A Report of 11 Unrecorded Bacterial Species in Korea Isolated in 2017

Journal of Species Research 7(2):135-150, 2018

A report of 11 unrecorded bacterial species in Korea isolated in 2017

Soohyun Maeng1, Ju-Young Kim2, Jun Hwee Jang2, Myung-Suk Kang3 and Myung Kyum Kim2,*

1Department of Public Health Sciences, Graduate School, Korea University, Seoul 02841, Republic of Korea 2Department of Bio & Environmental Technology, College of Natural Science, Seoul Women’s University, Seoul 01797, Republic of Korea 3Microorganism Resources Division, National Institute of Biological Resources, Incheon 22689, Republic of Korea

*Correspondent: [email protected]

Eleven bacterial strains 17SD2_15, 17Sr1_23, 17SD2_13, 17Sr1_31, 17gy_18, 16B15D, 16B02D, 16B04G, 16B01D, 17U4-2 and 17J28-10 assigned to the phylum Proteobacteria were isolated from soil samples collected from Seoul Women’s University, in South Korea. The Belnapia species, strain 17SD2_15 was cocci-shaped and pink-colored. The Methylobacterium species, strain 17Sr1_23, 17SD2_13, 17Sr1_31, and 16B15D were short rod-shaped and pink-colored. The Microvirga species, strain 17gy_18, and 16B02D were short rod-shaped and pink-colored. The Oxalicibacterium species, strain 16B04G was short rod-shaped and pink-colored. The Sphingomonas species, strain 16B01D was short rod-shaped and yellow-colored. The Variovorax species, strain 17U4-2 was cocci-shaped and yellow-colored. The Paracoccus species, 17J28-10 was cocci-shaped and orange-colored. Phylogenetic analysis based on 16S rRNA gene sequence showed that strains 17SD2_15, 17Sr1_23, 17SD2_13, 17Sr1_31, 17gy_18, 16B15D, 16B02D, 16B04G, 16B01D, 17U4- 2 and 17J28-10 were most closely related to Belnapia soli (with 99.9% similarity), Methylobacterium gregans (99.1%), Methylobacterium isbiliense (99.6%), Methylobacterium oxalidis (99.9%), Microvirga aerilata (98.7%), Methylobacterium aerolatum (99.0%), Microvirga vignae (100.0%), Noviherbaspirillum canariense (100.0%), Sphingomonas desiccabilis (100.0%), Variovorax humicola (99.6%), and Paracoccus acridae (99.1%), respectively. This is the first report of these eleven species in Korea. Keywords: 16S rRNA, bacterial diversity, Proteobacteria, unreported species

Introduction of energy and carbon (Gallego et al., 2005; Konovalova et al., 2007; Cao et al., 2011). The family Methylobacteri- In 2017, eleven unreported species were isolated from aceae are pink-pigmented and contain ubiquinone Q-10. diverse soil samples collected in Korea. The current re- The eight species of Microvirga and the single species port focuses on the description of eleven unreported bac- of Meganema are not methylotrophic (Kanso and Patel terial species belonging to phylum Proteobacteria that 2003; Weon et al., 2010; Ardley et al., 2012). have not officially reported in Korea. Remaining five Strains 17SD2_15, 16B04G, 16B01D, Six bacterial strains 17Sr1_23, 17SD2_13, 17Sr1_31, 17U4-2 and 17J28-10 belonged to the family Acetobacte- 17gy_18, 16B15D and 16B02D belong to the family raceae, Oxalobacteraceae, Sphingomonadaceae, Coma- Methylobacteriaceae. The family Methylobacteriaceae is monadaceae, and Rhodobacteraceae, respectively. Ace- one of the large family of Alphaproteobacteria (Garrity tobacteraceae are included in the order Rhodospirillales et al., 2005a; Kelly et al., 2014) and at present contains and 32 genera are validly published (http://www.bacterio. three genera, Methylobacterium, Microvirga and Megane- net/-classifphyla.html#rhodospirillales). The genera in ma (http://www.bacterio.net/). The genus Methylobacte- the family Acetobacteraceae are basically classified into rium is one of the largest genus containing 44 validated two groups, an acetous group and an acidophilic group, species. They are facultative methylotrophs able to grow in the light of application, ecology, and phylogeny on methanol and other one-carbon compounds as sources (Komagata et al., 2014). 136 JOURNAL OF SPECIES RESEARCH Vol. 7, No. 2

Table 1. 16S rRNA gene sequence similarity, isolation source, medium, and incubation conditions of unrecorded strains. Strain ID Most closely related species Similarity (%) Isolation source Medium Incubation conditions 17SD2_15 Belnapia soli 99.9 Soil R2A 25°C, 3d 17Sr1_23 Methylobacterium gregans 99.1 Soil R2A 25°C, 3d 17SD2_13 Methylobacterium isbiliense 99.6 Soil R2A 25°C, 3d 17Sr1_31 Methylobacterium oxalidis 99.9 Soil R2A 25°C, 3d 17gy_18 Microvirga aerilata 98.7 Soil R2A 25°C, 3d 16B15D Methylobacterium aerolatum 99.0 Soil R2A 25°C, 3d 16B02D Microvirga vignae 100.0 Soil R2A 25°C, 3d 16B04G Oxalicibacterium flavum 100.0 Soil R2A 25°C, 3d 16B01D Sphingomonas desiccabilis 100.0 Soil R2A 25°C, 3d 17U4-2 Variovorax humicola 99.6 Soil R2A 25°C, 3d 17J28-10 Paracoccus acridae 99.1 Soil R2A 25°C, 3d

The family Oxalobacteraceae which contains the 13 in distilled water and serially diluted. The 100 μL of the genera (Xu et al., 2005) is belonged to the order Burk- aliquot was inoculated onto R2A agar and incubated at holderiales. The family Sphingomonadaceae was pro- 25°C for 3 days. The designated strain IDs, isolation posed based on the 16S rRNA gene sequence phylogeny sources, growth media, and incubation conditions are and the presence of 2ʹhydroxymyristol dihydrosphin- summarized in Table 1. All strains were purified as sin- gosine 1-glucuronic acid (SGL-1) as major sphingogly- gle colonies and stored as 20% glycerol suspension at colipid in the cellular lipids (Kosako et al., 2000). Sphin- -80°C as well as freeze-dried ampoules. gomonadaceae is belonged to the order Sphingomonad- The colony morphology and cell size of the strains ales which contains the 18 genera at the time of writing were observed by transmission electron microscopy (LI (http://www.bacterio.net/-classifphyla.html#sphingo- BRA 120, Carl Zeiss) using cells grown for 3 days at monadaceae). The Sphingomonadaceae family was pro- 25°C on R2A agar. Transmission electron micrograph of posed based on the 16S rRNA gene sequence phylogeny the strains are shown in Fig. 1. Gram reaction was per- and the presence of 2ʹhydroxymyristol dihydrosphin- formed following the classic Gram procedure described gosine 1-glucuronic acid (SGL-1) as major sphingogly- by Doetsch (1981). Biochemical characteristics were test- colipid in the cellular lipids (Kosako et al. 2000). ed by using Biolog Microstation with GEN III microplate The family Comamonadaceae is one of the biggest system. A single colony was selected and emulsified into family belonging to the order Burkholderiales in which ‘inoculating fluid A’ (Biolog) for subsequent inoculation contains 37 genera (http://www.bacterio.net/-classifphy- on to the Microplate test plate (Biolog). More fastidious la.html#comamonadaceae). These genera form a phylo- organisms, including capnophilic strains, were cultured genetic cluster based on 16S rRNA gene sequence sim- on alternative media, according to the manufacturer’s ilarity of 93-97% yet, have a diverse phenotypic char- instructions, and inocula were prepared to a specified acteristic that includes aerobic organotrophs, anaerobic transmittance using a turbidimeter, as specified in the + denitrifiers, Fe3 -reducing bacteria, hydrogen oxidizers, user guide. For each isolate, 100 μL of the cell suspen- photoautotrophic and photoheterotrophic bacteria, and sion was inoculated into each well of the microplate, us- fermentative bacteria (Willems et al., 1991). ing a multichannel pipette and incubated at 37°C for 24 The family Rhodobacteraceae are basically, aquatic h, according to growth characteristics. Microplates were bacteria that frequently thrive in marine environments. read in the MicroStation semi-automated reader after 24 They include mainly aerobic photo- and chemohetero- h and results interpreted by the identification system’s trophs but also purple non-sulfur bacteria which perform software (GEN III database, version 5.2.1). Whereas photosynthesis in anaerobic environments. They are the system which indicates that the isolates could not deeply involved in sulfur and carbon biogeochemical be identified after 20 h and required further incubation. cycling and symbiosis with aquatic micro- and macroor- Such isolates were re-incubated and re-read between 3 ganisms (Garrity et al., 2005b). and 6 h later (Wragg et al., 2014). Genomic DNA was extracted and 16S rRNA gene was amplified by PCR with 9F and 1492R universal bacterial primers (Weisburg Material and Methods et al., 1991). The 16SrRNA gene sequences of the closely related strains were obtained from EzTaxon-e (http:// Different soil samples were collected and suspended eztaxon-e.ezbiocloud.net) (Kim et al., 2012) and edited May 2018 Maeng et al. Report of 11 unrecorded radiation resistant bacteria 137

(1) (2) (3) (4)

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Fig. 1. Transmission electron micrographs of the strains isolated in this study. Strains: 1, 17SD2_15; 2, 17Sr1_23; 3, 17SD2_13; 4, 17Sr1_31; 5, 17gy_18; 6, 16B15D; 7, 16B02D; 8, 16B04G; 9, 16B01D; 10, 17U4-2; 11, 17J28-10. using the BioEdit program (Hall, 1999). Multiple align- different genera: Belnapia (1 species), Methylobacteri- ments were performed with the MUSCL program (Edgar, um (4 species), Microvirga (2 species), Sphingomonas 2004). Using the two-parameter model (Kimura, 1983) (1 species), Oxalicibacterium (1 species), Variovorax (1 calculated the evolutionary distances. Phylogenetic trees species), and Paracoccus (1 species). The identification were constructed using the neighbor-joining (Saitou and of the isolates based on sequence similarity was sup- Nei, 1987) in the MEGA5 program (Tamura, 2011) with ported by the phylogenetic trees. The neighbor-joining bootstrap values based on 1,000 replications (Felsenstein, trees show the close relationship of the isolates and type 1985). strains of validly published species (Fig. 1). The detailed morphological and physiological characteristics are giv- en in the strain descriptions. esults and iscussion R D As an outcome of this study, the diversity of radia- Based on 16S rRNA gene sequence similarity, 11 of tion-resistant bacterial species is not reported previously previously unreported bacterial species were identified. in Korean ecosystems was discovered. The 11 isolates The taxonomic composition and identification results were identified as unreported species, and their pheno- are summarized in Table 1. The 11 strains were assigned typic characteristics were examined through polyphasic to the family Acetobacteraceae (1 strain), Methylobac- study. Accordingly, the following 11 species are unre- teriaceae (6 strain), Oxalobacteraceae (1 strain), and ported species in Korea. Sphingomonadaceae (1 strain), Comamonadaceae (1 strain), Rhodobacteraceae (1 strain) of the phylum Pro- Description of Belnapia soli 17SD2_15 teobacteria. At the generic level, the strains belong to 7 Cells are Gram-stain-negative, non-flagellated, and 138 JOURNAL OF SPECIES RESEARCH Vol. 7, No. 2

Fig. 2. Neighbor-joining phylogenetic tree based on 16S rRNA gene sequences shows the relationship between the strains isolated in this study and their relatives of the genus Belnapia. Bootstrap values (>70%) are shown above nodes for the neighbor-joining methods. Bar: 0.01 and 0.02 substitutions per nucleotide position, respectively.

Fig. 3. Neighbor-joining phylogenetic tree based on 16S rRNA gene sequences shows the relationship between the strains isolated in this study and their relatives of the genus Methylobacterium. Bootstrap values (>70%) are shown above nodes for the neighbor-joining methods. Bar: 0.01 and 0.02 substitutions per nucleotide position, respectively. coccus-shaped. Colonies are pale pinkish-colored after acid, N-acetyl-D-galactosamine, N-acetyl-neuraminic 3 days of incubation on R2A at 25°C. In the BIOLOG acid, N-acetyl-D-glucosamine, γ-amino-butryric acid, GEN III, D-glucuronic acid was utilized as a sole carbon D-arabitol, D-aspartic acid, bromo-succinic acid, cit- source. But acetoacetic acid, N-acetyl-D-mannosamine, ric acid, formic acid, D-fucose, L-fucose, D-glucose-6- L-alanine, L-arginine, L-aspartic acid, D-cellobiose, PO4, glycerol, L-histidine, α-hydroxybutyric acid, β-hy- dextrin, D-fructose, D-fructose 6-PO4, L-galactonic acid droxy-D, L-butyric acid, p-hydroxy-phenylacetic acid, lactone, D-galactose, D-galacturonic acid, gelatin, gen- α-keto-butyric acid, L-lactic acid, D-lactic acid methyl tiobiose, D-gluconic acid, α-D-glucose, glucuronamide, ester, α-D-lactose, D-malic acid, methyl pyruvate, mu- L-glutamic acid, inosine, α-keto-glutaric acid, L-malic cic acid, D-saccharic acid, D-salicin, D-serine, D-sorbi- acid, D-maltose, D-mannitol, D-mannose, D-melibiose, tol, and tween 40 were not utilized. In sensitivity tests, β-methyl-D-glucoside, 3-methyl glucose, myo-inositol, the tetrazolium redox dye was reduced at pH 6 but not pectin, propionic acid, glycyl-L-proline, L-pyroglutam- at 1% NaCl, 1% sodium lactate, 4% NaCl, 8% NaCl, ic acid, quinic acid, D-raffinose, L-rhamnose, L-ser- guanidine HCl, lithium chloride, pH 6, potassium tellu- ine, stachyose, sucrose, D-trehalose, D-turanose, acetic rite, D-serine, sodium butyrate, aztreonam, fusidic acid, May 2018 Maeng et al. Report of 11 unrecorded radiation resistant bacteria 139

(1)

(2)

Fig. 4. Neighbor-joining phylogenetic tree based on 16S rRNA gene sequences shows the relationship between the strains isolated in this study and their relatives of the genus Methylobacterium (1), (2). Bootstrap values (>70%) are shown above nodes for the neighbor-joining methods. Bar: 0.01 and 0.02 substitutions per nucleotide position, respectively. lincomycin, minocycline, nalidixic acid, niaproof 4, pH amine, D-ribose, inositol, D-saccharose (sucrose), D-malt- 5, rifamycin SV, sodium bromate, tetrazolium blue, tro- ose, L-alanine, potassium 5-ketogluconate, glycogen, leandomycin, and vancomycin. 3-hydroxybenzoic acid, D-mannitol, D-glucose, salicin, In API 20NE system, positive for arginine dihydrolase D-melibiose, D-fucose, D-sorbitol, L-arabinose, valeric and urease. Negative for nitrate reduction, indole produc- acid, trisodium citrate, and L-histidine. Strain 17SD2_15 tion, glucose fermentation, esculin hydrolysis, gelatin hy- (=NIBRBAC000500510) was isolated from a soil sam- drolysis, and β-galactosidase. Uses D-glucose, potassium ple. gluconate, adipic acid, malic acid, and trisodium citrate but not L-arabinose, D-mannose, D-mannitol, N-ace- Description of Methylobacterium gregans 17Sr1_23 tyl-glucosamine, capric acid, D-maltose, and phenyl acetic acid as carbon sources. Cells are Gram-stain-negative, non-flagellated, and In API 32GN system, positive for itaconic acid, sub- short rod-shaped. Colonies are pink-colored after 3 days eric acid, sodium malonate, sodium acetate, lactic acid, of incubation on R2A at 25°C. In the BIOLOG GEN III, L-serine, propionic acid, capric acid, potassium 2-keto- D-glucuronic acid and formic acid is utilized as a sole gluconate, 3-hydroxybutyric acid, 4-hydroxybenzoic acid, carbon source. But acetoacetic acid, N-acetyl-D-man- and L-proline. Negative for L-rhamnose, N-acetyl-glucos- nosamine, L-alanine, L-arginine, L-aspartic acid, D-cel- 140 JOURNAL OF SPECIES RESEARCH Vol. 7, No. 2

Fig. 5. Neighbor-joining phylogenetic tree based on 16S rRNA gene sequences shows the relationship between the strains isolated in this study and their relatives of the genus Microvirga. Bootstrap values (>70%) are shown above nodes for the neighbor-joining methods. Bar: 0.01 and 0.02 substitutions per nucleotide position, respectively.

lobiose, dextrin, D-fructose, D-fructose 6-PO4, L-ga- fermentation, gelatin hydrolysis, D-glucose, L-arabinose, lactonic acid lactone, D-galactose, D-galacturonic acid, D-mannitol, N-acetyl-D-glucosamine, potassium gluco- gelatin, gentiobiose, D-gluconic acid, α-D-glucose, glu- nate, capric acid, adipic acid, malic acid, trisodium citrate, curonamide, L-glutamic acid, inosine, α-keto-glutaric and phenylacetic acid. acid, L-malic acid, D-maltose, D-mannitol, D-mannose, In API 32GN system, positive for itaconic acid, suber- D-melibiose, β-methyl-D-glucoside, 3-methyl glucose, ic acid, lactic acid, L-alanine, propionic acid, trisodium myo-inositol, pectin, propionic acid, glycyl-L-proline, citrate, and 3-hydroxybutyric acid. Negative for L-rham- L-pyroglutamic acid, quinic acid, D-raffinose, L-rhamnose, N-acetyl-glucosamine, D-ribose, inositol, D-sac- nose, L-serine, stachyose, sucrose, D-trehalose, D-tur- charose (sucrose), D-maltose, sodium malonate, sodium anose acetic acid, N-acetyl-D-galactosamine, N-ace- acetate, potassium 5-ketogluconate, glycogen, 3-hy- tyl-neuraminic acid, N-acetyl-D-glucosamine, γ-ami- droxybenzoic acid, L-serine, D-mannitol, D-glucose, no-butryric acid, D-arabitol, D-aspartic acid, bromo-suc- salicin, D-melibiose, D-fucose, D-sorbitol, L-arabinose, cinic acid, citric acid, D-fucose, L-fucose, D-gluco se-6- capric acid, valeric acid, L-histidine, potassium 2-keto- PO4, glycerol, L-histidine, α-hydroxybutyric acid, β-hy- gluconate, 4-hydroxybenzoic acid, and L-proline. Strain droxy-D,L-butyric acid, p-hydroxy-phenylacetic acid, 17Sr1_23 ( =NIBRBAC000500515) was isolated from α-keto-butyric acid, L-lactic acid, D-lactic acid methyl a soil sample. ester, α-D-lactose, D-malic acid, methyl pyruvate, mucic acid, D-saccharic acid, D-salicin, D-serine, D-sorbi- Description of Methylobacterium isbiliense 17SD2_13 tol, and tween 40 were not utilized. In sensitivity tests, the tetrazolium redox dye is reduced in the presence of Cells are Gram-stain-negative, non-flagellated, and lincomycin, vancomycin, tetrazolium blue, nalidixic short rod-shaped. Colonies are pink-colored after 3 days acid, potassium tellurite, and aztreonam; but not in the of incubation on R2A at 25°C. In the BIOLOG GEN presence of 1% NaCl, 1% sodium lactate, 4% NaCl, 8% III acetoacetic acid, N-acetyl-D-mannosamine, L-ala- NaCl, guanidine HCl, lithium chloride, pH 6, D-serine, nine, L-arginine, L-aspartic acid, D-cellobiose, dextrin, sodium butyrate, tetrazolium violet fusidic acid, minocy- D-fructose, D-fructose 6-PO4, L-galactonic acid lactone, cline, niaproof 4, pH 5, rifamycin SV, sodium bromate, D-galactose, D-galacturonic acid, gelatin, gentiobiose, and troleandomycin. D-gluconic acid, α-D-glucose, glucuronamide, D-glu- In API 20NE system, positive for the reduction of ni- curonic acid, L-glutamic acid, inosine, α-keto-glutaric trates (NO3) to nitrogen (N2), arginine dihydrolase, ure- acid, L-malic acid, D-maltose, D-mannitol, D-mannose, ase, esculin hydrolysis, β-galactosidase, D-mannose, and D-melibiose, β-methyl-D-glucoside, 3-methyl glucose,

D-maltose. Negative for the reduction of nitrates (NO3) to myo-inositol, pectin, propionic acid, glycyl-L-proline, - nitrite (NO2 ), indole production on tryptophan, glucose L-pyroglutamic acid, quinic acid, D-raffinose, L-rham- May 2018 Maeng et al. Report of 11 unrecorded radiation resistant bacteria 141

Fig. 6. Neighbor-joining phylogenetic tree based on 16S rRNA gene sequences shows the relationship between the strains isolated in this study and their relatives of the genus Methylobacterium. Bootstrap values (>70%) are shown above nodes for the neighbor-joining methods. Bar: 0.01 and 0.02 substitutions per nucleotide position, respectively. nose, L-serine, stachyose, sucrose, D-trehalose, D-tur- ic acid, niaproof 4, pH 5, rifamycin SV, sodium bromate, anose, acetic acid, N-acetyl-D-galactosamine, N-ace- troleandomycin, and vancomycin. tyl-neuraminic acid, N-acetyl-D-glucosamine, γ-ami- In API 20NE system, positive for the reduction of - no-butryric acid, D-arabitol, D-aspartic acid, bromo-suc- nitrates (NO3) to nitrite (NO2 ), arginine dihydrolase, cinic acid, citric acid, formic acid, D-fucose, L-fucose, urease, esculin hydrolysis, β-galactosidase, D-glucose, D-glucose-6-PO4, glycerol, L-histidine, α-hydroxybu- L-arabinose, D-mannose, D-mannitol, N-acetyl-D-glu- tyric acid, β-hydroxy-D, L-butyric acid, p-hydroxy-phe- cosamine, D-maltose, potassium gluconate, adipic acid, nylacetic acid, α-keto-butyric acid, L-lactic acid, D-lac- malic acid, trisodium citrate, and phenylacetic acid. tic acid methyl ester, α-D-lactose, D-malic acid, meth- Weak positive for gelatin hydrolysis. Negative for the yl pyruvate, mucic acid, D-saccharic acid, D-salicin, reduction of nitrates (NO3) to nitrogen (N2), indole pro- D-serine, D-sorbitol, and tween 40 were not utilized. In duction on tryptophan, glucose fermentation, and capric sensitivity tests, the tetrazolium redox dye is reduced in acid. the presence of tetrazolium violet, tetrazolium blue, and In API 32GN system, positive for L-rhamnose, D-ri- aztreonam; but not in the presence of 1% NaCl, 1% so- bose, itaconic acid, suberic acid, sodium malonate, sodium lactate, 4% NaCl, 8% NaCl, guanidine HCl, lithi- dium acetate, lactic acid, potassium 2-ketogluconate, um chloride, pH 6, potassium tellurite, D-serine, sodium 3-hydroxybutyric acid, L-serine, D-glucose, L-arabinose, butyrate, fusidic acid, lincomycin, minocycline, nalidix- propionic acid, capric acid, valeric acid, trisodium citrate, 142 JOURNAL OF SPECIES RESEARCH Vol. 7, No. 2

Fig. 7. Neighbor-joining phylogenetic tree based on 16S rRNA gene sequences shows the relationship between the strains isolated in this study and their relatives of the genus Microvirga. Bootstrap values (>70%) are shown above nodes for the neighbor-joining methods. Bar: 0.01 and 0.02 substitutions per nucleotide position, respectively.

4-hydroxybenzoic acid, L-proline, and 3-hydroxyben- droxy-D, L-butyric acid, p-hydroxy-phenylacetic acid, zoic acid. Negative for N-acetyl-glucosamine, inositol, α-keto-butyric acid, L-lactic acid, D-lactic acid methyl D-saccharose (sucrose), D-maltose, L-alanine, potassium ester, α-D-lactose, D-malic acid, methyl pyruvate, mu- 5-ketogluconate, glycogen, L-serine, D-mannitol, salicin, cic acid, D-saccharic acid, D-salicin, D-serine, D-sorbi- D-melibiose, D-fucose, D-sorbitol, and L-histidine. Strain tol, and tween 40 were not utilized. In sensitivity tests, 17SD2_13 (=NIBRBAC000500516) was isolated from a the tetrazolium redox dye is reduced in the presence of soil sample. lincomycin, tetrazolium blue, aztreonam; but not in the presence of 1% NaCl, 1% sodium lactate, 4% NaCl, 8% Description of Methylobacterium oxalidis 17Sr1_31 NaCl, guanidine HCl, lithium chloride, pH 6, potassium tellurite, D-serine, sodium butyrate, tetrazolium violet Cells are Gram-stain-negative, non-flagellated, and fusidic acid, minocycline, nalidixic acid, niaproof 4, pH short rod-shaped. Colonies are orange-colored after 3 days 5, rifamycin SV, sodium bromate, troleandomycin, and of incubation on R2A at 25°C. In the BIOLOG GEN III, vancomycin. L-malic acid and acetic acid is utilized as a sole carbon In API 20NE system, positive for the reduction of - source. But acetoacetic acid, N-acetyl-D-mannosamine, nitrates (NO3) to nitrite (NO2 ), arginine dihydrolase, L-alanine, L-arginine, L-aspartic acid, D-cellobiose, urease, esculin hydrolysis, gelatin hydrolysis, β-galacto- dextrin, D-fructose, D-fructose 6-PO4, L-galactonic acid sidase, D-glucose, L-arabinose, D-mannose, D-mannitol lactone, D-galactose, D-galacturonic acid, gelatin, gen- N-acetyl-D-glucosamine, D-maltose, potassium gluco- tiobiose, D-gluconic acid, α-D-glucose, glucuronamide, nate, adipic acid, malic acid, and trisodium citrate. Weak D-glucuronic acid, L-glutamic acid, inosine, α-ke- positive for phenylacetic acid. Negative for the reduc- to-glutaric acid, D-maltose, D-mannitol, D-mannose, tion of nitrates (NO3) to nitrogen (N2), indole production D-melibiose, β-methyl-D-glucoside, 3-methyl glucose, on tryptophan, glucose fermentation, and capric acid. myo-inositol, pectin, propionic acid, glycyl-L-proline, In API 32GN system, positive for D-mannitol, D-glu- L-pyroglutamic acid, quinic acid, D-raffinose, L-rham- cose, D-sorbitol, valeric acid, and 3-hydroxybutyric acid. nose, L-serine, stachyose, sucrose, D-trehalose, D-tur- Negative for L-rhamnose, N-acetyl-glucosamine, D-ri- anose, N-acetyl-D-galactosamine, N-acetyl-neuraminic bose, inositol, D-saccharose (sucrose), D-maltose, itacon- acid, N-acetyl-D-glucosamine, γ-amino-butryric acid, ic acid, suberic acid, sodium malonate, sodium acetate, D-arabitol, D-aspartic acid, bromo-succinic acid, cit- lactic acid, L-alanine, potassium 5-ketogluconate, glyco- ric acid, formic acid, D-fucose, L-fucose, D-glucose-6- gen, 3-hydroxybenzoic acid, L-serine, salicin, D-melibi- PO4, glycerol, L-histidine, α-hydroxybutyric acid, β-hy- ose, D-fucose, L-arabinose, propionic acid, capric acid, May 2018 Maeng et al. Report of 11 unrecorded radiation resistant bacteria 143

Fig. 8. Neighbor-joining phylogenetic tree based on 16S rRNA gene sequences shows the relationship between the strains isolated in this study and their relatives of the genus Noviherbaspirillum. Bootstrap values (>70%) are shown above nodes for the neighbor-joining methods. Bar: 0.01 and 0.02 substitutions per nucleotide position, respectively. valeric acid, trisodium citrate, L-histidine, potassium D-galactose, D-galacturonic acid, gelatin, gentiobiose, 2-ketogluconate, 3-hydroxybutyric acid, L-serine, 4-hy- D-gluconic acid, α-D-glucose, glucuronamide, D-glu- droxybenzoic acid, and L-proline. Strain 17Sr1_31 (=NI curonic acid, L-glutamic acid, inosine, α-keto-glutaric BRBAC000500517) was isolated from a soil sample. acid, L-malic acid, D-maltose, D-mannitol, D-mannose, D-melibiose, β-methyl-D-glucoside, 3-methyl glucose, Description of Microvirga aerilata 17gy_18 myo-inositol, pectin, propionic acid, glycyl-L-proline, L-pyroglutamic acid, quinic acid, D-raffinose, L-rham- Cells are Gram-stain-negative, non-flagellated, and nose, L-serine, stachyose, sucrose, D-trehalose, D-tur- short rod-shaped. Colonies are pink-colored after 3 days anose acetic acid, N-acetyl-D-galactosamine, N-ace- of incubation on R2A at 25°C. In the BIOLOG GEN tyl-neuraminic acid, N-acetyl-D-glucosamine, γ-ami- III, acetoacetic acid, N-acetyl-D-mannosamine, L-ala- no-butryric acid, D-arabitol, D-aspartic acid, bromo-suc- nine, L-arginine, L-aspartic acid, D-cellobiose, dextrin, cinic acid, citric acid, formic acid, D-fucose, L-fucose, D-fructose, D-fructose 6-PO4, L-galactonic acid lactone, D-glucose-6-PO4, glycerol, L-histidine, α-hydroxybu- 144 JOURNAL OF SPECIES RESEARCH Vol. 7, No. 2

Fig. 9. Neighbor-joining phylogenetic tree based on 16S rRNA gene sequences shows the relationship between the strains isolated in this study and their relatives of the genus Sphingomonas. Bootstrap values (>70%) are shown above nodes for the neighbor-joining methods. Bar: 0.01 and 0.02 substitutions per nucleotide position, respectively. tyric acid, β-hydroxy-D, L-butyric acid, p-hydroxy-phe- duction on tryptophan, glucose fermentation, arginine nylacetic acid, α-keto-butyric acid, L-lactic acid, D-lac- dihydrolase, gelatin hydrolysis, β-galactosidase, D-man- tic acid methyl ester, α-D-lactose, D-malic acid, meth- nitol, D-maltose, potassium gluconate, capric acid, ma- yl pyruvate, mucic acid, D-saccharic acid, D-salicin, lic acid, trisodium citrate. D-serine, D-sorbitol, and tween 40 were not utilized. In In API 32GN system, positive for N-acetyl-glucos- sensitivity tests, the tetrazolium redox dye is reduced in amine, D-saccharose (sucrose), itaconic acid, sodium the presence of tetrazolium violet; but not in the pres- malonate, lactic acid, D-glucose, D-fucose, and L-arab- ence of 1% NaCl, 1% sodium lactate, 4% NaCl, 8% inose. Negative for L-rhamnose, D-ribose, inositol, NaCl, guanidine HCl, lithium chloride, pH 6, potassium D-maltose, suberic acid, L-alanine, potassium 5-keto- tellurite, D-serine, sodium butyrate, aztreonam, fusidic gluconate, glycogen, 3-hydroxybenzoic acid, L-serine, acid, lincomycin, minocycline, nalidixic acid, niaproof 4, D-mannitol, sodium acetate, salicin, D-melibiose, D-sor- pH 5, rifamycin SV, sodium bromate, tetrazolium blue, bitol, propionic acid, capric acid, valeric acid, trisodium troleandomycin, and vancomycin. citrate, L-histidine, 4-hydroxybenzoic acid, and L-pro- In API 20NE system, positive for the reduction of ni- line. Strain 17gy_18 (=NIBRBAC000500518) was iso- - trates (NO3) to nitrite (NO2 ), urease, esculin hydrolysis, lated from a soil sample. D-glucose, L-arabinose, D-mannose, N-acetyl-D-glucosamine, adipic acid, phenylacetic acid, potassium 2-keto- Description of Methylobacterium aerolatum 16B15D gluconate, and 3-hydroxybutyric acid. Negative for the reduction of nitrates (NO3) to nitrogen (N2), indole pro- Cells are Gram-stain-negative, non-flagellated, and May 2018 Maeng et al. Report of 11 unrecorded radiation resistant bacteria 145

Fig. 10. Neighbor-joining phylogenetic tree based on 16S rRNA gene sequences shows the relationship between the strains isolated in this study and their relatives of the genus Variovorax. Bootstrap values (>70%) are shown above nodes for the neighbor-joining methods. Bar: 0.01 and 0.02 substitutions per nucleotide position, respectively. short rod-shaped. Colonies are pink-colored after 3 days acid, methyl pyruvate, mucic acid, D-saccharic acid, of incubation on R2A at 25°C. In the BIOLOG GEN D-salicin, D-serine, D-sorbitol, and tween 40 were not III, acetoacetic acid, N-acetyl-D-mannosamine, L-ala- utilized. In sensitivity tests, the tetrazolium redox dye is nine, L-arginine, L-aspartic acid, D-cellobiose, dextrin, reduced in the presence tetrazolium violet; but not in the D-fructose, D-fructose 6-PO4, L-galactonic acid lactone, presence of 1% NaCl, 1% sodium lactate, 4% NaCl, 8% D-galactose, D-galacturonic acid, gelatin, gentiobiose, NaCl, guanidine HCl, lithium chloride, pH 6, potassium D-gluconic acid, α-D-glucose, glucuronamide, D-glu- tellurite, D-serine, sodium butyrate, aztreonam, fusidic curonic acid, L-glutamic acid, inosine, α-keto-glutaric acid, lincomycin, minocycline, nalidixic acid, niaproof 4, acid, L-malic acid, D-maltose, D-mannitol, D-mannose, pH 5, rifamycin SV, sodium bromate, tetrazolium blue, D-melibiose, β-methyl-D-glucoside, 3-methyl glucose, troleandomycin, and vancomycin. myo-inositol, pectin, propionic acid, glycyl-L-pro- In API 20NE system, positive for the reduction of ni- - line, L-pyroglutamic acid, quinic acid, D-raffinose, trates (NO3) to nitrite (NO2 ), arginine dihydrolase, ure- L-rhamnose, L-serine, stachyose, sucrose, D-trehalose, ase, D-glucose, L-arabinose, D-mannose, D-mannitol, D-turanose acetic acid, N-acetyl-D-galactosamine, D-maltose, potassium gluconate, adipic acid, and malic N-acetyl-neuraminic acid, N-acetyl-D-glucosamine, acid. Weak positive for trisodium citrate. Negative for

γ-amino-butryric acid, D-arabitol, D-aspartic acid, bro- the reduction of nitrates (NO3) to nitrogen (N2), indole mo-succinic acid, citric acid, formic acid, D-fucose, production on tryptophan, glucose fermentation, esculin L-fucose, D-glucose-6-PO4, glycerol, L-histidine, α-hy- hydrolysis, gelatin hydrolysis, β-galactosidase, N-acetyl- droxybutyric acid, β-hydroxy-D, L-butyric acid, p-hy- D-glucosamine, capric acid, and phenylacetic acid. droxy-phenylacetic acid, α-keto-butyric acid, L-lactic In API 32GN system, positive for D-ribose, itaconic acid, D-lactic acid methyl ester, α-D-lactose, D-malic acid, D-glucose, and L-arabinose. Negative for L-rham- 146 JOURNAL OF SPECIES RESEARCH Vol. 7, No. 2

Fig. 11. Neighbor-joining phylogenetic tree based on 16S rRNA gene sequences shows the relationship between the strains isolated in this study and their relatives of the genus Paracoccus. Bootstrap values (>70%) are shown above nodes for the neighbor-joining methods. Bar: 0.01 and 0.02 substitutions per nucleotide position, respectively. nose, inositol, N-acetyl-glucosamine, D-saccharose D-galactose, D-galacturonic acid, gelatin, gentiobiose, (sucrose), D-maltose, suberic acid, sodium malonate, D-gluconic acid, α-D-glucose, glucuronamide, D-glu- sodium acetate, lactic acid, L-alanine, potassium 5-keto- curonic acid, L-glutamic acid, inosine, α-keto-glutaric gluconate, glycogen, 3-hydroxybenzoic acid, L-serine, acid, L-malic acid, D-maltose, D-mannitol, D-mannose, D-mannitol, salicin, D-melibiose, D-fucose, D-sorbitol, D-melibiose, β-methyl-D-glucoside, 3-methyl glucose, propionic acid, capric acid, valeric acid, trisodium ci- myo-inositol, pectin, propionic acid, glycyl-L-pro- trate, L-histidine, potassium 2-ketogluconate, 3-hydro line, L-pyroglutamic acid, quinic acid, D-raffinose, xybutyric acid, L-serine, 4-hydroxybenzoic acid, and L- L-rhamnose, L-serine, stachyose, sucrose, D-trehalose, proline. Strain 16B15D ( =NIBRBAC000500519) was D-turanose acetic acid, N-acetyl-D-galactosamine, isolated from a soil sample. N-acetyl-neuraminic acid, N-acetyl-D-glucosamine, γ-amino-butryric acid, D-arabitol, D-aspartic acid, bro- Description of Microvirga vignae 16B02D mo-succinic acid, citric acid, formic acid, D-fucose, L-fucose, D-glucose-6-PO4, glycerol, L-histidine, α-hy- Cells are Gram-stain-negative, non-flagellated, and droxybutyric acid, β-hydroxy-D, L-butyric acid, p-hy- short rod-shaped. Colonies are pink-colored after 3 days droxy-phenylacetic acid, α-keto-butyric acid, L-lactic of incubation on R2A at 25°C. In the BIOLOG GEN acid, D-lactic acid methyl ester, α-D-lactose, D-malic III, acetoacetic acid, N-acetyl-D-mannosamine, L-ala- acid, methyl pyruvate, mucic acid, D-saccharic acid, nine, L-arginine, L-aspartic acid, D-cellobiose, dextrin, D-salicin, D-serine, D-sorbitol, and tween 40 were not D-fructose, D-fructose 6-PO4, L-galactonic acid lactone, utilized. In sensitivity tests, the tetrazolium redox dye is May 2018 Maeng et al. Report of 11 unrecorded radiation resistant bacteria 147 reduced in the presence tetrazolium violet; but not in the glucuronamide, and quinic acid were not utilized. In presence of 1% NaCl, 1% sodium lactate, 4% NaCl, 8% sensitivity tests, the tetrazolium redox dye is reduced in NaCl, guanidine HCl, lithium chloride, pH 6, potassium the presence of 1% NaCl, 1% sodium lactate, 4% NaCl, tellurite, D-serine, sodium butyrate, aztreonam, fusidic lithium chloride, pH 6, potassium tellurite, D-serine, acid, lincomycin, minocycline, nalidixic acid, niaproof 4, sodium butyrate, aztreonam, nalidixic acid, niaproof 4, pH 5, rifamycin SV, sodium bromate, tetrazolium blue, sodium bromate; but not in the presence of pH 5, 8% troleandomycin, and vancomycin. NaCl, fusidic acid, troleandomycin, rifamycin SV, mino- In API 20NE system, positive for arginine dihydrolase, cycline, gelatin, lincomycin, guanidine HCl, vancomy- urease, β-galactosidase, D-glucose, L-arabinose, D-man- cin tetrazolium violet, tetrazolium blue. nose, malic acid, and trisodium citrate. Weak positive for In API 20NE system, positive for arginine dihydro-

D-mannitol. Negative for the reduction of nitrates (NO3) lase, urease, esculin hydrolysis, D-glucose, D-mannose, - to nitrite (NO2 ), reduction of nitrates (NO3) to nitrogen D-mannitol, N-acetyl-D-glucosamine, adipic acid, malic (N2), indole production on tryptophan, glucose fermen- acid, trisodium citrate, and potassium gluconate. Negative - tation, esculin hydrolysis, gelatin hydrolysis, N-ace- for the reduction of nitrates (NO3) to nitrite (NO2 ), reduc- tyl-D-glucosamine, D-maltose, potassium gluconate, tion of nitrates (NO3) to nitrogen (N2), indole production capric acid, adipic acid, and phenylacetic acid. on tryptophan, glucose fermentation, gelatin hydrolysis, In API 32GN system, positive for N-acetyl-glucos- β-galactosidase, L-arabinose, D-maltose, capric acid, and amine, inoitol, L-alanine, D-mannitol, D-glucose, salicin, phenylacetic acid. D-sorbitol, L-arabinose, propionic acid, potassium 2-ke- In API 32GN system, positive for D-ribose, inositol, togluconate, 4-hydroxybenzoic acid, and L-proline. Neg- D-saccharose (sucrose), N-acetyl-glucosamine, suberic ative for L-rhamnose, D-ribose, D-saccharose (sucrose), acid, lactic acid, D-mannitol, D-glucose, salicin, L-arab- D-maltose, itaconic acid, suberic acid, sodium malonate, inose, and 4-hydroxybenzoic acid. Negative for L-rham- sodium acetate, lactic acid, potassium 5-ketogluconate, nose, D-maltose, itaconic acid, sodium malonate, sodi- glycogen, 3-hydroxybenzoic acid, L-serine, D-melibi- um acetate, L-alanine, potassium 5-ketogluconate, gly- ose, D-fucose, capric acid, valeric acid, trisodium citrate, cogen, 3-hydroxybenzoic acid, L-serine, D-melibiose, L-histidine, and 3-hydroxybutyric acid. Strain 16B02D D-fucose, D-sorbitol, propionic acid, capric acid, valeric (=NIBRBAC000500520) was isolated from a soil sam- acid, trisodium citrate, L-histidine, potassium 2-keto- ple. gluconate, 3-hydroxybutyric acid, and L-proline. Strain 16B04G (=NIBRBAC000500521) was isolated from a Description of Oxalicibacterium flavum 16B04G soil sample. Cells are Gram-stain-negative and short rod-shaped. Description of Sphingomonas desiccabilis 16B01D Colonies are pink-colored after 3 days of incubation on R2A at 25°C. In the BIOLOG GEN III, acetoacetic acid, Cells are Gram-stain-negative and short rod-shaped. N-acetyl-D-mannosamine, L-aspartic acid, D-cellobiose, Colonies are yellow-colored after 3 days of incubation on dextrin, D-fructose, D-fructose 6-PO4, L-galactonic acid R2A at 25°C. In the BIOLOG GEN III, dextrin, D-malt- lactone, D-galactose, D-galacturonic acid, gentiobiose, ose, D-trehalose, D-cellobiose, gentiobiose, sucrose, D-gluconic acid, α-D-glucose, D-glucuronic acid, L-glu- D-turanose, D-raffinose, α-D-lactose, D-melibiose, β- tamic acid, α-keto-glutaric acid, L-malic acid, D-malt- methyl-D-glucoside, D-salicin, D-mannose, D-galac- ose, D-mannitol, D-mannose, β-methyl-D-glucoside, tose, D-galacturonic acid, and D-glucuronic acid were 3-methyl glucose, myo-inositol, pectin, propionic acid, utilized as a sole carbon source. But acetoacetic acid, glycyl-L-proline, L-pyroglutamic acid, L-rhamnose, N-acetyl-D-mannosamine, L-alanine, L-arginine, L-as- L-serine, stachyose, sucrose, D-trehalose, D-turanose, partic acid, D-fructose, D-fructose 6-PO4, L-galactonic acetic acid, N-acetyl-neuraminic acid, N-acetyl-D-glu- acid lactone, gelatin, D-gluconic acid, α-D-glucose, cosamine, γ-amino-butryric acid, D-arabitol, D-aspartic glucuronamide, L-glutamic acid, inosine, α-keto-glu- acid, bromo-succinic acid, citric acid, formic acid, D-fu- taric acid, L-malic acid, D-mannitol, 3-methyl glucose, cose, L-fucose, D-glucose-6-PO4, glycerol, L-histidine, myo-inositol, pectin, propionic acid, glycyl-L-proline, α-hydroxybutyric acid, β-hydroxy-D,L-butyric acid, L-pyroglutamic acid, quinic acid, L-rhamnose, L-serine, p-hydroxy-phenylacetic acid, α-keto-butyric acid, L-lac- stachyose, acetic acid, N-acetyl-D-galactosamine, N-acetic acid, D-lactic acid methyl ester, α-D-lactose, D-ma- tyl-neuraminic acid, N-acetyl-D-glucosamine, γ-ami- lic acid, methyl pyruvate, mucic acid, D-saccharic acid, no-butryric acid, D-arabitol, D-aspartic acid, bromo-suc- D-sorbitol, tween 40 are utilized as sole carbon source. cinic acid, citric acid, formic acid, D-fucose, L-fucose, But D-raffinose, D-melibiose, D-salicin, N-acetyl-D-ga- D-glucose-6-PO4, glycerol, L-histidine, α-hydroxybutyr- lactosamine, inosine, D-serine, L-alanine, L-arginine, ic acid, β-hydroxy-D, L-butyric acid, p-hydroxy-pheny- 148 JOURNAL OF SPECIES RESEARCH Vol. 7, No. 2 lacetic acid, α-keto-butyric acid, L-lactic acid, D-lactic acid, methyl pyruvate, D-lactic acid, L-lactic acid, citric acid methyl ester, D-malic acid, methyl pyruvate, mucic acid, α-glutaric acid, D-malic acid, succinic acid, tween acid, D-saccharic acid, D-serine, D-sorbitol, and tween 40, γ-butryric acid, α-butryric acid, β-butryric acid, 40 were not utilized. In sensitivity tests, the tetrazolium acetoacetic acid, propionic acid, acetic acid, and formic redox dye is reduced at pH 6 but not at pH 5; the dye is acid were not utilized. In sensitivity tests, the tetrazoli- reduced in the presence of pH 6, 1% NaCl, 1% sodium um redox dye is reduced at pH 6 but not at pH 5; the dye lactate, rifamycin SV, tetrazolium blue; but not in the is reduced in the presence of 1% NaCl, 4% NaCl, sodi- presence of sodium lactate, 4% NaCl, 8% NaCl, guani- um lactate, D-serine, tetrazolium violet, nalidixic acid, dine HCl, lithium chloride, potassium tellurite, D-serine, lithium chloride, potassium tellurite, aztreonam, and so- sodium butyrate, tetrazolium violet aztreonam, fusidic dium butyrate; but not in the presence of 8% NaCl, fu- acid, lincomycin, minocycline, nalidixic acid, niaproof 4, sidic acid, troleandomycin, rifamycin SV, minocycline, pH 5, sodium bromate, troleandomycin, and vancomy- lincomycin, guanidine HCl, niaproof 4, vancomycin, cin. tetrazolium blue, and sodium bromate. In API 20NE system, positive for of arginine dihydro- In API 20NE system, positive for of reduction of nit lase, urease, esculin hydrolysis, β-galactosidase, D-glu- rates (NO3) to nitrogen (N2), arginine dihydrolase, ure- cose, L-arabinose, D-mannose, N-acetyl-D-glucosamine, ase, esculin hydrolysis, β-galactosidase, D-glucose, L- and D-maltose. Weak positive for D-mannitol, potassi- arabinose, D-mannose, D-mannitol, N-acetyl-D-glu- um gluconate, adipic acid, malic acid, and trisodium ci- cosamine, D-maltose, potassium gluconate, adipic acid, trate. Negative for the reduction of nitrates (NO3) to ni- malic acid, trisodium citrate, and phenylacetic acid. - - trite (NO2 ), reduction of nitrates (NO3) to nitrogen (N2), Negative for reduction of nitrates (NO3) to nitrite (NO2 ), indole production on tryptophan, glucose fermentation, indole production on tryptophan, glucose fermentation, gelatin hydrolysis, capric acid, and phenylacetic acid. gelatin hydrolysis, and capric acid. In API 32GN system, positive for N-acetyl-glucos- In API 32GN system, positive for L-rhamnose, N-ace- amine, D-saccharose (sucrose), D-maltose, suberic acid, tyl-glucosamine, D-ribose, D-saccharose (sucrose), sodium malonate, sodium acetate, lactic acid, L-alanine, D-maltose, suberic acid, sodium acetate, lactic acid, L- potassium 5-ketogluconate, glycogen, 3-hydroxybenzoic alanine, potassium 5-ketogluconate, 3-hydroxybenzo- acid, L-serine, D-glucose, salicin, D-melibiose, D-sorbi- ic acid, D-mannitol, D-glucose, salicin, D-melibiose, tol, L-arabinose, valeric acid, trisodium citrate, L-histi- D-sorbitol, L-arabinose, propionic acid, valeric acid, dine, potassium 2-ketogluconate, 3-hydroxybutyric acid, L-histidine, potassium 2-ketogluconate, 3-hydroxybutyr- 4-hydroxybenzoic acid, and L-proline. Negative for ic acid, 4-hydroxybenzoic acid, and L-proline. Negative L-rhamnose, D-ribose, inositol, itaconic acid, D-man- for inositol, itaconic acid, sodium malonate, glycogen, nitol, D-fucose, propionic acid, and capric acid. Strain L-serine, D-fucose, capric acid, and trisodium citrate. 16B01D (=NIBRBAC000500522) was isolated from a Strain 17U4-2 ( =NIBRBAC000500500) was isolated soil sample. from a soil sample.

Description of Variovorax humicola 17U4-2 Description of Paracoccus acridae 17J28-10 Cells are Gram-stain-positive and short rod-shaped. Cells are Gram-stain-negative and cocci-shaped. Col Colonies are yellow-colored after 3 days of incubation onies are orange-colored after 3 days of incubation on on R2A at 25°C. In the BIOLOG GEN III, D-maltose, R2A at 25°C. In the BIOLOG GEN III, L-alanine, py- D-trehalose, D-cellobiolose, gentiobiose, sucrose, D-tur- roglutamic acid, L-lactic acid, L-malic acid, acetoacetic anose, stachyose, D-raffinose, D-melibiose, β-D-gluco- acid, and acetic acid is utilized as sole carbon source. side, D-salicin, neuraminic acid, α-D-glucose, D-fruc- But D-maltose, D-trehalose, D-cellobiolose, gentiobi- tose, D-mannitol, glycerol, L-histidine, D-gluconic acid, ose, sucrose, D-turanose, stachyose, D-raffinose, D- and L-malic acid were utilized as sole carbon source. melibiose, β-D-glucoside, D-salicin, neuraminic acid, But dextrin, α-D-lactose, N-glucosamine, β-mannosa α-D-glucose, D-fructose, D-mannitol, glycerol, L-histi- min, N-galactosamin, D-mannose, D-galactose, 6-meth- dine, D-gluconic acid, dextrin, α-D-lactose, N-glucos- yl-glucose, D-fucose, L-fucose, L-rhamnose, inosine, amine, β-mannosamin, N-galactosamin, D-mannose, D-sorbitol, D-arabitol, myo-inositol, D-glucose, D-fruc- D-galactose, 6-methyl-glucose, D-fucose, L-fucose, tose, D-aspartic acid, D-serine, gelatin, glycyl-L-proline, L-rhamnose, inosine, D-sorbitol, D-arabitol, myo-ino- L-alanine, L-arginine, L-aspartic acid, L-glutamic acid, sitol, D-glucose, D-fructose, D-aspartic acid, D-serine, pyroglutamic acid, L-serine, pectin, galacturonic acid, gelatin, glycyl-L-proline, L-arginine, L-aspartic acid, L-galactonic lactone, D-glucuronic acid, glucuronamide, L-glutamic acid, L-serine, pectin, galacturonic acid, L-ga- mucic acid, quinic acid, D-saccharic acid, phenylacetic lactonic lactone, D-glucuronic acid, glucuronamide, mu- May 2018 Maeng et al. Report of 11 unrecorded radiation resistant bacteria 149 cic acid, uinic acid, D-saccharic acid, pheylacetic acid, Cao, Y.-R. et al. 2011. Methylobacterium soli sp. nov. a methyl pyruvate, D-lactic acid, citric acid, α-glutaric methanol-utilizing bacterium isolated from the forest acid, D-malic acid, succinic acid, tween40, γ-butryric soil. Antonie Leeuwenhoek 99:629-634. acid, α-butryric acid, β-butryric acid, propionic acid, Doetsch, R. 1981. Determinative methods of light microsco- and formic acid were not utilized. In sensitivity tests, the py. Manual of methods for general bacteriology. pp. 21- tetrazolium redox dye is reduced in the presence of pH 33. 6, 1% NaCl, sodium lactate, and lithium chloride; but Edgar, R.C. 2004. MUSCLE: multiple sequence alignment not in the presence of pH 5, 4% NaCl, 8% NaCl, fusidic with high accuracy and high throughput. Nucleic Acids acid, D-serine, troleandomycin, rifamycin SV, minocy- Res. 32:1792-1797. cline, lincomycin, guanidine HCl, niaproof 4, vancomy- Felsenstein, J. 1985. Confidence limits on phylogenies: an cin, tetrazolium violet, tetrazolium Blue, nalidixic acid, approach using the bootstrap. Evolution 39:783-791. potassium tellurite, aztreonam, sodium butyrate, and so- Gallego, V., M.T. García and A. Ventosa. 2005. Methylobac- dium bromate. terium hispanicum sp. nov. and Methylobacterium aquat- In API 20NE system, positive for of reduction of ni- icum sp. nov., isolated from drinking water. International Journal of Systematic and Evolutionary Microbiology trates (NO3) to nitrogen (N2), arginine dihydrolase, urease, D-glucose, L-arabinose, D-mannose, D-mannitol, 55:281-287. and phenylacetic acid. Negative for reduction of nitrates Garrity G.M., J.A. Bell and T. Lilburn. 2005a. Family IX. - Methylobacteriaceae fam. nov. In: Brenner, D.J., N.R. (NO3) to nitrite (NO2 ), indole production on tryptophan, glucose fermentation, esculin hydrolysis, gelatin hydro- Krieg, J.T. Staley, G.M. Garrity (eds.), Bergey’s manual lysis, β-galactosidase, N-acetyl-D-glucosamine, D-malt- of systematic bacteriology, vol 2, Part C: the alpha- beta-, ose, potassium gluconate, capric acid, adipic acid, malic delta- and Epsilonproteobacteria. Springer, New York, acid, and trisodium citrate. pp. 567-571. In API 32GN system, positive for L-rhamnose, D-ri- Garrity, G.M., J.A. Bell and T. Lilburn. 2005b. Family I. Rhodobacteraceae fam. nov. In: Brenner, D.J., N.R. bose, inositol, lactic acid, L-alanine, potassium 5-keto- Krieg, J.T. Staley and G.M. Garrity (eds.), Bergey’s gluconate, 3-hydroxybenzoic acid, L-serine, D-mannitol, manual of systematic bacteriology, vol 2, 2nd edn, The D-glucose, salicin, D-sorbitol, propionic acid, L-histidine, Proteobacteria, Part C. 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