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ISSN : 1226-9999(Print)
ISSN : 2287-7851(Online)
Korean J. Environ. Biol. Vol.41 No.3 pp.308-324
DOI : https://doi.org/10.11626/KJEB.2023.41.3.308

A report of 44 unrecorded bacterial species isolated from Nakdong River in Korea

Ju-Hyung Jeon, Sanghwa Park, Ja Young Cho, Soo-Yeong Lee, Seoni Hwang, Jun Sung Kim, Eui-Jin Kim, Ji Young Jung*
Microbial Research Department, Nakdonggang National Institute of Biological Resources, Sangju 37242, Republic of Korea
* Corresponding author Ji Young Jung Tel. 054-530-0866 E-mail. jyjung@nnibr.re.kr

Contribution to Environmental Biology


▪ A total of 44 unrecorded bacterial species in Korea were newly reported.


▪ This study expands our knowledge of the bacterial diversity in the freshwater habitat of Korea.


31/07/2023 15/09/2023 20/09/2023

Abstract


This study investigated unrecorded freshwater bacterial species in Korea. Water and sediment samples were collected from the Nakdong River basin from 2020-2022. Bacterial isolates obtained through the conventional culture method with commercial media were subjected to 16S rRNA gene sequencing to identify unrecorded bacterial species. Results of 16S rRNA gene sequencing of the bacterial isolates revealed that a total of 44 bacterial isolates shared 16S rRNA gene sequence similarities of more than 98.65%, with validly published bacterial species not reported in Korea yet. These isolates were phylogenetically assigned to 4 phyla, 7 classes, 21 orders, 33 families, and 42 genera. A total of 2, 6, 12, and 24 species belonged to phyla Bacillota, Bacteroidota, Actinomycetota, and Pseudomonadota, respectively. Here, we provide details of these 44 unrecorded bacterial species, including Gram staining, colony and cellular morphologies, biochemical properties, and phylogenetic position.


freshwater , Nakdong River basin , indigenous bacterial species , unrecorded bacterial species in Korea , 16S rRNA gene sequence

초록

    1. INTRODUCTION

    Freshwater habitats are a tremendous source of organic matter. They play a vital role in global biogeochemical cycles. Microorganisms contribute to the biogeochemical cycle by degrading refractory organic substances and recycling nutrients, carbon, nitrogen, sulfur, phosphorus, minerals, and toxic elements in freshwater ecosystems (Arora-Williams et al. 2018;Linz et al. 2018;Zak et al. 2021). Numerous studies have been conducted to understand microbial diversity and ecological functions of microbes in freshwater environment (Grossart et al. 2019;Tran et al. 2021;Kim et al. 2022;Zhao et al. 2023). In particular, microbes with key roles in biogeochemical cycles have been isolated with their functional traits characterized (Ghai et al. 2014;Henson et al. 2018). In addition, freshwater microorganisms as sources of hydrolytic enzymes, functional secondary metabolites, pollutant degrading enzymes, bioflocculants, and nutritional foods, are being studied for eco-friendly applications in various industrial sectors (Mabinya et al. 2012;Suhadolnik et al. 2017;Chantarasiri 2021;Boukid and Castellari 2022;Graffius et al. 2023).

    As part of the research program conducted by Nakdonggang National Institute of Biological Resources (NNIBR), numerous bacterial strains were isolated from the freshwater samples to broaden our knowledge of bacterial diversity and their functional characteristics and novel and/or unrecorded bacterial species inhabited in Korea were discovered. Here, we report a total of 44 bacterial species that have not yet been recorded in Korea. They were isolated from environmental samples collected from Nakdong River basin. These bacterial species belong to phyla Actinomycetota, Bacillota, Bacteroidota, and Pseudomonadota based on phylogenetic analysis.

    2. MATERIALS AND METHODS

    Samples of freshwater and sediment in stream were collected from a total of 16 sites in Nakdong River basin of Korea. These samples were serially diluted in sterile phosphate-buffered saline (PBS, pH 7.4), and then inoculated onto Reasonerʹs 2A agar (R2A; Difco, USA), 1/10 R2A, nutrient agar (NA; Difco, USA), 1/10 NA, tryptic soy agar (TSA; Difco, USA), 1/10 TSA, marine agar (MA; Difco, USA), 1/10 MA, 1/10 MA supplemented 0.03% (w/v) sodium phosphate, and inorganic salt-starch agar (ISP no. 4; MB cells, Republic of Korea) plate. These plates were then, aerobically incubated at 20°C for two weeks. Single colonies grown on each agar plate were transferred by streaking cells onto a new agar medium to obtain pure bacterial cultures.

    16S rRNA genes of the pure-cultured bacterial cells were individually PCR-amplified and sequenced as previously described (Jung et al. 2023). Obtained 16S rRNA gene sequences were then compared with sequences of all reported bacterial type stains in EzBio- Cloud DB (Yoon et al. 2017) to calculate pairwise sequence similarities. Bacterial strains sharing 16S rRNA gene sequence similarity more than 98.65% with type strains of any bacterial species with valid name, which have not been reported in Korea, were identified as bacterial species unreported in Korea. These identified unreported bacterial species were subjected to phylogenetic and phenotypic analyses as described below.

    For phylogenetic analyses, 16S rRNA gene sequences of unreported bacterial species and closely related type strains were multiple-aligned using CLUSTAL W (Thompson et al. 1994). Evolutionary distances were calculated using the Kimura two-parameter model (Kimura 1980). Phylogenetic trees were constructed using the neighbor-joining (NJ) (Saitou and Nei 1987), maximum-likelihood (ML) (Felsenstein 1981), and maximum-parsimony (MP) methods (Fitch 1971), respectively, available in MEGA11 software (Tamura et al. 2021). Bootstrap analysis (1,000 replications) (Felsenstein 1985) was applied to evaluate topologies of NJ, ML, and MP trees.

    Colony morphology of each unreported bacterial species was determined using cells grown on agar plates by culture condition of each strain (Table 1). Cellular morphology and flagellation were examined by transmission electron microscopy (TEM) after uranyl acetate staining of cells grown on agar plates. Gram-staining was performed using a Gram-staining kit (Difco). Biochemical characteristics of unrecorded bacterial strains were tested using API 20NE kits (bioMérieux) according to the manufacturerʹs instructions.

    3. RESULTS AND DISCUSSION

    Through 16S rRNA gene sequence analysis for bacterial isolates obtained from Nakdong River basin, a total of 44 strains were identified as bacterial species unrecorded in Korea. The designation of each strain, ID, 16S rRNA gene sequence similarity (%), and isolation source are described in Table 1. These 44 strains belonged to four bacterial phyla, with twelve strains belonging to phylum Actinomycetota, two strains belonging to phylum Bacillota, six strains belonging to phylum Bacteroidota, and 24 strains belonging to phylum Pseudomonadota (Table 1). The twelve strains belonging to phylum Actinomycetota were found to be grouped with genera Blastococcus, Cellulomonas, Phycicoccus, Klugiella, Subtercola, Paeniglutamicibacter, Mycobacterium, Rhodococcus, Propioniciclava, Raineyella, Nonomuraea, and Actinoallomurus, respectively (Fig. 1). The two strains belonging to phylum Bacillota were grouped with genera Brochothrix and Sporosarcina, respectively (Fig. 1). The six strains belonging to phylum Bacteroidota were found to be grouped to four genera; two strains of Sphingobacterium, two strains of Chryseobacterium, one strain of Flavobacterium, and one strain of Pedobacter, respectively (Fig. 1). The 24 strains belonging to phylum Pseudomonadota were found to be phylogenetically affiliated to thirteen genera Allorhizobium, Endobacterium, Ciceribacter, Rhizobium, Mesorhizobium, Aureimonas, Bauldia, Rhodobacter, Tabrizicola, Methylobacterium, Niveispirillum, Novosphingobium, and Sphingomonas within class α-proteobacteria, six genera Acidovorax, Massilia, Rugamonas, Uruburuella, Methyloversatilis, Zoogloea within class β-proteobacteria, and five genera Rheinheimera, Pantoea, Pseudidiomarina, Pseudomonas, and Stagnimonas within class γ-proteobacteria, respectively (Fig. 2). Among the 44 strains, fifteen strains were short rod-shaped and 27 strains were rod-shaped. Other two strains were filamentous-shaped (Fig. 3). Detailed physiological, morphological, and biochemical characteristics of these 44 bacterial species unrecorded in Korea are given in the strain descriptions. This study expands our knowledge of domestic biodiversity and could contributes to the increase of the industrial application of biological resources.

    3.1. Description of Cellulomonas oligotrophica SL08-109

    Cells are Gram-stain-positive and rod-shaped. Colonies are ivory-pigmented and circular after incubation on 1/10 NA at 25°C for seven days. Cells are positive for nitrate reduction, glucose fermentation, and β-galactosidase and weakly positive for gelatin hydrolysis, but negative for indole production, arginine dihydrolase, urease, esculin hydrolysis, utilization of potassium gluconate, and cytochrome oxidase. d-Glucose, l-arabinose, d-mannose, d-mannitol, N-acetyl-glucosamine, and d-maltose were utilized, but capric acid, adipic acid, malic acid, trisodium citrate, and phenylacetic acid were not utilized as a sole carbon source. Strain SL08-109 (=NNIBR2022641BA2289) was isolated from a freshwater sample collected in Sangju-si, Gyeongsangbukdo, Korea (36°27ʹ16.36ʺN 128°15ʹ31.26ʺE). GenBank accession number for 16S rRNA gene sequence of strain SL08-109 is OQ073730.

    3.2. Description of Blastococcus colisei ND17S-10

    Cells are Gram-stain-positive and short rod-shaped. Colonies are yellow-pigmented, circular, and convex, with entire margin after incubation on R2A at 25°C for seven days. Cells are negative for nitrate reduction, indole production, glucose fermentation, arginine dihydrolase, urease, esculin hydrolysis, gelatin hydrolysis, β-galactosidase, utilization of potassium gluconate, and cytochrome oxidase. d-Glucose, l-arabinose, d-mannose, d-mannitol, N-acetyl-glucosamine, d-maltose, capric acid, adipic acid, malic acid, trisodium citrate, and phenylacetic acid were not utilized. Strain ND17S- 10 (=NNIBR2022641BA2267) was isolated from a freshwater sediment sample collected in Gyeongju-si, Gyeongsangbuk-do, Korea (35°51ʹ06.24ʺN 129°18ʹ 33.01ʺE). GenBank accession number for 16S rRNA gene sequence of strain ND17S-10 is OP962418.

    3.3. Description of Phycicoccus flavus ND40W-79

    Cells are Gram-stain-positive and short rod-shaped. Colonies are yellow-pigmented and circular after incubation on R2A at 25°C for seven days. Cells are positive for nitrate reduction, esculin hydrolysis, gelatin hydrolysis, and β-galactosidase, but negative for indole production, glucose fermentation, arginine dihydrolase, urease, utilization of potassium gluconate, and cytochrome oxidase. d-Mannose and d-mannitol were utilized, but d-glucose, l-arabinose, N-acetyl-glucosamine, d-maltose, capric acid, adipic acid, malic acid, trisodium citrate, and phenylacetic acid were not utilized. Strain ND40W-79 (=NNIBR2022641BA2271) was isolated from a freshwater sample collected in Haeundae-gu, Busan, Korea (35°11ʹ26.28ʺN 129°06ʹ10.26ʺE). Gen Bank accession number for 16S rRNA gene sequence of strain ND40W-79 is OP962422.

    3.4. Description of Klugiella xanthotipulae ND36W-16

    Cells are Gram-stain-positive and short rod-shaped. Colonies are white-pigmented, circular, and convex after incubation on R2A at 25°C for seven days. Cells are positive for β-galactosidase, but negative for nitrate reduction, indole production, glucose fermentation, arginine dihydrolase, urease, esculin hydrolysis, gelatin hydrolysis, utilization of potassium gluconate, and cytochrome oxidase. d-Mannose, d-mannitol, and d-maltose were utilized, but d-glucose, l-arabinose, N-acetyl- glucosamine, capric acid, adipic acid, malic acid, trisodium citrate, and phenylacetic acid were not utilized. Strain ND36W-16 (=NNIBR2022641BA2268) was isolated from a freshwater sample collected in Taebaek- si, Gangwon-do, Korea (37°12ʹ01.47ʺN 128°55ʹ 05.69ʺE). GenBank accession number for 16S rRNA gene sequence of strain ND36W-16 is OP962419.

    3.5. Description of Subtercola boreus ND36S-56

    Cells are Gram-stain-positive and short rod-shaped. Colonies are yellow-pigmented, circular, and convex, with entire margin after incubation on R2A at 25°C for seven days. Cells are positive for esculin hydrolysis and β-galactosidase, but negative for nitrate reduction, indole production, glucose fermentation, arginine dihydrolase, urease, gelatin hydrolysis, utilization of potassium gluconate, and cytochrome oxidase. d-Glucose, l-arabinose, d-mannose, d-mannitol, N-acetylglucosamine, d-maltose, capric acid, adipic acid, malic acid, trisodium citrate, and phenylacetic acid were not utilized. Strain ND36S-56 (=NNIBR2022641BA2269) was isolated from a freshwater sediment sample collected in Taebaek-si, Gangwon-do, Korea (37°12ʹ01.47ʺ N 128°55ʹ05.69ʺE). GenBank accession number for 16S rRNA gene sequence of strain ND36S-56 is OP962 420.

    3.6. Description of Paeniglutamicibacter psychrophenolicus 14S-24

    Cells are Gram-stain-negative and irregular short rodshaped. Colonies are cream-colored, circular, smooth, and convex after incubation on R2A at 20°C for three days. Cells are positive for urease, esculin hydrolysis, β-galactosidase, and utilization of potassium gluconate, but negative for nitrate reduction, indole production on tryptophan, glucose fermentation, arginine dihydrolase, gelatin hydrolysis, and cytochrome oxidase. d-Glucose, d-mannose, d-mannitol, d-maltose, adipic acid, malic acid, trisodium citrate, and phenylacetic acid were utilized, but l-arabinose, N-acetyl-glucosamine, and capric acid were not utilized. Strain 14S-24 (=NNIBR2022 641BA547) was isolated from a freshwater sediment sample collected in Hapcheon-gun, Gyeongsangnamdo, Korea (35°30ʹ37.55ʺN 128°03ʹ16.88ʺE). GenBank accession number for 16S rRNA gene sequence of strain 14S-24 is OP872600.

    3.7. Description of Mycobacterium iranicum 21S-146

    Cells are Gram-stain-positive and rod-shaped. Colonies are yellow-colored, circular, smooth, and convex after incubation on R2A at 20°C for three days. Cells are positive for urease, but negative for nitrate reduction, indole production, glucose fermentation, arginine dihydrolase, esculin hydrolysis, gelatin hydrolysis, β-galactosidase, utilization of potassium gluconate, and cytochrome oxidase. d-Mannitol and malic acid were weakly utilized, but d-glucose, l-arabinose, d-mannose, N-acetyl-glucosamine, d-maltose, capric acid, adipic acid, trisodium citrate, and phenylacetic acid were not utilized. Strain 21S-146 (=NNIBR2022641BA1360) was isolated from a freshwater sediment sample collected in Pohang-si, Gyeongsangbuk-do, Korea (36°07ʹ 22.61ʺN 129°20ʹ58.46ʺE). GenBank accession number for 16S rRNA gene sequence of strain 21S-146 is OP872591.

    3.8. Description of Rhodococcus agglutinans 21GGS1-73

    Cells are Gram-stain-positive and short rod-shaped. Colonies are pinkish-beige colored, circular, smooth, and opaque after incubation on R2A at 25°C for three days. Cells are positive for esculin hydrolysis and β-galactosidase, but negative for nitrate reduction, indole production, glucose fermentation, arginine dihydrolase, urease, gelatin hydrolysis, utilization of potassium gluconate, and cytochrome oxidase. d-Glucose, malic acid, and phenylacetic acid were utilized, but l-arabinose, d-mannose, d-mannitol, N-acetyl-glucosamine, d-maltose, capric acid, adipic acid, and trisodium citrate were not utilized. Strain 21GGS1-73 (=NNIBR 2022641BA2279) was isolated from a freshwater sediment sample collected in Sangju-si, Gyeongsangbukdo, Korea (36°30ʹ45.08ʺN 128°09ʹ40.33ʺE). GenBank accession number for 16S rRNA gene sequence of strain 21GGS1-73 is OP999345.

    3.9. Description of Propioniciclava flava 20HJ5-87

    Cells are Gram-stain-negative and rod-shaped. Colonies are yellow-orange pigmented, circular, and convex after incubation on R2A at 25°C for three days. Cells are positive for nitrate reduction, esculin hydrolysis, β-galactosidase, and cytochrome oxidase, but negative for indole production, glucose fermentation, arginine dihydrolase, urease, gelatin hydrolysis, and utilization of potassium gluconate. d-Glucose, d-mannose and dmannitol were utilized, but l-arabinose, N-acetyl-glucosamine, d-maltose, capric acid, adipic acid, malic acid, trisodium citrate and phenylacetic acid were not utilized. Strain 20HJ5-87 (=NNIBR2021641BA1689) was isolated from a freshwater sample collected in Taebaek- si, Gangwon-do, Korea (37°09ʹ34.08ʺN 128°59ʹ 26.09ʺE). GenBank accession number for 16S rRNA gene sequence of strain 20HJ5-87 is OL884215.

    3.10. Description of Raineyella antarctica ND42S-136

    Cells are Gram-stain-positive and short rod-shaped. Colonies are white colored, circular, and convex after incubation on R2A at 25°C for seven days. Cells are positive for esculin hydrolysis, but negative for nitrate reduction, indole production, glucose fermentation, arginine dihydrolase, urease, gelatin hydrolysis, β-galactosidase, utilization of potassium gluconate, and cytochrome oxidase. d-Glucose, l-arabinose, d-mannose, d-mannitol, N-acetyl-glucosamine, d-maltose, capric acid, adipic acid, malic acid, trisodium citrate, and phenylacetic acid were not utilized. Strain ND42S-136 (=NNIBR2022641BA2276) was isolated from a freshwater sediment sample collected in Ulju-gun, Ulsan, Korea (35°25ʹ33.99ʺN 129°18ʹ31.16ʺE). GenBank accession number for 16S rRNA gene sequence of strain ND42S-136 is OP962427.

    3.11. Description of Nonomuraea maritima 21GGS1-PS42

    Cells are Gram-stain-negative and short rod-shaped. Colonies are greyish-beige colored, umbonate, opaque, matt, with undulate margin after incubation on R2A at 25°C for three days. Cells are positive for esculin hydrolysis, gelatin hydrolysis, β-galactosidase, and cytochrome oxidase, but negative for nitrate reduction, indole production, glucose fermentation, arginine dihydrolase, urease, and utilization of potassium gluconate. d-Glucose, l-arabinose, d-mannose, d-maltose, and adipic acid were utilized, but d-mannitol, N-acetylglucosamine, capric acid, malic acid, trisodium citrate, and phenylacetic acid were not utilized. Strain 21GGS1- PS42 (=NNIBR2022641BA2278) was isolated from a freshwater sediment sample collected in Sangju-si, Gyeongsangbuk-do, Korea (36°30ʹ45.08ʺN 128°09ʹ 40.33ʺE). GenBank accession number for 16S rRNA gene sequence of strain 21GGS1-PS42 is OP999342.

    3.12. Description of Actinoallomurus acanthiterrae 21GGS2-50

    Cells are Gram-stain-negative and filamentousshaped. Colonies are creamy-colored, umbonate, and opaque, and consist of substrate mycelium after incubation on R2A at 25°C for three days. Cells are positive for nitrate reduction, esculin hydrolysis, gelatin hydrolysis, β-galactosidase, and utilization of potassium gluconate, but negative for indole production on tryptophan, glucose fermentation, arginine dihydrolase, urease, and cytochrome oxidase. d-Glucose, l-arabinose, d-mannose, d-mannitol, and d-maltose were utilized and N-acetyl-glucosamine and adipic acid were weakly utilized. Capric acid, malic acid, trisodium citrate, and phenylacetic acid were not utilized. Strain 21GGS2-50 (=NNIBR2022641BA2280) was isolated from a freshwater sediment sample collected in Sangju-si, Gyeongsangbuk- do, Korea (36°30ʹ45.08ʺN 128°09ʹ40.33ʺE). GenBank accession number for 16S rRNA gene sequence of strain 21GGS2-50 is OP999346.

    3.13. Description of Brochothrix thermosphacta ND10W-13

    Cells are Gram-stain-positive and filamentousshaped. Colonies are white-colored, circular, convex, with entire margin after incubation on R2A at 25°C for three days. Cells are positive for esculin hydrolysis and utilization of potassium gluconate, but negative for nitrate reduction, indole production, glucose fermentation, arginine dihydrolase, urease, gelatin hydrolysis, β-galactosidase, and cytochrome oxidase. d-Glucose, d-mannose, d-mannitol, N-acetyl-glucosamine, malic acid, and phenylacetic acid were utilized, but l-arabinose, d-maltose, capric acid, adipic acid, and trisodium citrate were not utilized. Strain ND10W-13 (=NNIBR2022641BA2261) was isolated from a freshwater sample collected in Geochang-gun, Gyeongsangnam- do, Korea (35°48ʹ51.19ʺN 127°52ʹ12.83ʺE). Gen Bank accession number for 16S rRNA gene sequence of strain ND10W-13 is OP847233.

    3.14. Description of Sporosarcina thermotolerans 21GGS2-58

    Cells are Gram-stain-positive and rod-shaped. Colonies are light beige-colored, convex, smooth, and translucent after incubation on R2A at 25°C for three days. Cells are positive for nitrate reduction, urease, esculin hydrolysis and gelatin hydrolysis, but negative for indole production, glucose fermentation, arginine dihydrolase, β-galactosidase, utilization of potassium gluconate, and cytochrome oxidase. Trisodium citrate was utilized and d-glucose was weakly utilized. l-Arabinose, d-mannose, d-mannitol, N-acetyl-glucosamine, d-maltose, capric acid, adipic acid, malic acid, and phenylacetic acid were not utilized. Strain 21GGS2-58 (=NNIBR 2022641BA2281) was isolated from a freshwater sediment sample collected in Sangju-si, Gyeongsangbukdo, Korea (36°30ʹ45.08ʺN 128°09ʹ40.33ʺE). GenBank accession number for 16S rRNA gene sequence of strain 21GGS2-58 is OP999347.

    3.15. Description of Chryseobacterium bovis 20HJ5-53

    Cells are Gram-stain-negative and rod-shaped. Colonies are yellow-colored, circular, smooth, opaque, with entire margin after incubation on R2A at 25°C for three days. Cells are positive for esculin hydrolysis, utilization of potassium gluconate, and cytochrome oxidase, but negative for nitrate reduction, indole production, glucose fermentation, arginine dihydrolase, urease, gelatin hydrolysis, and β-galactosidase. d-Glucose, l-arabinose, d-mannitol, adipic acid, and malic acid were utilized, but d-mannose, N-acetyl-glucosamine, d-maltose, capric acid, trisodium citrate, and phenylacetic acid were not utilized. Strain 20HJ5-53 (=NNIBR2021641 BA1688) was isolated from a freshwater sample collected in Taebaek-si, Gangwon-do, Korea (37°09ʹ34.08ʺN 128°59ʹ26.09ʺE). GenBank accession number for 16S rRNA gene sequence of strain 20HJ5-53 is OL884214.

    3.16. Description of Flavobacterium lutivivi ND42W-61

    Cells are Gram-stain-negative and rod-shaped. Colonies are yellow-colored, circular, and convex after incubation on R2A at 25°C for seven days. Cells are positive for esculin hydrolysis, gelatin hydrolysis, and β-galactosidase, but negative for nitrate reduction, indole production, glucose fermentation, arginine dihydrolase, urease, utilization of potassium gluconate, and cytochrome oxidase. d-Glucose, l-arabinose, d-mannose, d-mannitol, N-acetyl-glucosamine, d-maltose, capric acid, adipic acid, malic acid, trisodium citrate, and phenylacetic acid were not utilized. Strain ND42W-61 (=NNIBR2022641BA2270) was isolated from a freshwater sample collected in Ulju-gun, Ulsan, Korea (35° 25ʹ33.99ʺN 129°18ʹ31.16ʺE). GenBank accession number for 16S rRNA gene sequence of strain ND42W-61 is OP962421.

    3.17. Description of Chryseobacterium chaponense 21W-70

    Cells are Gram-stain-negative and rod-shaped. Colonies are yellow-colored, circular, convex, and smooth after incubation on R2A at 20°C for three days. Cells are positive for esculin hydrolysis, gelatin hydrolysis, and cytochrome oxidase, but negative for nitrate reduction, indole production, glucose fermentation, arginine dihydrolase, urease, β-galactosidase, and utilization of potassium gluconate. d-Glucose and d-maltose were utilized and d-mannose, adipic acid, and trisodium citrate were weakly utilized. l-Arabinose, d-mannitol, N-acetyl- glucosamine, capric acid, malic acid, and phenylacetic acid were not utilized. Strain 21W-70 (=NNIBR 2022641BA1386) was isolated from a freshwater sample collected in Pohang-si, Gyeongsangbuk-do, Korea (36°07ʹ22.61ʺN 129°20ʹ58.46ʺE). GenBank accession number for 16S rRNA gene sequence of strain 21W-70 is OL872592.

    3.18. Description of Pedobacter alpinus 22ND02S-062

    Cells are Gram-stain-negative and rod-shaped. Colonies are orange-colored and circular after incubation on R2A at 25°C for seven days. Cells are positive for esculin hydrolysis, β-galactosidase, and cytochrome oxidase, but negative for nitrate reduction, indole production, glucose fermentation, arginine dihydrolase, urease, gelatin hydrolysis, and utilization of potassium gluconate. d-Glucose and N-acetyl-glucosamine were utilized and d-mannose and d-mannitol were weakly utilized. l-Arabinose, d-maltose, capric acid, adipic acid, malic acid, trisodium citrate, and phenylacetic acid were not utilized. Strain 22ND02S-062 (=NNIBR2022 641BA2297) was isolated from a freshwater sediment sample collected in Yecheon-gun, Gyeongsangbuk-do, Korea (36°34ʹ42.05ʺN 128°19ʹ33.75ʺE). GenBank accession number for 16S rRNA gene sequence of strain 22ND02S-062 is OQ073738.

    3.19. Description of Sphingobacterium populi 21TPW-18

    Cells are Gram-stain-positive and rod-shaped. Colonies are white-colored, circular, and convex, with entire edge after incubation on ISP4 at 25°C for seven days. Cells are positive for esculin hydrolysis, β-galactosidase, and utilization of potassium gluconate, but negative for nitrate reduction, indole production, glucose fermentation, arginine dihydrolase, urease, gelatin hydrolysis, and cytochrome oxidase. d-Glucose, l-arabinose, d-mannose, d-mannitol, N-acetyl-glucosamine, and d-maltose were utilized, but capric acid, adipic acid, malic acid, trisodium citrate, and phenylacetic acid were not utilized. Strain 21TPW-18 (=NNIBR2022641 BA2265) was isolated from a freshwater sample collected in Changnyeong-gun, Gyeongsangnam-do, Korea (35°31ʹ46.86ʺN 128°22ʹ55.35ʺE). GenBank accession number for 16S rRNA gene sequence of strain 21TPW- 18 is OP847237.

    3.20. Description of Sphingobacterium endophyticum TPW-55

    Cells are Gram-stain-negative and short rod-shaped. Colonies are ivory-colored and circular after incubation on 1/10 R2A at 25°C for seven days. Cells are positive for glucose fermentation, β-galactosidase, and cytochrome oxidase and weakly positive for indole production, arginine dihydrolase, and gelatin hydrolysis. Cells are negative for nitrate reduction, urease, esculin hydrolysis, and utilization of potassium gluconate. d-Glucose and l-arabinose were utilized and N-acetyl-glucosamine were weakly utilized. d-Mannose, d-mannitol, d-maltose, capric acid, adipic acid, malic acid, trisodium citrate, and phenylacetic acid were not utilized. Strain TPW-55 (=NNIBR2022641BA2293) was isolated from a freshwater sample collected in Changnyeonggun, Gyeongsangnam-do, Korea (35°31ʹ46.86ʺN 128° 22ʹ55.35ʺE). GenBank accession number for 16S rRNA gene sequence of strain TPW-55 is OQ073734.

    3.21. Description of Aureimonas phyllosphaerae TPW-78

    Cells are Gram-stain-negative, flagellated, and rodshaped. Colonies are ivory-colored and circular after incubation on 1/10 NA at 25°C for seven days. Cells are positive for urease, esculin hydrolysis, β-galactosidase, and cytochrome oxidase, but negative for nitrate reduction, indole production, glucose fermentation, arginine dihydrolase, gelatin hydrolysis, and utilization of potassium gluconate. N-Acetyl-glucosamine was weakly utilized, but d-glucose, l-arabinose, d-mannose, d-mannitol, d-maltose, capric acid, adipic acid, malic acid, trisodium citrate, and phenylacetic acid were not utilized. Strain TPW-78 (=NNIBR2022641BA2294) was isolated from a freshwater sample collected in Changnyeonggun, Gyeongsangnam-do, Korea (35°31ʹ46.86ʺN 128° 22ʹ55.35ʺE). GenBank accession number for 16S rRNA gene sequence of strain TPW-78 is OQ073735.

    3.22. Description of Methylobacterium cerastii ND09S-20

    Cells are Gram-stain-positive and rod-shaped. Colonies are pink-colored, circular, and raised, with entire margin after incubation on R2A at 25°C for three days. Cells are positive for nitrate reduction, esculin hydrolysis, β-galactosidase, and cytochrome oxidase, but negative for indole production, glucose fermentation, arginine dihydrolase, urease, gelatin hydrolysis, and utilization of potassium gluconate. d-Glucose, d-mannose, d-mannitol, and phenylacetic acid were utilized, but l-arabinose, N-acetyl-glucosamine, d-maltose, capric acid, adipic acid, malic acid, and trisodium citrate were not utilized. Strain ND09S-20 (=NNIBR2022641BA 2263) was isolated from a freshwater sediment sample collected in Geochang-gun, Gyeongsangnam-do, Korea (35°48ʹ04.05ʺN 127°46ʹ55.34ʺE). GenBank accession number for 16S rRNA gene sequence of strain ND09S- 20 is OP847235.

    3.23. Description of Mesorhizobium sediminum 22ND02W-014

    Cells are Gram-stain-negative and rod-shaped. Colonies are dark ivory-colored and circular after incubation on MA at 25°C for seven days. Cells are positive for cytochrome oxidase and weakly positive for gelatin hydrolysis. Cells are negative for nitrate reduction, indole production, glucose fermentation, arginine dihydrolase, urease, esculin hydrolysis, β-galactosidase, and utilization of potassium gluconate. d-Glucose and dmannose were weakly utilized, but l-arabinose, dmannitol, N-acetyl-glucosamine, d-maltose, capric acid, adipic acid, malic acid, trisodium citrate, and phenylacetic acid were not utilized. Strain 22ND02W-014 (=NNIBR2022641BA2298) was isolated from a freshwater sample collected in Yecheon-gun, Gyeongsangbuk- do, Korea (36°34ʹ42.05ʺN 128°19ʹ33.75ʺE). Gen Bank accession number for 16S rRNA gene sequence of strain 22ND02W-014 is OQ073739.

    3.24. Description of Allorhizobium vitis TPS-035

    Cells are Gram-stain-negative and rod-shaped. Colonies are pale yellow-colored and circular after incubation on NA at 25°C for seven days. Cells are positive for urease, esculin hydrolysis, β-galactosidase, and cytochrome oxidase and weakly positive for utilization of potassium gluconate. Cells are negative for nitrate reduction, indole production, glucose fermentation, arginine dihydrolase, and gelatin hydrolysis. d-Glucose, l-arabinose, d-mannose, d-mannitol, and d-maltose were utilized and N-acetyl-glucosamine, malic acid, and trisodium citrate were weakly utilized. Capric acid, adipic acid and phenylacetic acid were not utilized. Strain TPS-035 (=NNIBR2022641BA2292) was isolated from a freshwater sediment sample in Changnyeong-gun, Gyeongsangnam-do, Korea (35°31ʹ46.86ʺN 128°22ʹ 55.35ʺE). GenBank accession number for 16S rRNA gene sequence of strain TPS-035 is OQ073733.

    3.25. Description of Bauldia consociata 21GGS1-PS26

    Cells are Gram-stain-negative and short rod-shaped. Colonies are white-colored, circular, convex, and translucent after incubation on R2A at 25°C for three days. Cells are positive for urease, esculin hydrolysis, and cytochrome oxidase and weakly positive for β-galactosidase. Cells are negative for nitrate reduction, indole production, glucose fermentation, arginine dihydrolase, gelatin hydrolysis, and utilization of potassium gluconate. d-Mannitol was utilized and l-arabinose was weakly utilized. d-Glucose, d-mannose, N-acetylglucosamine, d-maltose, capric acid, adipic acid, malic acid, trisodium citrate, and phenylacetic acid were not utilized. Strain 21GGS1-PS26 (=NNIBR2022641BA 2277) was isolated from a freshwater sediment sample collected in Sangju-si, Gyeongsangbuk-do, Korea (36° 30ʹ45.08ʺN 128°09ʹ40.33ʺE). GenBank accession number for 16S rRNA gene sequence of strain 21GGS1-PS 26 is OP999341.

    3.26. Description of Ciceribacter thiooxidans 22M5SE1-20

    Cells are Gram-stain-negative and short rod-shaped. Colonies are white-colored, circular, and convex, entire margin after incubation on R2A at 25°C for seven days. Cells are positive for nitrate reduction, glucose fermentation, urease, esculin hydrolysis, gelatin hydrolysis, β-galactosidase, and cytochrome oxidase, but negative for indole production, arginine dihydrolase, and utilization of potassium gluconate. d-Glucose, l-arabinose, d-mannose, d-mannitol, N-acetyl-glucosamine, d-maltose, malic acid, and trisodium citrate were utilized, but capric acid, adipic acid, and phenylacetic acid were not utilized. Strain 22M5SE1-20 (=NNIBR2022641BA2274) was isolated from a freshwater sediment sample in Sangju-si, Gyeongsangbuk-do, Korea (36°25ʹ07.51ʺN 128°14ʹ36.29ʺE). GenBank accession number for 16S rRNA gene sequence of strain 22M5SE1-20 is OP962 425.

    3.27. Description of Rhizobium subbaraonis 14W-38

    Cells are Gram-stain-positive, flagellated, and rodshaped. Colonies are cream-colored, circular, convex, and smooth after incubation on R2A at 20°C for three days. Cells are positive for urease, esculin hydrolysis, and cytochrome oxidase, but negative for nitrate reduction, indole production, glucose fermentation, arginine dihydrolase, gelatin hydrolysis, β-galactosidase, and utilization of potassium gluconate. d-Glucose, l-arabinose, d-mannitol, and N-acetyl-glucosamine were utilized and d-mannose was weakly utilized. d-Maltose, capric acid, adipic acid, malic acid, trisodium citrate, and phenylacetic acid were not utilized. Strain 14W-38 (=NNIBR 2022641BA579) was isolated from a freshwater sample collected in Hapcheon-gun, Gyeongsangnam-do, Korea (35°30ʹ37.55ʺN 128°03ʹ16.88ʺE). GenBank accession number for 16S rRNA gene sequence of strain 14W-38 is OP872598.

    3.28. Description of Endobacterium yantingense MK07-66

    Cells are Gram-stain-negative and rod-shaped. Colonies are white-colored, circular, and smooth after incubation on MA at 25°C for seven days. Cells are positive for arginine dihydrolase, urease, and cytochrome oxidase, but negative for nitrate reduction, indole production, glucose fermentation, esculin hydrolysis, gelatin hydrolysis, β-galactosidase, and utilization of potassium gluconate. d-Glucose, l-arabinose, d-mannose, d-mannitol, and N-acetyl-glucosamine were utilized, but dmaltose, capric acid, adipic acid, malic acid, trisodium citrate, and phenylacetic acid were not utilized. Strain MK07-66 (=NNIBR2022641BA2290) was isolated from a freshwater sample collected in Gimhae-si, Gyeongsangnam- do, Korea (35°17ʹ20.64ʺN 128°59ʹ42.98ʺE). GenBank accession number for 16S rRNA gene sequence of strain MK07-66 is OQ073731.

    3.29. Description of Rhodobacter thermarum 22ND02S-050

    Cells are Gram-stain-negative and rod-shaped. Colonies are bright red-colored and circular after incubation on R2A at 25°C for seven days. Cells are positive for glucose fermentation, β-galactosidase, and cytochrome oxidase, but negative for nitrate reduction, indole production, arginine dihydrolase, urease, esculin hydrolysis, gelatin hydrolysis, and utilization of potassium gluconate. d-Glucose and d-mannitol were utilized, but larabinose, d-mannose, N-acetyl-glucosamine, capric acid, adipic acid, malic acid, trisodium citrate, and phenylacetic acid were not utilized. Strain 22ND02S- 050 (=NNIBR2022641BA2296) was isolated from a fresh-water sediment sample collected in Yecheon-gun, Gyeongsangbuk-do, Korea (36°34ʹ42.05ʺN 128°19ʹ 33.75ʺE). GenBank accession number for 16S rRNA gene sequence of strain 22ND02S-050 is OQ073737.

    3.30. Description of Tabrizicola aquatica 01TYR-1-41

    Cells are Gram-stain-negative and rod-shaped. Colonies are pink-colored, circular, convex, and smooth, with entire edge after incubation on 1/10 TSA at 25°C for four days. Cells are positive for urease, esculin hydrolysis, β-galactosidase, and cytochrome oxidase, but negative for nitrate reduction, indole production, glucose fermentation, arginine dihydrolase, gelatin hydrolysis, and utilization of potassium gluconate. d-Glucose, d-mannose, d-mannitol, N-acetyl-glucosamine, and adipic acid were utilized, but l-arabinose, d-maltose, capric acid, malic acid, trisodium citrate, and phenylacetic acid were not utilized. Strain 01TYR-1-41 (=NNIBR2020641BA783) was isolated from a freshwater sample collected in Geoje-si, Gyeongsangnamdo, Korea (34°54ʹ56.79ʺN 128°39ʹ18.06ʺE). GenBank accession number for 16S rRNA gene sequence of strain 01TYR-1-41 is MW367776.

    3.31. Description of Niveispirillum cyanobacteriorum 22M6SE1-9

    Cells are Gram-stain-negative and short rod-shaped. Colonies are white-colored, circular, and convex, with entire margin after incubation on R2A at 25°C for seven days. Cells are positive for nitrate reduction, esculin hydrolysis, β-galactosidase, and cytochrome oxidase, but negative for indole production, glucose fermentation, arginine dihydrolase, urease, gelatin hydrolysis, and utilization of potassium gluconate. d-Glucose, larabinose, d-mannose, d-maltose, adipic acid, and trisodium citrate were utilized, but d-mannitol, N-acetylglucosamine, capric acid, malic acid, and phenylacetic acid were not utilized. Strain 22M6SE1-9 (=NNIBR 2022641BA2275) was isolated from a freshwater sediment sample collected in Sangju-si, Gyeongsangbukdo, Korea (36°25ʹ07.51ʺN 128°14ʹ36.29ʺE). GenBank accession number for 16S rRNA gene sequence of strain 22M6SE1-9 is OP962426.

    3.32. Description of Novosphingobium hassiacum 14W-109

    Cells are Gram-stain-negative and rod-shaped. Colonies are yellow-colored, circular, convex, and smooth after incubation on R2A at 20°C for three days. Cells are positive for nitrate reduction, esculin hydrolysis, and β-galactosidase and weakly positive for utilization of potassium gluconate. Cells are negative for indole production, glucose fermentation, arginine dihydrolase, urease, gelatin hydrolysis, and cytochrome oxidase. d-Glucose and malic acid were utilized and d-mannose and adipic acid were weakly utilized. l-Arabinose, d-mannitol, N-acetyl-glucosamine, d-maltose, capric acid, trisodium citrate, and phenylacetic acid were not utilized. Strain 14W-109 (=NNIBR2022641BA592) was isolated from a freshwater sample collected in Hapcheon- gun, Gyeongsangnam-do, Korea (35°30ʹ37.55ʺN 128°03ʹ16.88ʺE). GenBank accession number for 16S rRNA gene sequence of strain 14W-109 is OP872599.

    3.33. Description of Sphingomonas oligophenolica ND29W-66

    Cells are Gram-stain-negative and rod-shaped. Colonies are yellow-colored, circular, convex, and smooth after incubation on R2A at 30°C for three days. Cells are positive for esculin hydrolysis, β-galactosidase, utilization of potassium gluconate, and cytochrome oxidase, but negative for nitrate reduction, indole production, glucose fermentation, arginine dihydrolase, urease, and gelatin hydrolysis. d-Glucose, l-arabinose, d-mannitol, and malic acid were utilized, but d-mannose, N-acetylglucosamine, d-maltose, capric acid, adipic acid, trisodium citrate, and phenylacetic acid were not utilized. Strain ND29W-66 (=NNIBR2022641BA1119) was isolated from a freshwater sample collected in Uljin-gun, Gyeongsangbuk-do, Korea (36°43ʹ11.32ʺN 129°23ʹ 45.85ʺE). GenBank accession number for 16S rRNA gene sequence of strain ND29W-66 is OQ073795.

    3.34. Description of Acidovorax caeni 20HJ5-23

    Cells are Gram-stain-negative and rod-shaped. Colonies are yellow-brown colored, circular, and smooth after incubation on R2A at 25°C for three days. Cells are positive for nitrate reduction, indole production, and cytochrome oxidase, but negative for glucose fermentation, arginine dihydrolase, urease, esculin hydrolysis, gelatin hydrolysis, β-galactosidase, and utilization of potassium gluconate. Malic acid was utilized, but dglucose, l-arabinose, d-mannose, d-mannitol, N-acetyl- glucosamine, d-maltose, capric acid, adipic acid, trisodium citrate, and phenylacetic acid were not utilized. Strain 20HJ5-23 (=NNIBR2021641BA1687) was isolated from a freshwater sample collected in Taebaeksi, Gangwon-do, Korea (37°09ʹ34.08ʺN 128°59ʹ26.09ʺ E). GenBank accession number for 16S rRNA gene sequence of strain 20HJ5-23 is OL884213.

    3.35. Description of Massilia agri 21TPS-072

    Cells are Gram-stain-positive and short rod-shaped. Colonies are yellow-colored, circular, and convex, with entire margin after incubation on R2A at 25°C for three days. Cells are positive for cytochrome oxidase, but negative for nitrate reduction, indole production, glucose fermentation, arginine dihydrolase, urease, esculin hydrolysis, gelatin hydrolysis, β-galactosidase, and utilization of potassium gluconate. l-arabinose, d-maltose, and malic acid were weakly utilized, but d-glucose, d-mannose, d-mannitol, N-acetyl-glucosamine, capric acid, adipic acid, trisodium citrate, and phenylacetic acid were not utilized. Strain 21TPS-072 (=NNIBR2022641BA2264) was isolated from a freshwater sediment sample collected in Changnyeong-gun, Gyeongsangnam-do, Korea (35°31ʹ46.86ʺN 128°22ʹ 55.35ʺE). GenBank accession number for 16S rRNA gene sequence of strain 20HJ5-23 is OP847236.

    3.36. Description of Rugamonas rivuli ND10S-28

    Cells are Gram-stain-positive and short rod-shaped. Colonies are gray-colored, circular, and convex, with entire margin after incubation on R2A at 25°C for three days. Cells are positive for nitrate reduction and esculin hydrolysis, but negative for indole production, glucose fermentation, arginine dihydrolase, urease, gelatin hydrolysis, β-galactosidase, utilization of potassium gluconate, and cytochrome oxidase. l-Arabinose, d-mannose, and d-maltose were utilized, but d-glucose, d-mannitol, N-acetyl-glucosamine, capric acid, adipic acid, malic acid, trisodium citrate, and phenylacetic acid were not utilized. Strain ND10S-28 (=NNIBR2022 641BA2259) was isolated from a freshwater sediment sample collected in Geochang-gun, Gyeongsangnamdo, Korea (35°48ʹ51.19ʺN 127°52ʹ12.83ʺE). GenBank accession number for 16S rRNA gene sequence of strain ND10S-28 is OP847231.

    3.37. Description of Uruburuella suis 20HJ5-7

    Cells are Gram-stain-negative and rod-shaped. Colonies are yellow-colored, circular, and smooth, with entire margin after incubation on R2A at 25°C for three days. Cells are positive for urease, utilization of potassium gluconate, and cytochrome oxidase, but negative for nitrate reduction, indole production, glucose fermentation, arginine dihydrolase, esculin hydrolysis, gelatin hydrolysis, and β-galactosidase. d-Glucose, l-arabinose, and malic acid were utilized and d-mannose was weakly utilized. d-mannitol, N-acetyl-glucosamine, d-maltose, capric acid, adipic acid, trisodium citrate, and phenylacetic acid were not utilized. Strain 20HJ5- 7 (=NNIBR2021641BA1686) was isolated from a freshwater sample collected in Taebaek-si, Gangwon-do, Korea (37°09ʹ34.08ʺN 128°59ʹ26.09ʺE). GenBank accession number for 16S rRNA gene sequence of strain 20HJ5-7 is OL884212.

    3.38. Description of Methyloversatilis universalis 22M5SE1-5

    Cells are Gram-stain-negative and short rod-shaped. Colonies are white-colored, circular, and convex after incubation on R2A at 25°C for seven days. Cells are positive for nitrate reduction, esculin hydrolysis, and cytochrome oxidase, but negative for indole production, glucose fermentation, arginine dihydrolase, urease, gelatin hydrolysis, β-galactosidase, and utilization of potassium gluconate. Malic acid was utilized, but dglucose, l-arabinose, d-mannose, d-mannitol, N-acetyl- glucosamine, d-maltose, capric acid, adipic acid, trisodium citrate, and phenylacetic acid were not utilized. Strain 22M5SE1-5 (=NNIBR2022641BA2273) was isolated from a freshwater sediment sample collected in Sangju-si, Gyeongsangbuk-do, Korea (36°25ʹ 07.51ʺN 128°14ʹ36.29ʺE). GenBank accession number for 16S rRNA gene sequence of strain 22M5SE1-5 is OP962424.

    3.39. Description of Zoogloea oryzae 22ND02S-042

    Cells are Gram-stain-negative and rod-shaped. Colonies are pale ivory-colored and circular after incubation on R2A at 25°C for seven days. Cells are positive for nitrate reduction, urease, utilization of potassium gluconate, and cytochrome oxidase, but negative for indole production, glucose fermentation, arginine dihydrolase, esculin hydrolysis, gelatin hydrolysis, and β-galactosidase. d-Glucose, l-arabinose, d-mannose, d-mannitol, N-acetyl-glucosamine, d-maltose, and capric acid were utilized, but adipic acid, malic acid, trisodium citrate, and phenylacetic acid were not utilized. Strain 22ND02S-042 (=NNIBR2022641BA2295) was isolated from a freshwater sediment sample collected in Yecheon- gun, Gyeongsangbuk-do, Korea (36°34ʹ42.05ʺN 128°19ʹ33.75ʺE). GenBank accession number for 16S rRNA gene sequence of strain 22ND02S-042 is OQ073 736.

    3.40. Description of Pseudidiomarina gelatinasegens 20HJ-1-18

    Cells are Gram-stain-negative and rod-shaped. Colonies are light yellow-colored, circular, convex, and transparent, with entire margins after incubation on MA at 25°C for three days. Cells are positive for indole production and cytochrome oxidase, but negative for nitrate reduction, glucose fermentation, arginine dihydrolase, urease, esculin hydrolysis, gelatin hydrolysis, β-galactosidase, and utilization of potassium gluconate. d-Mannose and d-mannitol were utilized, but dglucose, l-arabinose, N-acetyl-glucosamine, d-maltose, capric acid, adipic acid, malic acid, trisodium citrate, and phenylacetic acid were not utilized. Strain 20HJ- 1-18 (=NNIBR2021641BA1685) was isolated from a freshwater sample collected in Taebaek-si, Gangwondo, Korea (37°12ʹ28.06ʺN 128°55ʹ54.04ʺE). GenBank accession number for 16S rRNA gene sequence of strain 20HJ-1-18 is OL884211.

    3.41. Description of Rheinheimera nanhaiensis 22ND44W-039

    Cells are Gram-stain-negative and rod-shaped. Colonies are ivory-colored and irregular after incubation on 1/10 TSA at 25°C for seven days. Cells are positive for glucose fermentation, esculin hydrolysis, gelatin hydrolysis, and cytochrome oxidase, but negative for nitrate reduction, indole production, arginine dihydrolase, urease, β-galactosidase, and utilization of potassium gluconate. d-Glucose, l-arabinose, d-mannose, d-mannitol, N-acetyl-glucosamine, d-maltose, capric acid, adipic acid, malic acid, trisodium citrate, and phenylacetic acid were utilized. Strain 22ND44W-039 (=NNIBR 2022641BA2317) was isolated from a freshwater sample collected in Ulju-gun, Ulsan, Korea (35°27ʹ32.91ʺN 129°17ʹ28.35ʺE). GenBank accession number for 16S rRNA gene sequence of strain 20HJ-1-18 is OQ073744.

    3.42. Description of Pantoea cypripedii 21TPW-40

    Cells are Gram-stain-positive and rod-shaped. Colonies are white-colored, circular, and convex, with entire margin after incubation on R2A at 25°C for three days. Cells are positive for nitrate reduction, esculin hydrolysis, β-galactosidase, and utilization of potassium gluconate, but negative for indole production, glucose fermentation, arginine dihydrolase, urease, gelatin hydrolysis, and cytochrome oxidase. d-Glucose, l-arabinose, d-mannose, d-mannitol, N-acetyl-glucosamine, dmaltose, malic acid, and trisodium citrate were utilized, but capric acid, adipic acid, and phenylacetic acid were not utilized. Strain 21TPW-40 (=NNIBR2022641BA 2266) was isolated from a freshwater sample collected in Changnyeong-gun, Gyeongsangnam-do, Korea (35° 31ʹ46.86ʺN 128°22ʹ55.35ʺE). GenBank accession number for 16S rRNA gene sequence of strain 21TPW-40 is OP847238.

    3.43. Description of Pseudomonas akapageensis ND10S-45

    Cells are Gram-stain-positive and rod-shaped. Colonies are white-colored, circular, and raised after incubation on R2A at 25°C for three days. Cells are positive for nitrate reduction, arginine dihydrolase, urease, utilization of potassium gluconate, and cytochrome oxidase and weakly positive for esculin hydrolysis. Cells are negative for indole production, glucose fermentation, gelatin hydrolysis, and β-galactosidase. d-Glucose, d-mannose, malic acid, trisodium citrate, and phenylacetic acid were utilized, but l-arabinose, d-mannitol, N-acetyl-glucosamine, d-maltose, capric acid, and adipic acid were not utilized. Strain ND10S-45 (=NNIBR2022641BA2260) was isolated from a freshwater sediment sample collected in Geochang-gun, Gyeongsangnam-do, Korea (35°48ʹ51.19ʺN 127°52ʹ 12.83ʺE). GenBank accession number for 16S rRNA gene sequence of strain ND10S-45 is OP847232.

    3.44. Description of Stagnimonas aquatica 22M6S-19

    Cells are Gram-stain-negative and rod-shaped. Colonies are white-colored, circular, and convex after incubation on R2A at 25°C for seven days. Cells are positive for esculin hydrolysis, gelatin hydrolysis, utilization of potassium gluconate, and cytochrome oxidase, but negative for nitrate reduction, indole production, glucose fermentation, arginine dihydrolase, urease, and β-galactosidase. d-Glucose, l-arabinose, d-mannose, d-mannitol, N-acetyl-glucosamine, d-maltose, capric acid, adipic acid, malic acid, trisodium citrate, and phenylacetic acid were not utilized. Strain 22M6S-19 (=NNIBR 2022641BA2272) was isolated from a freshwater sediment sample collected in Sangju-si, Gyeongsangbuk- do, Korea (36°25ʹ07.51ʺN 128°14ʹ36.29ʺE). Gen Bank accession number for 16S rRNA gene sequence of strain 22M6S-19 is OP962423.

    ACKNOWLEDGEMENTS

    This work was supported by a Nakdonggang National Institute of Biological Resources grant (project no. NNIBR202301103) funded by the Ministry of Environment, Republic of Korea.

    CRediT authorship contribution statement

    JH Jeon: Investigation, Writing - Original draft. S Park, JY Cho, and SY Lee: Resource, Methodology, Investigation, Data curation. S Hwang and JS Kim: Investigation. EJ Kim: Project administration, Writing - Review and editing. JY Jung: Supervision, Writing - Original draft, review, and editing.

    Declaration of Competing Interest

    The authors declare no conflicts of interest.

    Figure

    KJEB-41-3-308_F1.gif

    A neighbor-joining phylogenetic tree, based on 16S rRNA gene sequences, showing the relationship between twenty strains belonging to the phyla Actinomycetota, Bacillota, and Bacteroidota. Bootstrap values (expressed as percentages of 1,000 replications) above 50% are shown at branch points. Bar, 0.05 substitutions per nucleotide position. Filled and empty circles indicate nodes recovered by all three and two algorithms (neighbor-joining, maximum likelihood, and maximum parsimony), respectively. Escherichia coli ATCC 11775T (X80725) was used as an outgroup.

    KJEB-41-3-308_F2.gif

    A neighbor-joining phylogenetic tree, based on 16S rRNA gene sequences, showing the relationship between 24 strains belonging to the Phylum Pseudomonadota. Bootstrap values (expressed as percentages of 1,000 replications) above 50% are shown at branch points. Bar, 0.05 substitutions per nucleotide position. Filled and empty circles indicate nodes recovered by all three and two algorithms (neighbor-joining, maximum likelihood, and maximum parsimony), respectively. Flavobacterium circumlabens CCM 8828T (MH100898) was used as an outgroup.

    KJEB-41-3-308_F3.gif

    Transmission electron or scanning electron micrographs of cells of strains isolated in the study. Strains: 1, SL08 -109; 2, ND17S-10; 3, ND40W-79; 4, ND36W-16; 5, ND36S-56; 6, 14S-24; 7, 21S-146; 8, 21GGS1-73; 9, 20HJ5-87; 10, ND42S-136; 11, 21GGS1-PS42; 12, 21GGS2-50; 13, ND10W-13; 14, 21GGS2-58; 15, 20HJ5-53; 16, ND42W-61; 17, 21W-70; 18, 22ND02S-062; 19, 21TPW-18; 20, TPW-55; 21, TPW-78; 22, ND09S-20; 23, 22ND02W-014; 24, TPS-035; 25, 21GGS1-PS26; 26, 22M5SE1-20; 27, 14W-38; 28, MK07-66; 29, 22ND02S-050; 30, 01TYR-1-41; 31, 22M6SE1-9; 32, 14W-109; 33, ND29W-66; 34, 20HJ5-23; 35, 21TPS-072; 36, ND10S-28; 37, 20HJ5-7; 38, 22M5SE1-5; 39, 22ND02S-042; 40, 20HJ-1-18; 41, 22ND44W-039; 42, 21TPW-40; 43, ND10S-45; 44, 22M6S-19.

    Table

    A summary of the 44 unrecorded bacterial strains and taxonomic affiliations based on 16S rRNA gene sequence

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    Vol. 40 No. 4 (2022.12)

    Journal Abbreviation 'Korean J. Environ. Biol.'
    Frequency quarterly
    Doi Prefix 10.11626/KJEB.
    Year of Launching 1983
    Publisher Korean Society of Environmental Biology
    Indexed/Tracked/Covered By

    Contact info

    Any inquiries concerning Journal (all manuscripts, reviews, and notes) should be addressed to the managing editor of the Korean Society of Environmental Biology. Yongeun Kim,
    Korea University, Seoul 02841, Korea.
    E-mail: kyezzz@korea.ac.kr /
    Tel: +82-2-3290-3496 / +82-10-9516-1611