caprate, adipate, citrate and phenylacetate. Strain J52 Description of Microbacterium aerolatum Gsoil 163 ( =NIBRBAC000498411) has been isolated from sew- Cells are Gram-staining-positive, non-flagellated age, Busan, Korea. and rod-shaped. Colonies are convex, smooth and ivo- ry-colored after 3 days on R2A at 25°C. Positive for Description of Arthrobacter nicotinovorans Gsoil 1513 esculin hydrolysis and β-galactosidase in API 20NE Cells are Gram-staining-positive, non-flagellated and but negative for nitrate reduction, indole production, rod-shaped. Colonies are convex, circular, smooth and glucose fermentation, arginine dihydrolase, urease pale-yellow-colored after 3 days on R2A at 25°C. Pos- and gelatinase. D-Glucose, L-arabinose, D-mannitol, itive for esculin hydrolysis, gelatinase and β-galacto- D-mannose, N-acetyl-glucosamine, D-maltose, gluco- sidase in API 20NE but negative for nitrate reduction, nate, malate, citrate and phenylacetate are utilized. Does indole production, glucose fermentation, arginine di- not utilize caprate, adipate. Strain Gsoil 163 ( =NI- hydrolase and urease. D-Glucose, L-arabinose, D-man- BRBAC000498599) has been isolated from ginseng nitol, D-mannose, N-acetyl-glucosamine, D-maltose, soil, Anseong, Korea. gluconate, malate, citrate and phenylacetate are utilized. Does not utilize caprate, adipate. Strain Gsoil 1513 Description of Microbacterium fluvii CAH-3 ( =NIBRBAC000498596) has been isolated from ginseng soil, Anseong, Korea. Cells are Gram-staining-positive, non-flagellated and rod and oval-shaped. Colonies are circular, convex, en- Description of Arthrobacter nicotianae PMA2201 tire and yellow-colored after 3 days on R2A at 25°C. Positive for esculin hydrolysis and β-galactosidase in Cells are Gram-staining-positive, non-flagellated and API 20NE but negative for nitrate reduction, indole rod-shaped. Colonies are circular and cream-colored production, glucose fermentation, arginine dihydro- after 2 days on MA at 20°C. Positive for nitrate reduc- lase, urease and gelatinase. D-Glucose, D-mannose and tion, urease, gelatinase and β-galactosidase but negative D-maltose and are utilized. Does not utilize L-arabinose, for indole production, glucose fermentation, arginine D-mannitol, N-acetyl-glucosamine, gluconate, caprate, dihydrolase and esculin hydrolysis. D-Glucose, L-arab- adipate, malate, citrate and phenylacetate. Strain CAH-3 inose, D-mannose, N-acetyl-glucosamine, D-maltose, ( =NIBRBA000498634) has been isolated from fresh- D-mannitol, gluconate, caprate, adipate, malate, citrate water, Jeonju, Korea. and phenylacetate are utilized. Strain PMA2201 (=NI- 46 Journal of Species Research Vol. 7, No. 1

BRBAC000498399) has been isolated from droppings drolysis and β-galactosidase. gluconate, adipate, malate, of eagles, Seoul Grand Park, Gwacheon, Gyeonggi, Ko- citrate are utilized. D-Glucose, L-arabinose, D-mannitol, rea. D-mannose, N-acetyl-glucosamine, D-maltose, caprate and phenylacetate are not utilized. Strain LPB0125 Description of Arthrobacter gandavensis ZOD2-24 (=NIBRBAC000498515) has been isolated from squid, Jumunjin, Gangwon-do, Korea. Cells are Gram-staining-positive, non-flagellated and rod-shaped. Colonies are opaque, circular, smooth, low Description of Nesterenkonia lutea IMCC25640 convex and pale-yellow-colored after 3 days on MA at 24°C. Positive for nitrate reduction and β-galactosidase Cells are Gram-staining-positive, non- flagellated and in API 20NE but negative for indole production, glu- rod-shaped. Colonies are circular, convex, entire and cose fermentation, arginine dihydrolase, urease, esculin yellow-colored after 3 days on MA at 20°C. Negative hydrolysis and gelatinase. D-Glucose, L-arabinose, for nitrate reduction, indole production, glucose fermen- D-mannitol, D-mannose, N-acetyl-glucosamine, D-malt- tation, arginine dihydrolase, urease, esculin hydrolysis, ose, gluconate, caprate, adipate, malate, citrate and gelatinase and β-galactosidase in API 20NE. D-Glucose phenylacetate are not utilized. Strain ZOD2-24 ( =NI- is utilized. Does not utilize L-arabinose, D-mannitol, BRBAC000498480) has been isolated from eelgrass, D-mannos, N-acetyl-glucosamine, D-maltose, gluco- Seosan, Chungcheongnam-do, Korea. nate, caprate, adipate, malate, citrate and phenylacetate. Strain IMCC25640 (=NIBRBAC000498539) has been Description of Arthrobacter rhombi LPB0133 isolated from mud flat, Yeongjongdo island, Incheon, Korea. Cells are Gram-staining-positive, flagellated, non-pigmented and rod-shaped. Colonies are circular, convex, Description of Renibacterium salmoninarum 5187 entire and yellow-colored after 1 day on MA at 25°C. Negative for nitrate reduction, indole production, glu- Cells are Gram-staining-positive, non-flagellated and cose fermentation, arginine dihydrolase, urease, esculin rod-shaped. Colonies are circular, entire, dry, flat and hydrolysis, gelatinase and β-galactosidase in API 20NE. white-colored after 2 days on R2A at 30°C. Positive for Does not utilize D-glucose, L-arabinose, D-mannitol, esculin hydrolysis, gelatinase and β-galactosidase in API D-mannos, N-acetyl-glucosamine, D-maltose, gluco- 20NE but negative for nitrate reduction, indole produc- nate, caprate, adipate, malate, citrate and phenylacetate. tion, glucose fermentation, arginine dihydrolase and ure- Strain LPB0133 (=NIBRBAC000498521) has been iso- ase. D-Glucose is utilized. Does not utilize L-arabinose, lated from squid, Jumunjin, Gangwon-do, Korea. D-mannitol, D-mannose, N-acetyl-glucosamine, D-maltose, gluconate, caprate, adipate, malate, citrate and phe- Description of Kocuria carniphila PMA2305 nylacetate. Strain 5187 ( =NIBRBAC000498568) has been isolated from sediment, Han river, Korea. Cells are Gram-staining-positive, non-flagellated and coccoid shaped. Colonies are circular and cream-col- Description of Cellulosimicrobium cellulans PMA3209 ored after 2 days on MA at 20°C. Positive for nitrate reduction and urease in API 20NE but negative for in- Cells are Gram-staining-positive, non-flagellated and dole production, glucose fermentation, arginine dihydro- rod-shaped. Colonies are circular and yellow-colored lase, esculin hydrolysis, gelatinase and β-galactosidase. after 2 days on MA at 37°C. Positive for nitrate reduc- D-Glucose, D-mannitol, D-mannose, N-acetyl-glucos- tion, glucose fermentation, gelatinase and esculin hydro- amine, D-maltose, gluconate, adipate, malate, citrate and lysis in API 20NE but negative for indole production, phenylacetate are utilized. Does not utilize L-arabinose arginine dihydrolase, and β-galactosidase. D-Glucose, and caprate. Strain PMA2305 (=NIBRBAC000498398) L-arabinose, D-mannitol, D-mannose, N-acetyl-glucos- has been isolated from droppings of eagles, Seoul Grand amine, gluconate, D-maltose, adipate, malate, citrate Park, Gwacheon, Gyeonggi, Korea. and phenylacetate are utilized. Does not utilize caprate, Strain PMA3209 ( =NIBRBAC000498393) has been Description of Micrococcus lylae LPB0125 isolated from droppings of spoonbills, Seoul Grand Park, Gwacheon, Gyeonggi, Korea. Cells are Gram-staining-positive, non-flagellated, non-pigmented and rod-shaped. Colonies are circular, Description of Oerskovia turbata YC4-25 convex, entire and yellow-colored after 1 day on MA at 25°C. Positive for gelatinase in API 20NE but neg- Cells are Gram-staining-positive, non-flagellated, ative for nitrate reduction, indole production, glucose non-pigmented and rod to coccoid-shaped. Colonies are fermentation, arginine dihydrolase, urease, esculin hy- circular, convex, entire and light yellow-colored after February 2018 Lee et al. - Unrecorded actinobacterial species in Korea 47

4 days on NA at 30°C. Positive for nitrate reduction, Description of Streptomyces durhamensis Gsoil 224 esculin hydrolysis and β-galactosidase in API 20NE, but Cells are Gram-staining-positive, non-flagellated and negative for indole production, glucose fermentation, produce well-developed, branched aerial and vegetative arginine dihydrolase, urease and gelatinase. D-Glu- mycelia. Colonies are round, convex, circular, entire cose, D-mannitol and N-acetyl-glucosamine are weak- and gray-colored after 3 days on R2A at 25°C. Positive ly utilized. L-arabinose, D-maltose and gluconate are for nitrate reduction and β-galactosidase in API 20NE utilized. Does not utilize D-mannose, caprate, adipate, but negative for indole production, glucose fermenta- malate, citrate and phenylacetate. Strain YC4-25 (=NI- tion, arginine dihydrolase, urease, esculin hydrolysis BRBA000498651) has been isolated from Yongcheon and gelatinase. D-Glucose, L-arabinose, D-mannitol, Cave, Jeju, Koraea. D-mannose, N-acetyl-glucosamine, D-maltose, gluconate, adipate, malate, citrate and phenylacetate are uti- Description of Micromonospora echinospora Gsoil 1174 lized. Does not utilize caprate. Strain Gsoil 224 (=NI- Cells are Gram-staining-positive, non-flagellated and BRBAC000498585) has been isolated from ginseng substrate mycelium-forming. Colonies are convex, irreg- soil, Anseong, Korea. ulaer and brown-colored after 3 days on R2A at 25°C. Negative for nitrate reduction, indole production, glu- Description of Streptomyces kurssanovii UL451 cose fermentation, arginine dihydrolase, urease, esculin Cells are Gram-staining-positive, non-flagellated, hydrolysis, gelatinase and β-galactosidase in API 20NE. non-pigmented and produce well-developed, branched L-Arabinose, D-mannitol, gluconate, malate are utilized. aerial and vegetative mycelia. Colonies are circular, Does not utilize D-glucose, D-mannose, N-acetyl-glu- convex, rough, entire and mud yellow-colored after 3 cosamine, D-maltose, caprate, adipate, citrate and phe- days on TSA at 30°C. Positive for esculin hydrolysis = nylacetate. Strain Gsoil 1174 ( NIBRBAC000498604) and β-galactosidase in API 20NE but negative for ni- has been isolated from ginseng soil, Anseong, Korea. trate reduction, indole production, glucose fermentation, arginine dihydrolase, urease and gelatinase. D-Glucose, Description of Aeromicrobium fastidiosum YC3-14 L-arabinose, D-mannitol, D-mannose, N-acetyl-glucos- Cells are Gram-staining-positive, non-flagellated, amine, D-maltose, gluconate and malate are utilized. non-pigmented and rod-shaped. Colonies are circular, Does not utilize caprate, adipate, citrate and phenylace- = convex, filamentous and cream-colored after 5 days on tate. Strain UL451 ( NIBRBAC000498617) has been ISP2 at 30°C. Negative for nitrate reduction, indole pro- isolated from soil, Ulleung-gun, Gyeongsangbuk-do, duction, glucose fermentation, arginine dihydrolase, ure- Korea. ase, esculin hydrolysis, gelatinase and β-galactosidase in API 20NE. Does not utilize D-glucose, L-arabinose, Description of Streptomyces blastmyceticus D-mannitol, D-mannose, N-acetyl-glucosamine, D-malt- MMS16-CNU183 ose, gluconate, caprate, adipate, malate, citrate and phe- Cells are Gram-staining-positive, non-flagellated, nylacetate. Strain YC3-14 ( =NIBRBA000498650) has non-pigmented and produce well-developed, branched been isolated from Yongcheon Cave, Jeju, Korea. aerial and vegetative mycelia. Colonies are circular, convex, rough, entire and mud yellow-colored after 3 Description of Kitasatospora kifunensis days on ISP2 (pH 5) at 30°C. Positive for urease, gelati- MMS16-CNU292 nase, esculin hydrolysis and β-galactosidase in API 20NE but negative for nitrate reduction, indole produc- Cells are Gram-staining-positive, non-flagellated and tion, glucose fermentation and arginine dihydrolase. produce well-developed, branched aerial and vegetative Does not utilize D-glucose, L-arabinose, D-mannitol, mycelia. Colonies are circular, dry, rough, opaque and D-mannose, N-acetyl-glucosamine, D-maltose, gluco- beige-colored after 3 days on SCA (pH 5) at 30°C. Pos- nate, caprate, adipate, malate, citrate and phenylacetate. itive for esculin hydrolysis and gelatinase in API 20NE Strain MMS16-CNU183 ( =NIBRBAC000498623) has but negative for nitrate reduction, indole production, been isolated from soil, Daejeon, Korea. glucose fermentation and arginine dihydrolase, urease and β-galactosidase. D-Glucose is utilized. Does not uti- Description of Nocardiopsis synnemataformans lize L-arabinose, D-mannitol, D-mannose, N-acetyl-glu- LPB0126 cosamine, D-maltose, gluconate, caprate, adipate, malate, citrate and phenylacetate. Strain MMS16-CNU292 Cells are Gram-staining-positive, non-flagellated, ( =NIBRBAC000498624) has been isolated from soil, non-pigmented and produce branched aerial and veg- Daejeon, Korea. etative mycelia. Colonies are circular, convex, entire 48 Journal of Species Research Vol. 7, No. 1 and white-colored after 1 day on MA at 25°C. Positive evolution: minimum change for a specific tree topology. for nitrate reduction and β-galactosidase in API 20NE Systematic Zoology 20:406-416. but negative indole production, glucose fermentation, Hasegawa, S., A. Meguro, M. Shimizu, T. Nishimura and H. arginine dihydrolase, urease, esculin hydrolysis and Kunoh. 2006. Endophytic actinomycetes and their inter- gelatinase. D-Mannos and malate are utilized. Does actions with host plants. Actinomycetologia 20:72-81. not utilize D-glucose, L-arabinose, D-mannitol, N-ace- Jukes, T.H. and C.R. Cantor. 1969. Evolution of protein tyl-glucosamine, D-maltose, gluconate, caprate, adi- molecules. In: H.N. Munro (ed.), Mammalian Protein pate, citrate and phenylacetate. Strain LPB0126 ( =NI- Metabolism, Academic Press, New York. pp. 21-132. BRBAC000498516) has been isolated from squid, Ju- Kim, O.S., Y.J. Cho, K. Lee, S.H. Yoon, M. Kim, H. Na, S.C. munjin, Gangwon-do, Korea. Park, Y.S. Jeon, J.H. Lee, H. Yi, S. Won and J. Chun. 2012. Introducing EzTaxon-e: a prokaryotic 16S rRNA Description of Nonomuraea harbinensis Gsoil 1046 gene sequence database with phylotypes that represent uncultured species. International Journal of Systematic Cells are Gram-staining-positive, non-flagellated and and evolutionary Microbiology 62:716-721. mycelium-forming. Colonies are convex, circular and Küster, E. and S.T. Williams. 1964. Selection of media for white-colored after 3 days on R2A at 25°C. Positive for isolation of streptomycetes. Nature 202:928–929. nitrate reduction and β-galactosidase in API 20NE but Lechevalier, H.A. 1989. The actinomycetes III - A practical negative for indole production, glucose fermentation, guide to generic identification of actinomycetes. In: S.T. arginine dihydrolase, urease, esculin hydrolysis and Williams, M.E. Sharpe and J.G. Holt (eds.), Bergey’s gelatinase. D-Glucose, L-arabinose, D-mannitol, D-man- Manual of Systematic Bacteriology, vol. 4, Williams & nose, N-acetyl-glucosamine, gluconate are utilized. Wilkins, Baltimore. pp. 2344-2347. Does not utilize D-maltose, caprate, adipate, malate, ci- Lechevalier, H.A. and M.P. Lechvalier. 1981. Introduction trate and phenylacetate. Strain Gsoil 1046 (=NIBRBAC to order Actinomycetales. In: M.P. Starr, H. Stolp, H.G. 000498588) has been isolated from ginseng soil, An- Trüper, A. Balows and H.G. Schlegel (eds.), The Pro- seong, Korea. karyotes: a handbook on habitats, isolation and identification of bacteria. vol. 2, Springer, Berlin. pp. 1915- 1922. Acknowledgements Ludwig, W., J. Euzéby, P. Schumann, H.-J. Busse, M.E. Trujillo, P. Kämpfer and W.B. Whitman. 2012. Road This study was supported by the research grant “The map of the phylum Actinobacteria. In: M. Goodfellow, Survey of Korean Indigenous Species” from the Nation- P. Kämpfer, H.-J. Busse, M.E. Trujillo, K. Suzuki, W. al Institute of Biological Resources of the Ministry of Ludwig and W.B. Whitman (eds.), Bergey’s Manual of Environment in Korea. Systematic Bacteriology (2nd ed), vol. 5, Springer, New York. pp. 1-28. Mincer, T.J., P.R. Jensen, C.A. Kauffman and W. Fenical. References 2002. Widespread and persistent populations of a major new marine actinomycete taxon in ocean sediments. Ap- Busti, E., P. Monciardini, L. Cavaletti, R. Bamonte, A. La- plied and Environmental Microbiology 68:5005-5011. zzarini, A. Sosio and S. Donadio. 2006. Antibiotic-pro- Nolan, R.D. and T. Cross. 1988. Isolation and screening of ducing ability by representatives of a newly discovered actinomycetes. In: M. Goodfellow, S.T. Williams and M. lineage of actinomycetes. Microbiology 152:675-683. Modarski (eds.), Actinomycetes in Biotechnology, Aca- Coombs, J.T. and C.M.M. Franco. 2003. Isolation and iden- demic Press, London. pp. 1-32. tification of actinobacteria from surface-sterilized wheat Prescott, L.M., J.P. Harley and D.A. Klein. 2002. Bacteria: roots. Applied and Environmental Microbiology 69: The high G+C Gram positives. In Microbiology (5th ed), 5603-5608. USA, The McGraw-Hill Company. pp. 536-551. Ensign, J.C. 1992. Introduction to the actinomycetes. In: A. Saitou, N. and M. Nei. 1987. The neighbor-joining method: Balows, H.G. Trüper, M. Dworkin, W. Harder and K.H. a new method for reconstructing phylogenetic trees. Mo- Schleifer (eds.), The Prokaryotes, vol. 1, Springer Verlag, lecular Biology and Evolution 4:406-425. New York. pp. 811-815. Shirling, E.B. and D. Gottlieb. 1966. Methods for character- Felsenstein, J. 1981. Evolutionary trees from DNA sequences: ization of Streptomyces species. International Journal of a maximum likelihood approach. Journal of Molecular Systematic Bacteriology 16:313-340. Evolution 17:368-376. Thompson, J.D., T.J. Gibson, F. Plewniak, F. Jeanmougin Felsenstein, J. 1985. Confidence limits on phylogenies: an and D.G. Higgins. 1997. The ClustalX windows inter- approach using the bootstrap. Evolution 39:783-791. face: flexible strategies for multiple sequence alignment Fitch, W.M. 1971. Towards defining the course of course of aided by quality analysis tools. Nucleic Acids Research February 2018 Lee et al. - Unrecorded actinobacterial species in Korea 49

24:4876-4882. mycetes, Academic Press, London. pp. 481-528. Williams, S.T., M. Goodfellow and G. Alderson. 1989. Ge- Zhang, M.M., M. Poulsen and C.R. Currie. 2007. Symbiont nus Streptomyces Waksman and Henrici, 1943.339AL. recognition of mutualistic bacteria by Acromyrmex leaf In: S.T. Williams, M.E. Sharpe and J.G. Holt (eds.), cutter ants. ISME Journal 1:313-320. Bergey’s Manual of Systematic Bacteriology, vol. 4, Springer, New York. pp. 2452-2492. Williams, S.T., S. Lanning and E.M.H. Wellington. 1984. Submitted: September 4, 2017 Ecology of actinomycetes. In: M. Goodfellow, M. Mo- Revised: December 26, 2017 darski and S.T. Williams (eds.), The Biology of Actino- Accepted: December 29, 2017