A Report of 35 Unrecorded Bacterial Species Isolated from Sediment in Korea

Journal362 of Species Research 9(4):362-374, 2020JOURNAL OF SPECIES RESEARCH Vol. 9, No. 4

A report of 35 unrecorded bacterial species isolated from sediment in Korea

Ji-Hye Han1,*, Kiwoon Baek2, Seoni Hwang1, Yoon Jong Nam2 and Mi-Hwa Lee3

1Bacterial Research Team, Microbial Research Department, Nakdonggang National Institute of Biological Resources (NNIBR), Sangju 37242, Republic of Korea 2Bioresources Collection and Research Division, Bioresources Collection and Bioinformation Department, Nakdonggang National Institute of Biological Resources (NNIBR), Sangju 37242, Republic of Korea 3External Relations Division, Exhibition and Education Department, Nakdonggang National Institute of Biological Resources (NNIBR), Sangju 37242, Republic of Korea

*Correspondent: [email protected]

A total of 35 bacterial strains were isolated from various sediment samples. From 16S rRNA gene sequence similarities higher than 98.7% and the formation of a robust phylogenetic clade with the closest species, it was determined that each strain belonged to independent and predefined bacterial species. No previous official reports have described these 35 species in Korea. The unrecorded species were assigned to 6 phyla, 10 classes, 18 orders, 23 families, and 31 genera. At the genus level, the unrecorded species were affiliated with Terriglobus of the phylum Acidobacteria, as well as with Mycobacterium, Rhodococcus, Kineococcus, Phycicoccus, Agromyces, Cryobacterium, Microbacterium, and Arthrobacter; Catellatospora of the class Actinomycetia; Lacibacter of the class Chitinophagia; Algoriphagus and Flectobacillus of the class Cytophagia; Flavobacterium and Maribacter of the class Flavobacteriia; Bacillus, Cohnella, Fontibacillus, Paenibacillus, Lysynibacillus, and Paenisporosarcina of the class Bacilli; Bradyrhizobium, Gemmobacter, Loktanella, and Altererythrobacter of the class Alphaproteobacteria; Acidovorax of the class Betaproteobacteria; Aliiglaciecola, Cellvibrio, Arenimonas, and Lysobacter of class Gammaproteobacteria; and Roseimicrobium of the class Verrucomicrobia. The selected strains were subjected to further taxonomic characterization, including Gram reaction, cellular and colonial morphology, and biochemical properties. This paper provides detailed descriptions of the 35 previously unrecorded bacterial species. Keywords: ‌16S rRNA gene, freshwater, sediment, unrecorded bacteria

Introduction ecosystems. Culture-independent molecular approaches based on Aquatic sediment is one of the largest ecosystems on 16S rRNA have been used for microbial ecology studies Earth. In general, sediments are overlaid by a permanent of sediments and the biomass and taxon richness of mi- water body, such as an ocean, lake, river, or reservoir, and crobes in sediments are known to be much higher than share the property of being continuously wet (Nealson, those of the corresponding water bodies (Wang et al., 1997). Sediment-dwelling microbes are key players in 2012). Although previous studies have been conducted biogeochemical cycling and have potential for the biore- on the basis of traditional cultivation methods, it is now mediation of undesirable compounds from sediments. widely recognized that only 0.001-15% (0.25% in sedi- Microbial communities can be responsible for 76-96% ments) of the total cell counts in environmental samples of the total sediment respiration, and the communities can be cultured (Amann et al., 1995). However, cultiva- are generally known as a dominant driving force in the tion-based study remains important because the ecological transformation of complex organic compounds (Craft et role of bacteria in natural environments and the industrial al., 2002; Zhang et al., 2015). Therefore, investigations application of bacterial resources can be revealed only into the microbial structure and function of sediment is when they are successfully cultivated and characterized. indispensable to obtain a better understanding of aquatic Since 2015, we have isolated a great number of unre- November 2020 Han et al. Unrecorded bacterial species isolated from sediment 363 corded prokaryotic species from the sediments of diverse strains and those of closely related strains was carried out freshwater environments in the Republic of Korea. The using EZ Editor (Jeon et al., 2014). Phylogenetic trees isolated strains were found to belong to six phyla: Acido- were constructed using neighbor-joining (Saitou and Nei, bacteria, Actinobacteria, Bacteroidetes, Firmicutes, Pro- 1987), maximum parsimony (Fitch, 1971), and maximum teobacteria, and Verrucomicrobia. The present report fo- likelihood (Felsenstein, 1981) methods and evolution- cuses on the descriptions of 35 strains that have not been ary distances were calculated using the Jukes and Cantor previously isolated in Korea. one-parameter model (Juke and Cantor, 1969) with the MEGA package ver. 7.0 (Kumar et al., 2016). Bootstrap analysis was carried out using 1,000 resampled datasets Materials and Methods (Felsenstein, 1985), which were also used to test the topology of the phylogenetic tree. Sediment samples were collected from the Nakdong- The colony morphology of the strains was observed gang River, Seomjingang River, Taehwagang River, on agar plates with a magnifying glass after the cells Gyeongpo Lake, Hyang Lake, Chodang Cave, and Moo- grew to stationary phase. Cellular morphology and cell jechineup Wetland (Table 1). Each sediment sample was size were examined by transmission electron microscopy processed separately by spreading onto diverse culture (CM200, Philips or HT7700, Hitachi). Gram staining was agar media: Reasoner’s 2A agar, humic acid vitamin agar, performed using a Gram staining kit (Sigma) and oxi- International Streptomyces Project 4 agar, marine agar, dase activity was determined using 1% (w/v) tetramethyl trypticase soy agar, nutrient agar, and 1/10 diluted agar ρ-phenylenediamine. Biochemical characteristics were of each of these media. After incubation at 15-30°C for tested by using API-20NE galleries (bioMérieux) accord- 7-14 days, bacterial strains were purified by subculturing ing to the manufacturers’ instructions. a single colony on fresh media and pure cultures were preserved at -80°C in 20% (w/v) glycerol suspension. For bacterial identification, PCR amplification with Results and Discussion universal primers 27F and 1492R and 16S rRNA gene sequencing with primers 518F and 800R were performed On the basis of the 16S rRNA gene sequence compar- using standard procedures. The 16S rRNA gene sequenc- ison and phylogenetic analysis, a total of 35 strains were es of the bacterial strains were compared with other assigned to six phyla: one strain to the phylum Acidobacte- bacterial strains using the EZ Biocloud server (Yoon et ria, 10 strains to the phylum Actinobacteria, seven strains al., 2017). Bacterial strains showing 98.7% or higher se- to the phylum Bacteroidetes, six strains to the phylum Fir- quence similarity with validly published species that had micutes, 10 strains to the phylum Proteobacteria, and one not previously been reported in Korea were selected as strain to the phylum Verrucomicrobia. The strain informa- unrecorded bacterial species. For phylogenetic analysis, tion and identification results are summarized in Table 1. 16S rRNA gene sequence alignment between isolated A phylogenetic tree of the 17 bacterial strains assigned

Table 1. GPS information of sampling site for sediment sample collected.

Sampling site Sample code GPS

Lagoon Gyeongpo Lake LS1 N 37º47′49.5″, E 128º53′52.4″ Hyang Lake LS2 N 37º54′30.3″, E 128º48′27.1″

Lime stone cave Chodang Cave CCS N 37º22′47.26″, E 129º10′53.26″

River Nakdonggang River SS N 36º26′4.11″, E 128º15′4.37″ GS N 35º6′4.03″, E 128º56′49.39″ AS N 36º41′38.4″, E 128º48′44.24″ ES N 35º6′42.04″, E 128º56′59.28″

Seomjingang River HS, SJS N 35º3′48.18″, E 127º44′29.21″

Taehwagang River UTS2 N 35º33′7.32″, E 129º19′9.05″ UTS5 N 35º33′5.17″, E 129º18′17.26″

Wetland Moojechineup Wetland UMS N 35º45′91.27″, E 129º13′3.04″ 364 JOURNAL OF SPECIES RESEARCH Vol. 9, No. 4 4 days 3 days 3 days 3 days 3 days 3 days 3 days 3 days 4 days 4 days 3 days 3 days 3 days 3 days 3 days 3 days 3 days 3 days 3 days 4 days 3 days 3 days 3 days 3 days 3 days 3 days 3 days 3 days 3 days 3 days 3 days 3 days ------condition Incubation 15°C, 3 30°C, 2 30°C, 2 30°C, 2 30°C, 2 25°C, 5 days 25°C, 2 20°C, 2 20°C, 3 days 25°C, 2 25°C, 3 30°C, 3 30°C, 2 30°C, 2 25°C, 2 30°C, 2 30°C, 2 37°C, 2 30°C, 2 25°C, 2 10°C, 5 days 20°C, 2 15°C, 3 30°C, 2 30°C, 2 25°C, 2 30°C, 2 30°C, 2 30°C, 2 30°C, 2 25°C, 2 25°C, 2 30°C, 2 30°C, 2 25°C, 2 MA TSA NA R2A TSA R2A R2A R2A R2A R2A R2A TSA R2A R2A R2A R2A R2A R2A TSA R2A MA R2A MA MA NA MA R2A R2A R2A R2A R2A R2A R2A R2A R2A Medium KU341399 KU984671 KY012263 KU984675 KU341402 KY012260 KU341394 KU341392 KY012258 KU984672 KX809752 KX809753 MG780329 MG780328 MK396575 MK396576 MG780339 MK396577 MG780340 MK396581 MK396579 MK396578 MG780336 MG780335 MK396574 MG780337 MG780338 MG780332 MK396584 MK396583 MG780330 MG780331 MK396582 MG780333 MG780334 Accession no.

(%) 98.97 99.06 98.97 99.80 99.04 99.59 99.64 99.93 99.00 99.38 98.78 99.73 99.18 99.93 99.85 99.57 99.80 99.45 98.97 99.22 99.65 99.86 99.49 99.93 99.18 99.73 99.72 99.17 99.51 99.86 99.72 99.29 Sim. 100.00 100.00 100.00 NNIBR No. NNIBR2017301BA33 NNIBR2017301BA32 NNIBR2018141BA3 NNIBR2018141BA4 NNIBR2017301BA43 NNIBR2015296BA10 NNIBR2015296BA9 NNIBR2016301BA21 NNIBR2015296BA38 NNIBR2015296BA15 NNIBR2016301BA27 NNIBR2018141BA5 NNIBR2015296BA4 NNIBR2015296BA2 NNIBR2016301BA24 NNIBR2017301BA44 NNIBR2018141BA9 NNIBR2018141BA7 NNIBR2018141BA6 NNIBR2015296BA35 NNIBR2017301BA28 NNIBR2017301BA40 NNIBR2017301BA39 NNIBR2017301BA29 NNIBR2018141BA2 NNIBR2017301BA41 NNIBR2017301BA42 NNIBR2017301BA36 NNIBR2018141BA12 NNIBR2018141BA11 NNIBR2017301BA34 NNIBR2017301BA35 NNIBR2018141BA10 NNIBR2017301BA37 NNIBR2017301BA38 Strain ID LS1-46 LS1-27 LS1-26 LS2-11 CCS-46 SS1-64 SS1-41 AS4-17 GS1-31 SS2-77 ES2-36 SS5-09 SS1-12 SS1-02 ES1-24 05SS-25 SS12-22 SS7-26 SS7-18 GS1-01 HS1-69 SJS1-74 SJS1-42-1 HS1-70 HS1-68 SJS3-28 SS12-55 UTS6-49 UTS5-20 UTS5-15 UTS2-06 UTS5-52 UTS2-20 UMS-18 UMS-61 Most closely related species Maribacter antarcticus Fontibacillus solani Cellvibrio gandavensis Bacillus niacini Mycolicibacterium rufum Paenibacillus gorillae Gemmobacter aquaticus maotaiensis Arenimonas gellanilyticum Roseimicrobium Algoriphagus ratkowskyi Flavobacterium degerlachei Loktanella atrilutea Acidovorax defluvii Aliiglaciecola litoralis Rhodococcus coprophilus Lacibacter cauensis Flectobacillus major Flavobacterium cauense roseus Terriglobus Cohnella rhizosphaerae hydrothermale Microbacterium kitamiense Microbacterium macmurdoensis Paenisporosarcina taoyuanensis Arenimonas Maribacter arcticus Cryobacterium psychrotolerans enclensis Arthrobacter parviboronicapiens Lysinibacillus Kineococcus endophytica Phycicoccus ginsenosidimutans buctensis Altererythrobacter caeni Lysobacter ramosus Agromyces Catellatospora chokoriensis canariense Bradyrhizobium Family Flavobacteriaceae Paenibacillaceae Cellvibrionaceae Bacillaceae Mycobacteriaceae Paenibacillaceae Rhodobacteraceae Lysobacteraceae Verrucomicrobiaceae Cyclobacteriaceae Flavobacteriaceae Rhodobacteraceae Comamonadaceae Alteromonadaceae Nocardiaceae Chitinophagaceae Cytophagaceae Flavobacteriaceae Acidobacteriaceae Microbacteriaceae Micrococcaceae Planococcaceae Kineosporiaceae Intrasporangiaceae Erythrobacteraceae Lysobacteraceae Microbacteriaceae Micromonosporaceae Nitrobacteraceae Order Flavobacteriales Bacillales Cellvibrionales Bacillales Corynebacteriales Bacillales Rhodobacterales Lysobacterales Verrucomicrobiales Cytophagales Flavobacteriales Rhodobacterales Burkholderiales Alteromonadales Corynebacteriales Chitinophagales Cytophagales Flavobacteriales Acidobacteriales Kineosporiales Micrococcales Sphingomonadales Lysobacterales Micrococcales Micromonosporales Rhizobiales Class Flavobacteria Bacilli Gammaproteobacteria Bacilli Actinomycetia Bacilli Alphaproteobacteria Gammaproteobacteria Verrucomicrobiae Cytophagia Flavobacteria Alphaproteobacteria Betaproteobacteria Gammaproteobacteria Actinomycetia Chitinophagia Cytophagia Flavobacteria Acidobacteriia Alphaproteobacteria Summary of isolated strains from the freshwater sediment and their taxonomic affiliation. Sampling site Hyang Lake Chodang Cave Nakdonggang River Seomjingang River Taehwagang River Moojechineup Wetland Table 2. Table Gyeongpo Lake November 2020 Han et al. Unrecorded bacterial species isolated from sediment 365 to phyla Actinobacteria, Firmicutes, and Verrucomicrobia ter incubation for 2-3 days at 30°C. Positive for esculin is shown in Figure 1. Strains in the phylum Actinobac- hydrolysis and β-galactosidase, but negative for oxidase, teria, Firmicutes, and Verrucomicrobia were assigned to reduction of nitrates, indole production, glucose fermen- the classes Actinomycetia (10 strains), Bacilli (six strains) tation, arginine dihydrolase, urease, and gelatin hydroly- and Verrucomicrobiae (one strain), respectively. At the sis. Does not utilize D-glucose, L-arabinose, D-mannose, genus and family level, 17 strains belonged to 16 genera D-mannitol, N-acetyl-glucosamine, D-maltose, potassium of 11 families: Mycolicibacterium of Mycobacteriaceae, gluconate, capric acid, adipic acid, malic acid, trisodi- Rhodococcus of Nocardiaceae; Kineococcus of Kineospo- um citrate, and phenylacetic acid as sole carbon sources. riaceae; Phycicoccus of Intrasporangiaceae; Agromyces, Strain UMS-18 ( =NNIBR2017301BA37) was isolated Cryobacterium, and Microbacterium of Microbacteriace- from sediment of Moojechineup Wetland, Ulsan, Korea. ae; Arthrobacter of Micrococcaceae; Catellatospora of The GenBank accession number of 16S rRNA gene se- Micromonosporaceae; Bacillus of Bacillaceae; Cohnella, quence is MG780333. Fontibacillus, and Paenibacillus of Paenibacillaceae; Ly- sinibacillus and Paenisporosarcina of Planococcaceae; Phylum Actinobacteria and Roseimicrobium of Verrucomicrobiaceae. The phylogenetic assignments of the strains in the phy- Description of Mycolicibacterium rufum SS1-64 la Acidobacteria, Bacteroidetes, and Proteobacteria are Cells are Gram-stain-positive, non-flagellated, and rod- shown in Figure 2. These 18 strains are belonged to sev- shaped. Colonies grown on Reasoner’s 2A agar (R2A) en classes, including Actinomycetia, Chitinophagia, Cy- are circular, raised, dry with smooth border, and yellow tophagia, Flavobacteriia, Alphaproteobacteria, Betapro- colored after incubation for 5 days at 25°C. Positive for teobacteria, and Gammaproteobacteria. At the generic oxidase, reduction of nitrates, and urease, but negative and family level, 17 strains belonged to 15 genera of 12 for indole production, glucose fermentation, arginine families; Terriglobus of Acidobacteriaceae, Lacibacter of dihydrolase, esculin hydrolysis, and gelatin hydrolysis. Chitinophagaceae, Algoriphagus of Cyclobacteriaceae, D-Mannitol, L-arabinose (weakly), D-maltose (weakly), Flectobacillus of Cytophagaceae, Flavobacterium and potassium gluconate (weakly), and malic acid are utilized Maribacter of Flavobacteriaceae, Bradyrhizobium of Ni- as sole carbon sources, but not D-glucose, D-mannose, trobacteriaceae, Gemmobacter and Loktanella of Rhodo- N-acetyl-glucosamine, capric acid, adipic acid, trisodium bacteraceae, Altererythrobacter of Erythrobacteriaceae, citrate, and phenylacetic acid. Strain SS1-64 ( =NNI- Acidovorax of Comamonadaceae, Aliiglaciecola of Alter- BR2015296BA10) was isolated from sediment of the omonadaceae, Cellvibrio of Cellvibrionaceae, and Areni- Nakdonggang River, Sangju-si, Gyeongsangbuk-do, Ko- monas and Lysobacter of Lysobacteriaceae. rea. The GenBank accession number of 16S rRNA gene These 35 unrecorded species showed diverse cellular sequence is KU341399. morphologies, including rods, short rods, and cocci (Fig. 3). The colony colors were white, cream, ivory, yellow, Description of Rhodococcus coprophilus UTS6-49 orange, or pink. All species grew aerobically and some strains produced spores. The enzymatic and physiological Cells are Gram-stain-positive, non-flagellated, spore properties examined by API 20NE kits were variable de- forming, and rod-shaped. Colonies grown on Reasoner’s pending on species. Detailed cultural, morphological, and 2A agar (R2A) are circular, crateriform, and pinkish-or- biochemical characteristics are also shown in the species ange colored after incubation 2-3 days at 30°C. Positive description section. for reduction of nitrates, and urease in API 20NE, but From the sediments of lagoons, caves, rivers, and wet- negative for oxidase, indole production, glucose fermenta- lands in Korea, 35 species were found and examined tax- tion, arginine dihydrolase, esculin hydrolysis, gelatin hy- onomically. The presence of these 35 species have been drolysis, and β-galactosidase. D-Glucose, and D-mannose reported in overseas, but not previously in Korea. Thus, are utilized as sole carbon sources, but not L-arabinose, the following 35 species are unrecorded species in Korea D-mannitol, N-acetyl-glucosamine, D-maltose, potassium and their phenotypic properties are given. gluconate, capric acid, adipic acid, malic acid, trisodium citrate, and phenylacetic acid. Strain UTS6-49 (=NNI- Phylum Acidobacteria BR2017301BA36) was isolated from sediment of the Taehwagang River, Ulsan, Korea. The GenBank acces- Description of Terriglobus roseus UMS-18 sion number of 16S rRNA gene sequence is MG780332. Cells are Gram-stain-negative, non-flagellated, and Description of Kineococcus endophytica SS1-41 short-rod shaped. Colonies grown on Reasoner’s 2A agar (R2A) are circular, convex, smooth, and pink colored af- Cells are Gram-stain-positive, flagellated, and coc- 366 JOURNAL OF SPECIES RESEARCH Vol. 9, No. 4

Fig. 1. Neighbor-joining phylogenetic tree based on 16S rRNA gene sequences showing the relationship between the strains isolated in this study and their relatives of the phyla Actinobacteria, Firmicutes and Verrucomicrobia. Bootstrap values (expressed as percentages of 1,000 replication) over 50% are shown at node for neighbor-joining, maximum parsimony, maximum likelihood, respectively. Filled circles indicate that the corresponding nodes were also recovered in the trees generated with the maximum likelihood and maximum parsimony algorithms, while open circles indicate that the corresponding nodes were also recovered in the tree generated with one of these algorithms. Bar, 0.05 substitutions per nucleotide position. November 2020 Han et al. Unrecorded bacterial species isolated from sediment 367

Fig. 2. Neighbor-joining phylogenetic tree based on 16S rRNA gene sequences showing the relationship between the strains isolated in this study and their relatives of the phyla Acidobacteria, Bacteroidetes and Proteobacteria. Bootstrap values (expressed as percentages of 1,000 replication) over 50% are shown at node for neighbor-joining, maximum parsimony, maximum likelihood, respectively. Filled circles indicate that the corresponding nodes were also recovered in the trees generated with the maximum likelihood and maximum parsimony algorithms, while open circles indicate that the corresponding nodes were also recovered in the tree generated with one of these algorithms. Bar, 0.05 substitutions per nucleotide position. cus-shaped. Colonies grown on Reasoner’s 2A agar (R2A) after incubation 2-3 days at 25°C. Positive for esculin are circular, smooth, and pale pink to orange colored hydrolysis and β-galactosidase, but negative for reduc- 368 JOURNAL OF SPECIES RESEARCH Vol. 9, No. 4

Fig. 3. Transmission electron micrographs of cells of the strains isolated in this study. Strains: 1, UMS-18; 2, SS1-64; 3, UTS6-49; 4, SS1- 41; 5, AS4-17; 6, UTS5-20; 7, GS1-31; 8, SS2-77; 9, ES2-36; 10, SS5-09; 11, UTS5-15; 12, UTS2-06; 13, HS1-69; 14, UTS5-52; 15, SJS1-74; 16, UTS2-20; 17, LS1-46; 18, SJS1-42-1; 19, CCS-46; 20, LS1-26; 21, LS1-27; 22, SS1-12; 23, SS1-02; 24, ES1-24; 25, UMS- 61; 26, 05SS-25; 27, HS1-70; 28, SS12-22; 29, HS1-68; 30, SJS3-28; 31, LS2-11; 32, SS7-26; 33, SS7-18; 34, SS12-55; 35, GS1-01. November 2020 Han et al. Unrecorded bacterial species isolated from sediment 369 tion of nitrates, indole production, glucose fermentation, tation, arginine dihydrolase, urease, and β-galactosidase. gelatin hydrolysis, and oxidase. D-Glucose, L-arabinose, D-Glucose, L-arabinose, D-mannose, and D-maltose D-mannose, D-maltose, and trisodium citrate are utilized are utilized as sole carbon sources, but not D-mannitol, as sole carbon sources, but not D-mannitol, N-acetyl-glu- N-acetyl-glucosamine, potassium gluconate, capric acid, cosamine, potassium gluconate, capric acid, adipic acid, adipic acid, malic acid, trisodium citrate, and phenylacetic malic acid, and phenylacetic acid. Strain SS1-41 (=NNI- acid. Strain GS1-31 (=NNIBR2015296BA38) was iso- BR2015296BA9) was isolated from sediment of the Na- lated from sediment of the Nakdonggang River, Gumi-si, kdonggang River, Sangju-si, Gyeongsangbuk-do, Korea. Gyeongsangbuk-do, Korea. The GenBank accession The GenBank accession number of 16S rRNA gene se- number of 16S rRNA gene sequence is KU984675. quence is KU984671. Description of Microbacterium hydrothermale SS2-77 Description of Phycicoccus ginsenosidimutans AS4-17 Cells are Gram-stain-positive, non-flagellated, and rod- Cells are Gram-stain-positive, non-flagellated, and coc- shaped. Colonies grown on Reasoner’s 2A agar (R2A) are cus-shaped. Colonies grown on Reasoner’s 2A agar (R2A) circular, opaque, and yellow colored after incubation 2-3 are circular, convex, smooth, and ivory colored after incu- days at 25°C. Positive for reduction of nitrates, esculin bation 2-3 days at 20°C. Positive for reduction of nitrates, hydrolysis, gelatin hydrolysis, and β-galactosidase, but esculin hydrolysis, gelatin hydrolysis, and β-galactosi- negative for oxidase, indole production, glucose fer- dase, but negative for oxidase, indole production, glucose mentation, arginine dihydrolase, and urease. D-Glucose, fermentation, arginine dihydrolase, and urease. D-Glu- L-arabinose, D-mannose, D-mannitol, N-acetyl-glucos- cose, D-mannose, D-mannitol, N-acetyl-glucosamine, amine, D-maltose, and potassium gluconate as sole car- D-maltose, potassium gluconate, and malic acid are uti- bon sources, but not capric acid, adipic acid, malic acid, lized as sole carbon sources, but not L-arabinose, capric trisodium citrate, and phenylacetic acid. Strain SS2-77 acid, adipic acid, trisodium citrate, and phenylacetic acid. (=NNIBR2015296BA15) was isolated from sediment of Strain AS4-17 ( =NNIBR2016301BA21) was isolated the Nakdonggang River, Sangju-si, Gyeongsangbuk-do, from sediment of the Nakdonggang River, Andong-si, Korea. The GenBank accession number of 16S rRNA Gyeongsangbuk-do, Korea. The GenBank accession gene sequence is KU341402. number of 16S rRNA gene sequence is KY012263. Description of Microbacterium kitamiense ES2-36 Description of Agromyces ramosus UTS5-20 Cells are Gram-stain-positive, non-flagellated, and rod- Cells are Gram-stain-positive, non-flagellated, and rod- shaped. Colonies grown on Reasoner’s 2A agar (R2A) are shaped. Colonies grown on Reasoner’s 2A agar (R2A) circular, convex, smooth, and yellow colored after incuba- are circular, convex, smooth, and light-yellow colored tion 3-4 days at 25°C. Positive for esculin hydrolysis, and after incubation 2-3 days at 30°C. Positive for esculin β-galactosidase, but negative for oxidase, reduction of hydrolysis, and β-galactosidase, but negative for oxidase, nitrates, indole production, glucose fermentation, arginine reduction of nitrates, indole production, glucose fermen- dihydrolase, urease, and gelatin hydrolysis. D-Glucose, tation, arginine dihydrolase, urease, and gelatin hydroly- D-mannose, D-mannitol, D-maltose, potassium gluconate sis. L-Arabinose, D-mannitol, and D-maltose are utilized as sole carbon sources, but not L-arabinose, capric acid, as sole carbon sources, but not D-glucose, D-mannose, adipic acid, malic acid, trisodium citrate, and phenylacetic N-acetyl-glucosamine, potassium gluconate, capric acid, acid. Strain ES2-36 (=NNIBR2016301BA27) was iso- adipic acid, malic acid, trisodium citrate, and phenylacetic lated from sediment of the Nakdonggang River, Eulsukdo acid. Strain UTS5-20 (=NNIBR2018141BA12) was iso- Island, Busan, Korea. The GenBank accession number of lated from sediment of the Taehwagang River, Ulsan, Ko- 16S rRNA gene sequence is KY012260. rea. The GenBank accession number of 16S rRNA gene sequence is MK396584. Description of Arthrobacter enclensis SS5-09 Cells are Gram-stain-positive, non-flagellated, rod-, Description of Cryobacterium psychrotolerans GS1-31 and curved rod-shaped. Colonies grown on Trypticase Cells are Gram-stain-positive, non-flagellated, and rod- soy agar (TSA) are circular with entire margin and cream shaped. Colonies grown on Reasoner’s 2A agar (R2A) to light-yellow colored after incubation 3-4 days at are circular, convex with entire edge, and yellow colored 30°C. Positive for oxidase, reduction of nitrates, esculin after incubation 3 days at 20°C. Positive for reduction of hydrolysis, and β-galactosidase, but negative for indole nitrates, esculin hydrolysis, and gelatin hydrolysis, but production, glucose fermentation, arginine dihydrolase, negative for oxidase, indole production, glucose fermen- urease, and gelatin hydrolysis. D-Glucose, L-arabinose, 370 JOURNAL OF SPECIES RESEARCH Vol. 9, No. 4

D-mannose, and D-mannitol are utilized as sole carbon fermentation, arginine dihydrolase, and urease. Does not sources, but not N-acetyl-glucosamine, D-maltose, po- utilize D-glucose, L-arabinose, D-mannose, D-mannitol, tassium gluconate, capric acid, adipic acid, malic acid, N-acetyl-glucosamine, D-maltose, potassium gluconate, trisodium citrate, and phenylacetic acid. Strain SS5-09 capric acid, adipid acid, malic acid, trisodium citrate, and (=NNIBR2018141BA5) was isolated from sediment of phenylacetic acid as sole carbon sources. Strain HS1- the Nakdonggang River, Sangju-si, Gyeongsangbuk-do, 69 ( =NNIBR2017301BA28) was isolated from sedi- Korea. The GenBank accession number of 16S rRNA ment of the Seomjingang River, Hadong-gun, Gyeong- gene sequence is MK396577. sangnam-do, Korea. The GenBank accession number of 16S rRNA gene sequence is KX809752. Description of Catellatospora chokoriensis UTS5-15 Description of Flectobacillus major UTS5-52 Cells are Gram-stain-positive, non-flagellated, spore- forming, and rod-shaped. Colonies grown on Reasoner’s Cells are Gram-stain-negative, non-flagellated, and rod- 2A agar (R2A) are circular, convex, smooth, and light-yel- shaped. Colonies grown on Reasoner’s 2A agar (R2A) are low colored after incubation 2-3 days at 30°C. Positive irregular, convex, smooth, mucoid, and light-pink colored for esculin hydrolysis, gelatin hydrolysis, and β-galac- after incubation 2-3 days at 25°C. Positive for oxidase, tosidase, but negative for oxidase, reduction of nitrates, reduction of nitrates, esculin hydrolysis, and β-galactosi- indole production, glucose fermentation, arginine dihy- dase, but negative for indole production, glucose fermen- drolase, and urease. D-Glucose, L-arabinose, D-mannose, tation, arginine dihydrolase, urease, and gelatin hydroly- and D-maltose are utilized as sole carbon sources, but not sis. D-Glucose, L-arabinose, D-mannose, N-acetyl-glu- D-mannitol, N-acetyl-glucosamine, potassium gluconate, cosamine, D-maltose, and potassium gluconate as sole capric acid, adipic acid, malic acid, trisodium citrate, carbon sources, but not D-mannitol, capric acid, adipic and phenylacetic acid. Stain UTS5-15 ( =NNIBR2018 acid, malic acid, trisodium citrate, and phenylacetic acid. 141BA11) was isolated from sediment of the Taehwagang Strain UTS5-52 (=NNIBR2017301BA35) was isolated River, Ulsan, Korea. The GenBank accession number of from sediment of the Taehwagang River, Ulsan, Korea. 16S rRNA gene sequence is MK396583. The GenBank accession number of 16S rRNA gene sequence is MG780331. Phylum Bacteroidetes Description of Flavobacterium degerlachei SJS1-74 Description of Lacibacter cauensis UTS2-06 Cells are Gram-stain-negative, non-flagellated, and rod- Cells are Gram-stain-negative, non-flagellated, and rod- shaped. Colonies grown on Reasoner’s 2A agar (R2A) are shaped. Colonies grown on Reasoner’s 2A agar (R2A) are circular, convex, smooth, and yellow colored after incu- circular, convex, smooth, and orange colored after incubation 2-3 days at 20°C. Positive for esculin hydrolysis bation 2-3 days at 25°C. Positive for oxidase, esculin hy- and gelatin hydrolysis, but negative for oxidase, reduc- drolysis, gelatin hydrolysis, and β-galactosidase, but neg- tion of nitrates, indole production, glucose fermentation, ative for reduction of nitrates, indole production, glucose arginine dihydrolase, and β-galactosidase. D-Glucose, fermentation, arginine dihydrolase, and urease. D-Glu- D-mannose, and D-maltose are utilized as sole carbon cose, D-mannose, N-acetyl-glucosamine, and D-maltose sources, but not L-arabinose, D-mannitol, N-acetyl-glu- are utilized as sole carbon sources, but not L-arabinose, cosamine, potassium gluconate, capric acid, adipic acid, D-mannitol, potassium gluconate, capric acid, adipic acid, malic acid, trisodium citrate, and phenylacetic acid. Strain malic acid, trisodium citrate, and phenylacetic acid. Strain SJS1-74 ( =NNIBR2017301BA40) was isolated from UTS2-06 ( =NNIBR2017301BA34) was isolated from sediment of the Seomjingang River, Hadong-gun, Gyeo- sediment the Taehwagang River, Ulsan, Korea. The Gen- ngsangnam-do, Korea. The GenBank accession number Bank accession number of 16S rRNA gene sequence is of 16S rRNA gene sequence is MG780336. MG780330. Description of Flavobacterium cauense UTS2-20 Description of Algoriphagus ratkowskyi HS1-69 Cells are Gram-stain-negative, flagellated, and rod- Cells are Gram-stain-negative, non-flagellated, and shaped. Colonies grown on Reasoner’s 2A agar (R2A) are short rod-shaped. Colonies grown on Marine agar (MA) circular, convex, smooth, and yellow colored after incuba- are circular, convex, smooth, and orange colored after in- tion 2-3 days at 30°C. Positive for arginine dihydrolase, cubation 5 days at 10°C. Positive for oxidase, esculin hy- esculin hydrolysis (weakly), gelatin hydrolysis, and β-ga- drolysis, gelatin hydrolysis, and β-galactosidase, but neg- lactosidase (weakly), but negative for oxidase, reduction ative for reduction of nitrates, indole production, glucose of nitrates, indole production, glucose fermentation, and November 2020 Han et al. Unrecorded bacterial species isolated from sediment 371 urease. Does not utilize D-glucose, L-arabinose, D-man- glucose fermentation, arginine dihydrolase, and gelatin nose, D-mannitol, N-acetyl-glucosamine, D-maltose, hydrolysis. D-Glucose, N-acetyl-glucosamine (weak- potassium gluconate, capric acid, adipic acid, malic acid, ly), D-maltose, and potassium gluconate are utilized as trisodium citrate, and phenylacetic acid as sole carbon sole carbon sources, but not L-arabinose, D-mannose, sources. Strain UTS2-20 (=NNIBR2018141BA10) was D-mannitol, capric acid, adipic acid, malic acid, trisodi- isolated from sediment the Taehwagang River, Ulsan, Ko- um citrate, and phenylacetic acid. Strain CCS-46 (=NNI- rea. The GenBank accession number of 16S rRNA gene BR2017301BA43) was isolated from sediment of the sequence is MK396582. Chodang Cave, Samcheok-si, Gangwon-do, Korea. The GenBank accession number of 16S rRNA gene sequence Description of Maribacter antarcticus LS1-46 is MG780339.

Cells are Gram-stain-negative, non-flagellated, and rod- Description of Cohnella rhizospherae LS1-26 shaped. Colonies grown on Marine agar (MA) are circular, convex, smooth, and yellow colored after incubation Cells are Gram-stain-positive, non-flagellated, and rod- 3-4 days at 15°C. Positive for glucose fermentation, shaped. Colonies grown on Nutrient agar (NA) are circu- esculin hydrolysis, and β-galactosidase, but negative for lar, convex, and ivory colored after incubation 2-3 days at oxidase, reduction of nitrates, indole production, arginine 30°C. Positive for oxidase, reduction of nitrates, esculine dihydrolase, urease, and gelatin hydrolysis. Does not uti- hydrolysis, and β-galactosidase, but negative for indole lize D-glucose, L-arabinose, D-mannose, D-mannitol, production, glucose fermentation, arginine dihydrolase, N-acetyl-glucosamine, D-maltose, potassium gluconate, urease, and gelatin hydrolysis. D-Glucose, L-arabinose, capric acid, adipic acid, malic acid, trisodium citrate, and D-mannose, and D-mannitol are utilized as sole carbon phenylacetic acid as sole carbon sources. Strain LS1-46 sources, but not N-acetyl-glucosamine, D-maltose, po- (=NNIBR2017301BA33) was isolated from sediment of tassium gluconate, capric acid, adipic acid, malic acid, the Gyeongpo Lake, Gangneung-si, Gangwon-do, Korea. trisodium citrate, and phenylacetic acid. Strain LS1-26 The GenBank accession number of 16S rRNA gene se- (=NNIBR2018141BA3) was isolated from sediment of quence is MG780329. the Gyeongpo Lake, Gangneung-si, Gangwon-do, Korea. The GenBank accession number of 16S rRNA gene se- Description of Maribacter arcticus SJS1-42-1 quence is MK396575. Cells are Gram-stain-negative, non-flagellated, and rod- Descriptioni of Fontibacillus solani LS1-27 shaped. Colonies grown on Marine agar (MA) are circular, convex, smooth, and yellow colored after incubation Cells are Gram-stain-positive, non-flagellated, and rod- 3-4 days at 15°C. Positive for glucose fermentation, argi- shaped. Colonies grown on trypticase soy agar (TSA) are nine dihydrolase, esculin hydrolysis, and β-galactosidase, circular, convex, smooth, and ivory colored after incuba- but negative for oxidase, reduction of nitrates, indole tion 2-3 days at 30°C. Positive for glucose fermentation, production, urease, and gelatin hydrolysis. D-Glucose, esculin hydrolysis, and β-galactosidase, but negative for D-mannose, N-acetyl-glucosamine, D-maltose, potassium oxidase, reduction of nitrates, indole production, arginine gluconate, adipic acid (weakly), and malic acid are uti- dihydrolase, urease, and gelatin hydrolysis. L-Arabinose lized as sole carbon sources, but not L-arabinose, D-man- and D-maltose are utilized as sole carbon sources, but nitol, capric acid, trisodium citrate, and phenylacetic acid. not D-glucose, D-mannose, D-mannitol, N-acetyl-glu- Strain SJS1-42-1 (=NNIBR2017301BA39) was isolated cosamine, potassium gluconate, capric acid, adipic acid, from sediment of the Seomjingang River, Hadong-gun, malic acid, trisodium citrate, and phenylacetic acid. Strain Gyeongsangnam-do, Korea. The GenBank accession LS1-27 ( =NNIBR2017301BA32) was isolated from number of 16S rRNA gene sequence is MG780335. sediment of the Gyeongpo Lake, Gangneung-si, Gang- won-do, Korea. The GenBank accession number of 16S Phylum Firmicutes rRNA gene sequence is MG780328.

Description Bacillus niacini CCS-46 Description of Paenibacillus gorillae SS1-12 Cells are Gram-stain-positive, non-flagellated, and rod- Cells are Gram-stain-positive, flagellated, and rod- shaped. Colonies grown on Trypticase soy agar (TSA) shaped. Colonies grown on Reasoner’s 2A agar (R2A) are circular, convex, smooth, and greyish-ivory colored are irregular and white-cream colored after incubation after incubation 2-3 days at 30°C. Positive for reduction 2-3 days at 30°C. Positive for urease, esculin hydrolysis, of nitrates, urease, esculin hydrolysis (weakly), and β-ga- and β-galactosidase, but negative for oxidase, reduction lactosidase, but negative for oxidase, indole production, of nitrates, indole production, glucose fermentation, ar- 372 JOURNAL OF SPECIES RESEARCH Vol. 9, No. 4 ginine dihydrolase, and gelatin hydrolysis. D-Glucose, nitrates, arginine dihydrolase, urease, esculin hydrolysis, L-arabinose, D-mannose, D-mannitol, N-acetyl-glucos- and β-galactosidase, but negative for indole production, amine, D-maltose, and potassium gluconate are utilized glucose fermentation, and gelatin hydrolysis. D-Glucose, as sole carbon sources, but not capric acid, adipic acid, L-arabinose, D-mannose, D-mannitol, potassium gluco- malic acid, trisodium citrate, and phenylacetic acid. Strain nate, adipic acid, malic acid, and trisodium citrate, but not SS1-12 (=NNIBR2015296BA4) was isolated from sedi- N-acetyl-glucosamine, D-maltose, capric acid, and phe- ment of the Nakdonggang River, Sangju-si, Gyeongsang- nylacetic acid. Strain UMS-61 (=NNIBR2017301BA38) buk-do, Korea. The GenBank accession number of 16S was isolated from sediment of the Moojechineup Wet- rRNA gene sequence is KU341394. land, Ulsan, Korea. The GenBank accession number of 16S rRNA gene sequence is MG780334. Description of Lysinibacillus parviboronicapiens SS1-02 Description of Gemmobacter aquaticus 05SS-25 Cells are Gram-stain-positive, non-flagellated, and rod- shaped. Colonies grown on Reasoner’s 2A agar (R2A) are Cells are Gram-stain-negative, non-flagellated, and rod- circular with entire edges, opaque, and cream colored af- shaped. Colonies grown on Reasoner’s 2A agar (R2A) are ter incubation 2-3 days at 30°C. Positive for arginine di- circular, convex, smooth, and cream colored after incuba- hydrolase, urease, and gelatin hydrolysis, but negative for tion 2-3 days at 30°C. Positive for reduction of nitrates oxidase, reduction of nitrates, indole production, glucose to nitrogen, arginine dihydrolase, urease, esculin hydroly- fermentation, esculin hydrolysis, and β-galactosidase. sis, and β-galactosidase, but negative for oxidase, indole N-Acetyl-glucosamine, malic acid, and phenylacetic acid production, glucose fermentation, and gelatin hydrolysis. are utilized as sole carbon sources, but not D-glucose, N-Acetyl-glucosamine, D-maltose, potassium gluconate, A-arabinose, D-mannose, D-mannitol, D-maltose, potas- adipic acid, malic acid, trisodium citrate, and phenylacetic sium gluconate, capric acid, adipic acid, and trisodium acid are utilized as sole carbon sources, but not D-glu- citrate. Strain SS1-02 (=NNIBR2015296BA2) was iso- cose, L-arabinose, D-mannose, D-mannitol, and capric lated from sediment of the Nakdonggang River, Sangju- acid. Strain 05SS-25 (=NNIBR2017301BA44) was iso- si, Gyeongsangbuk-do, Korea. The GenBank accession lated from sediment of the Nakdonggang River, Sangju- number of 16S rRNA gene sequence is KU341392. si, Gyeongsangbuk-do, Korea. The GenBank accession number of 16S rRNA gene sequence is MG780340. Description of Paenisporosarcina macmurdoensis ES1-24 Description of Loktanella atrilutea HS1-70 Cells are Gram-stain-positive, non-flagellated, and rod- Cells are Gram-stain-negative, non-flagellated, and rod- shaped. Colonies grown on Reasoner’s 2A agar (R2A) are shaped. Colonies grown on marine agar (MA) are circular, circular, flat, irregular, opaque, and cream colored after convex, smooth, and ivory colored after incubation 2-3 incubation 2-3 days at 25°C. Positive for oxidase, esculin days at 30°C. Positive for oxidase, glucose fermentation, hydrolysis, and gelatin hydrolysis, but negative for reduc- urease, esculin hydrolysis, and β-galactosidase, but nega- tion of nitrates, indole production, glucose fermentation, tive for reduction of nitrates, indole production, arginine arginine dihydrolase, urease, and β-galactosidase. D-Glu- dihydrolase, and gelatin hydrolysis. D-Glucose, L-arab- cose, and potassium gluconate are utilized as sole carbon inose, D-mannose, D-mannitol (weakly), and D-maltose sources, but not L-arabinose, D-mannose, D-mannitol, are utilized as sole carbon sources, but not N-acetyl-glu- N-acetyl-glucosamine, D-maltose, capric acid, adipic cosamine, potassium gluconate, capric acid, adipic acid, acid, malic acid, trisodium citrate, and phenylacetic acid. malic acid, trisodium citrate, and phenylacetic acid. Strain Strain ES1-24 ( =NNIBR2016301BA24) was isolated HS1-70 ( =NNIBR2017301BA29) was isolated from from sediment of the Nakdonggang River, Eulsukdo Is- sediment of the Seomjingang River, Hadong-gun, Gyeo- land, Busan, Korea. The GenBank accession number of ngsangnam-do, Korea. The GenBank accession number 16S rRNA gene sequence is KY012258. of 16S rRNA gene sequence is KX809753.

Phylum Proteobacteria Description of Altererythrobacter buctensis SS12-22 Cells are Gram-stain-negative, non-flagellated, and rod- Description of Bradyrhizobium canariense UMS-61 shaped. Colonies grown on Reasoner’s 2A agar (R2A) Cells are Gram-stain-negative, non-flagellated, and rod- are circular, convex, smooth, mucoid, and yellow colored shaped. Colonies grown on Reasoner’s 2A agar (R2A) are after incubation 2-3 days at 30°C. Positive for oxidase, circular, convex, smooth, and white colored after incuba- esculin hydrolysis, and β-galactosidase, but negative for tion 2-3 days at 25°C. Positive for oxidase, reduction of reduction of nitrates, indole production, glucose fermen- November 2020 Han et al. Unrecorded bacterial species isolated from sediment 373 tation, arginine dihydrolase, urease, and gelatin hydroly- oxidase, reduction of nitrates, indole production, glucose sis. D-Glucose, L-arabinose, D-mannose, and D-maltose fermentation, arginine dihydrolase, urease, and gelatin hy- are utilized as sole carbon sources, but not D-mannitol, drolysis. Does not utilize D-glucose, L-arabinose, D-man- N-acetyl-glucosamine, potassium gluconate, capric acid, nose, D-mannitol, N-acetyl-glucosamine, D-maltose, adipic acid, malic acid, trisodium citrate, and phaeny- potassium gluconate, capric acid, adipic acid, malic acid, lacetic acid. Strain SS12-22 ( =NNIBR2018141BA9) tirsocium citrate, and phenylacetic acid as sole carbon was isolated from sediment of the Nakdonggang Riv- sources. Strain LS2-11 (=NNIBR2018141BA4) was iso- er, Sangju-si, Gyeongsangbuk-do, Korea. The Gen- lated from sediment of the Hyang-Lake, Gangneung-si, Bank accession number of 16S rRNA gene sequence is Gangwon-do, Korea. The GenBank accession number of MK396581. 16S rRNA gene sequence is MK396576.

Description of Acidovorax defluvii HS1-68 Description of Arenimonas maotaiensis SS7-26 Cells are Gram-stain-negative, flagellated, and rod- Cells are Gram-stain-negative, non-flagellated, and rod- shaped. Colonies grown on nutrient agar (NA) are circular shaped. Colonies grown on Reasoner’s 2A agar (R2A) are with entire margin, convex, smooth and cream-white col- circular, convex, translucent, smooth, and white colored ored after incubation 2-3 days at 30°C. Positive for oxi- after incubation 2-3 days at 37°C. Positive for oxidase, dase, reduction of nitrates to nitrogen, urease, and esculin esculin hydrolysis, and gelatin hydrolysis, but negative hydrolysis (weakly), but negative for indole production, for reduction of nitrates, indole production, glucose fer- glucose fermentation, arginine dihydrolase, gelatin hy- mentation, arginine dihyrolase, urease, and β-galactosi- drolysis, and β-galactosidase. D-Mannitol, N-acetyl-glu- dase. Does not utilize D-glucose, L-arabinose, D-man- cosamine, potassium gluconate, capric acid, adipic acid, nose, D-mannitol, N-acetyl-glucosamine, D-maltose, and malic acid are utilized as sole carbon sources, but not potassium gluconate, capric acid, adipic acid, malic acid, D-glucose, L-arabinose, D-mannose, D-maltose, trisodi- tirsocium citrate, and phenylacetic acid as sole carbon um citrate, and phenylacetic acid. Strain HS1-68 (=NNI- sources. Strain SS7-26 (=NNIBR2018141BA7) was iso- BR2018141BA2) was isolated from sediment of the lated from sediment of the Nakdonggang River, Sangju- Seomjingang River, Hadong-gun, Gyeongsangnam-do, si, Gyeongsangbuk-do, Korea. The GenBank accession Korea. The GenBank accession number of 16S rRNA number of 16S rRNA gene sequence is MK396579. gene sequence is MK396574. Description of Arenimonas taoyuanensis SS7-18 Description of Aliiglaciecola litoralis SJS3-28 Cells are Gram-stain-negative, non-flagellated, and rod- Cells are Gram-stain-negative, flagellated, and rod- shaped. Colonies grown on trypticase soy agar (TSA) are shaped. Colonies grown on marine agar (MA) are circu- circular with margins, convex, opaque, and ivory colored lar, convex, smooth and ivory colored after incubation after incubation 2-3 days at 30°C. Positive for oxidase, 2-3 days at 25°C. Positive for esculin hydrolysis, gelatin and gelatin hydrolysis, but negative for reduction of ni- hydrolysis, and β-galactosidase, but negative for oxidase, trates, indole production, glucose fermentation, arginine reduction of nitrates, indole production, glucose fermen- dihydrolase, urease, esculin hydrolysis, and β-galactosi- tation, and arginine dihydrolase. D-Glucose, L-arabinose, dase. Does not utilize D-glucose, L-arabinose, D-man- D-mannose, D-mannitol, D-maltose, potassium gluco- nose, D-mannitol, N-acetyl-glucosamine, D-maltose, nate, adipic acid, malic acid, and trisodium citrate are potassium gluconate, capric acid, adipic acid, malic acid, utilized as sole carbon sources, but not N-acetyl-glucos- tirsocium citrate, and phenylacetic acid as sole carbon amine, capric acid, and phenylacetic acid. Strain SJS3- sources. Strain SS7-18 (=NNIBR2018141BA6) was iso- 28 ( =NNIBR2017301BA41) was isolated from sedi- lated from sediment of the Nakdonggang River, Sangju- ment of the Seomjingang River, Hadong-gun, Gyeong- si, Gyeongsangbuk-do, Korea. The GenBank accession sangnam-do, Korea. The GenBank accession number of number of 16S rRNA gene sequence is MK396578. 16S rRNA gene sequence is MG780337. Description of Lysobacter caeni SS12-55 Description of Cellvibrio gandavensis LS2-11 Cells are Gram-stain-negative, non-flagellated, and rod- Cells are Gram-stain-negative, non-flagellated, and rod- shaped. Colonies grown on Reasoner’s 2A agar (R2A) are shaped. Colonies grown on Reasoner’s 2A agar (R2A) circular, convex, smooth, and ivory to brown colored after are irregular, convex, opaque, shiny, mucoid, and ivory incubation 2-3 days at 30°C. Positive for oxidase, reduc- colored after incubation 2-3 days at 30°C. Positive for tion of nitrates to nitrogen, esculin hydrolysis, and gelatin esculin hydrolysis and β-galactosidase, but negative for hydrolysis, but negative for indole production, glucose 374 JOURNAL OF SPECIES RESEARCH Vol. 9, No. 4 fermentation, arginine dihydrolase, urease, and β-galac- river. Freshwater Biology 47:251-261. tosidase. D-Glucose and D-maltose are utilized as sole Felsenstein, J. 1981. Evolutionary trees from DNA sequences: carbon sources, but not L-arabinose, D-mannose, D-man- a maximum likelihood approach. Journal of Molecular nitol, N-acetyl-glucosamine, potassium gluconate, capric Evolution 17:368-376. acid, adipic acid, malic acid, trisodium citrate, and phe- Felsenstein, J. 1985. Confidence limits on phylogenies: An nylacetic acid. Strain SS12-55 (=NNIBR2017301BA42) approach using the bootstrap. Evolution 39:783-791. was isolated from sediment of the Nakdonggang Riv- Fitch, W.M. 1971. Toward defining the course of evolution: er, Sangju-si, Gyeongsangbuk-do, Korea. The Gen- minimum change for a specific tree topology. Systematic Bank accession number of 16S rRNA gene sequence is Zoology 20:406-416. MG780338. Jeon, Y.-S., K. Lee, S.-C. Park, B.-S. Kim, Y.-J. Cho, S.-M. Ha and J. Chun. 2014. EzEditor: a versatile sequence Phylum Verrucomicrobia alignment editor for both rRNA- and protein-coding genes. International Joural of Systematic and Evolutionary Description of Roseimicrobium gellanilyticum GS1-01 Microbiology 64:689-691. Jukes, T.H. and C.R. Cantor. 1969. Evolution of protein mole- Cells are Gram-stain-negative, non-flagellated, and rod- cules In: H.N. Munro (ed.), Mammaliam protein metabo- shaped. Colonies grown on Reasoner’s 2A agar (R2A) are lism Vol. 3. Academic Press, New York: 21-213. circular, convex, smooth, and pink to brown colored after Kumar, S., G. Stecher and K. Tamura. 2016. MEGA7: Molec- incubation 2-3 days at 25°C. Positive for oxidase, urease, ular Evolutionary Genetics Analysis Version 7.0 for Big- esculin hydrolysis, and β-galactosidase, but negative for ger Datasets. Molecular Biology and Evolution 33:1870- reduction of nitrates, indole production, glucose fermenta- 1874. tion, arginine dihydrolase, and gelatin hydrolysis. D-Glu- Nealson, K.H. 1997. Sediment bacteria : who’s there, what are cose and D-mannose are utilized as sole carbon sources, they doing, and what’s new? Annual Review of Earth and but not L-arabinose, D-mannitol, N-acetyl-glucosamine, Planetary Sciences 25:403-434. D-maltose, potassium gluconate, capric acid, adipic acid, Saitou, N. and M. Nei. 1987. The neighbor-joining method: a malic acid, trisodium citrate, and phenylacetic acid. Strain new method for reconstructing phylogenetic trees. Molec- GS1-01 (=NNIBR2015296BA35) was isolated from sed- ular Biology and Evolution 4:406-425. iment of the Nakdonggang River, Gumi-si, Gyeongsang- Wang, Y., H.-F. Sheng, Y. He, J.-Y. Wu, Y.-X. Jiang, N.F.-Y. buk-do, Korea. The GenBank accession number of 16S Tam and H.-W. Zhou. 2012. Comparison of the levels of rRNA gene sequence is KU984672. bacterial diversity in freshwater, intertidal wetland, and marine sediments by using millions of illumina tags. Ap- plied and Environmental Microbiology 78:8264-8271. Acknowledgements Yoon, S.-H., S.-M. Ha, S. Kwon, J. Lim, Y. Kim, H. Seo and J. Chun. 2017. Introducing EzBioCloud: A taxonomically This study was supported by a grant from the Nakdong- united database of 16S rRNA and whole genome assem- gang National Institute of Biological Resources (NNIBR), blies. International Journal of Systematic and Evolution- funded by the Ministry of Environment (MOE) of the Re- ary Microbiology 67:1613-1617. public of Korea (NNIBR202001106). Zhang, X., Q. Gu, X.-E. Long, Z.-L. Li, D.-X. Liu, D.-H. Ye, C.-Q. He, X.-Y. Liu, K. Väänänen and X.-P. Chen. 2016. Anthropogenic activities drive the microbial community and its function in urban river sediment. Journal of Soils References and Sediments 16:716-725. Amann, R.I., W. Ludwig and K.-H. Schleifer. 1995. Phyloge- netic identification and in situ detection of individual microbial cells without cultivation. Microbiological Reviews 59:143-169. Submitted: August 4, 2020 Craft, J.A., J.A. Stanford and M. Pusch. 2002. Microbial res- Revised: October 19, 2020 piration within a floodplain aquifer of a large gravel-bed Accepted: October 20, 2020