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ISSN : 1226-9999(Print)
ISSN : 2287-7851(Online)
Korean J. Environ. Biol. Vol.41 No.3 pp.215-222
DOI : https://doi.org/10.11626/KJEB.2023.41.3.215

A report of 10 unrecorded bacterial species of Korea, isolated from agricultural soil in 2022

Oung Bin Lim, Ji Soo Lee, Hyosun Lee, Ki Eun Lee1, In Tae Cha1, Won Jae Chi1, Dong-Uk Kim*
Department of Biological Science, College of Biological and Environmental Sciences, Sangji University, Wonju 26339, Republic of Korea
1Microorganism Resources Division, National Institute of Biological Resources, Incheon 22689, Republic of Korea
* Corresponding author Dong-Uk Kim Tel. 033-730-0433 E-mail. dukim@sangji.ac.kr

Contribution to Environmental Biology


▪ Since the Nagoya Protocol was issued, the value of biological resources has been increasing.


▪ In this study, we would like to report a taxonomic aspect of unrecorded species that have been reported abroad but not yet reported in Korea.


▪ The unrecorded species that were discovered are expected to be valuable for use as biological and genetic resources.

11/05/2023 21/08/2023 24/08/2023

Abstract


In 2022, research for native prokaryotic species in Korea reported 10 unrecorded bacterial strains affiliated to phyla Actinomycetota, Bacillota, and Pseudomonadota. The strains formed monophyletic clades with the most closely related species (with ≥98.7% sequence similarity) in the 16S rRNA gene sequencing. Among them, four species of the phylum Actinomycetota, two species of the phylum Bacillota, and four species of the phylum Pseudomonadota have not been reported in Korea, suggesting unrecorded species in Korea. Information on strains such as Gram staining reaction, colony and cell morphology, biochemical characteristics, and isolation sources were provided in the species description.


16S rRNA , bacterial diversity , unrecorded species

초록

    1. INTRODUCTION

    For the discover indigenous prokaryotic species in Korea, various environmental samples are collected (Jang et al. 2022). In 2022, 10 unrecorded bacteria were isolated from agricultural soil samples from various regions in Korea and identified as phyla of Actinomycetes, Bacillota and Pseudomonadota. The taxonomic information of 10 unrecorded species is based on the nomenclature described in List of Prokaryotic names with Standing in Nomenclature (https://lpsn.dsmz.de) and consists of Actinomycetota, Bacillota and Pseudomonadota. The phylum Actinomycetota, one of the largest phyla within the domain Bacteria, is widely distributed in aquatic and terrestrial environments (Lawson 2018). The phylum is comprised of Gram stain positive, non-sporeforming, rod-shaped bacteria with a high G+C content (Gao and Gupta 2005); the members of the phylum Bacillota, which are resistant to desiccation and can survive extreme conditions and distributed in diverse environments, are known as Gram stain positive and low G+C content containing rod/coccusshaped bacteria (Nahar et al. 2018); the phylum Pseudomonadota are known as Gram stain negative bacteria and responsible for nitrogen fixation, which one of the largest phyla within the domain Bacteria (Seong et al. 2019).

    This report focuses on the description of the charac-teristics that 10 unrecorded species belonging to the phyla Actinomycetota, Bacillota, and Pseudomonadota, which have not been reported in Korea so far.

    2. MATERIALS AND METHODS

    A total of 10 bacterial strains assigned to the phyla Actinomycetota, Bacillota and Pseudomonadota were isolated from agricultural soil. The agricultural soil samples collected from each region were independently processed serial dilution and spread onto diverse culture media Reasoner’2A agar (R2A; BD Difco), nutrient agar (NA; BD Difco) and tryptic soy agar (TSA; BD Difco) and incubated at 28°C for 3-7 days (Table 1). All strains were purified as single colonies and stored as 15-17% glycerol suspension at -80°C as well as lyophilized ampoules.

    The morphology and cell size of colonies for each strain were determined by using a scanning electron microscopy and transmission electron microscopy. The electron micrographs of each strain are shown in Figure 1. Gram reaction was performed according to the classic Gram procedure described by Doetsch (1981). The biochemical characteristics of the strains were tested using the API test kit (API 20NE, API 32GN, API ZYM) by referring to the bioMérieux company’s instructions. After extracting genomic DNA and amplify the 16S rRNA gene using 27mF, 1492R (universal) primers (Han et al. 2022), and the sequence database of each strain was submission in NCBI GenBank. The 16S rRNA gene sequences of the related taxa were obtained from EzBioCloud server (Yoon et al. 2017). Ten bacterial strains and related taxa (retrieved from the NCBI database) were aligned with SINA (v1.2.12) according to the SILVA seed alignment (http://www.arb-silva. de; Pruesse et al. 2012). Evolutionary distances applied during phylogenetic analysis were calculated through a two-parameter model (Kimura 1983). A phylogenetic tree was constructed using bootstrap values (Felsenstein 1985) based on 1,000 replicates using neighbor joining (Saitou and Nei 1987) in the MEGA X program (Kumar et al. 2018).

    3. RESULTS AND DISCUSSION

    The 10 strains were distributed into 3 phyla: Actinomycetota, Bacillota, Pseudomonadota (Table 1). All strains were rod-shaped (Fig. 1). Unrecorded bacteria were identified as 10 genera of Afipia, Aquamicrobium, Corynebacterium, Gordonia, Intrasporangium, Mesorhizobium, Mycoplana, Nocardioides and Paenibacillus (Fig. 2). Here we report 10 unrecorded bacterial species in Korea belonging to 5 orders, which were isolated in Korea; 4 strains of the Hyphomicrobiales, 1 strain of the Micrococcales, 2 strains of the Mycobacteriales, 2 strains of the Paenibacillaceae, and 1 strain of the Propionibacteriales.

    3.1. Description of Nocardioides jensenii GW31

    Cells are Gram-staining-positive and rod-shape. Cell size is 1.2-2.2 μm. Colonies are circular, smooth, entire and white color after 4 days of incubation at 28°C on NA. In API 20NE, the reactions in nitrate reduction (NO3- → NO2-), and utilization of arginine dihydrolase, urease, D-glucose, gluconate, adipate, and malate are positive, but the reaction in reduction of indole production, glucose acidification, β-glucosidase (esculin hydrolysis), protease (gelatin hydrolysis), β-galactosidase (PNPG), L-arabinose, D-mannose, D-mannitol, N-acetyl-D-glucosamine, D-maltose, caprate, and citrate are negative. In API 32GN, the reaction in D-glucose, and utilization of L-fucose, D-sorbitol, valerate, L-histidine, 3-hydroxy-butyrate, L-rhamnose, L-alanine glycogen, and 3-hydroxy-benzoate are positive, but reaction in D-mannitol, salicin, D-melibiose, L-arabinose, propionate, caprate, citrate, 2-ketogluconate, 4- hydroxy-benzoate, L-proline, N-acetyl-D-glucosamine, D-ribose, inositol, D-sucrose, D-maltose, itaconate, suberate, malonate, acetate, lactate, 5-ketogluconate, and L-serine are negative. Positive reactions for enzymatic activity in alkaline phosphatase, esterase (C4), esterase lipase (C8), leucine arylamidase, acid phosphatase, naphtol-AS-BI-phosphohydrolase, α-glucosidase, and α-mannosidase at API ZYM, but negative reactions for lipase (C14), valine arylamidase, cystine arylamidase, trypsin, α-chymotrypsin, α-galactosidase, β-galactosidase, β-glucuronidase, β-glucosidase, N-acetyl- β-glucosaminidase, and α-fucosidase. Strain GW31 (=NIBRBAC000509779) was isolated an agricultural soil sample, Gyeongsangnam-do, Korea (35°23ʹ43.2ʺN 128°08ʹ54.3ʺE).

    3.2. Description of Gordonia bronchialis R6

    Cells are Gram-staining-positive and rod-shape. Cell size is 1.5-2.2 μm. Colonies are circular, smooth, entire and orange color after 4 days of incubation at 28°C on NA. In API 20NE, the reactions in nitrate reduction (NO3- → NO2-), and utilization of D-glucose, D-mannose, D-mannitol, N-acetyl-D-glucosamine, and malate are positive, but the reaction in indole production, glucose acidification, arginine dihydrolase, protease (gelatin hydrolysis), β-galactosidase (PNPG), L-arabinose, D-maltose, gluconate, caprate, citrate, phenylacetate. In API 32GN, the reaction in D-mannitol, and utilization of D-glucose, propionate, valerate, L-histidine, 3-hydroxy-butyrate, L-proline, L-rhamnose, Nacetyl- D-glucosamine, inositol, acetate, and lactate are positive, but reaction in salicin, D-melibiose, L-fucose, D-sorbitol, L-arabinose, caprate, citrate, 2-ketogluconate, 4-hydroxy-benzoate, D-ribose, D-sucrose, D-maltose, itaconate, suberate, malonate, L-alanine, 5-ketogluconate, glycogen, 3-hydroxy-benzoate, and L-serine are negative. Positive reactions for enzymatic activity in alkaline phosphatase, esterase (C4), esterase lipase (C8), leucine arylamidase, cystine arylamidase, acid phosphatase, naphtol-AS-BI-phosphohydrolase, α-glucosidase, and β-glucosidase at API ZYM, but negative reactions for lipase (C14), valine arylamidase, trypsin, α-chymotrypsin, α-galactosidase, β-galactosidase, β-glucuronidase, N-acetyl-β-glucosaminidase, α-mannosidase, and α-fucosidase. Strain R6 (=NIBR BAC000509780) was isolated from an agricultural soil sample, Chungcheongnam-do, Korea (36°53ʹ11.8ʺN 126°44ʹ34.7ʺE).

    3.3. Description of Corynebacterium glucuronolyticum JN1306

    Cells are Gram-staining-positive and rod-shape. Cell size is 1.2-1.5 μm. Colonies are circular, smooth, entire and white color after 3 days of incubation at 28°C on NA. In API 20NE, the reactions in nitrate reduction (NO3- → NO2-), and utilization of D-glucose are positive, but the reaction in indole production, glucose acidification, arginine dihydrolase, urease, β-glucosidase (esculin hydrolysis), protease (gelatin hydrolysis), β-galactosidase (PNPG), L-arabinose, D-mannose, Dmannitol, N-acetyl-D-glucosamine, D-maltose, gluconate, caprate, adipate, malate, citrate, and phenylacetate are negative. In API 32GN, the reaction in Dglucose, and utilization of L-fucose, propionate, D-sucrose, and glycogen are positive, but reaction in D-mannitol, salicin, D-melibiose, D-sorbitol, L-arabinose, caprate, valerate, citrate, L-histidine, 2-ketogluconate, 3-hydroxy-butyrate, 4-hydroxy-benzoate, L-proline, L-rhamnose, N-acetyl-D-glucosamine, D-ribose, inositol, D-maltose, itaconate, suberate, malonate, acetate, lactate, L-alanine, 5-ketogluconate, 3-hydroxy-benzoate, and L-serine are negative. Positive reactions for enzymatic activity in esterase (C4), esterase lipase (C8), leucine arylamidase, acid phosphatase, naphtol-AS-BIphosphohydrolase, and β-glucuronidase at API ZYM, but negative reactions for alkaline phosphatase, lipase (C14), valine arylamidase, cystine arylamidase, trypsin, α-chymotrypsin, α-galactosidase, β-galactosidase, α-glucosidase, β-glucosidase, N-acetyl-β-glucosaminidase, α-mannosidase, and α-fucosidase. Strain JN1306 (=NIBRBAC000509785) was isolated from an agricultural sample, Jeollabuk-do, Korea (35°33ʹ02.5ʺN 127°08ʹ 14.6ʺE).

    3.4. Description of Intrasporangium chromatireducens SW56

    Cells are Gram-staining-positive and rod-shape. Cell size is 1.0-1.7 μm. Colonies are circular, smooth, entire and white color after 4 days of incubation at 28°C on TSA. In API 20NE, the reactions in nitrate reduction (NO3- → NO2-), and utilization of urease, β-glucosidase (esculin hydrolysis), protease (gelatin hydrolysis), D-glucose, D-mannose, D-maltose, and gluconate are positive, but the reaction in indole production, glucose acidification, arginine dihydrolase, β-galactosidase (PNPG), L-arabinose, D-mannitol, N-acetyl-D-glucosamine, caprate, adipate, malate, citrate, and phenyl-acetate are negative. In API 32GN, the reaction in D-glucose, and utilization of propionate, valerate, L-histidine, 3-hydroxy-butyrate, L-proline, D-ribose, D-sucrose, Dmaltose, acetate, L-alanine, 3-hydroxy-benzoate, and L-serine are positive, but reaction in D-mannitol, salicin, D-melibiose, L-fucose, D-sorbitol, L-arabinose, caprate, citrate, 2-ketogluconate, 4-hydroxy-benzoate, L-rhamnose, N-acetyl-D-glucosamine, inositol, itaconate, suberate, malonate, lactate, 5-ketogluconate, and glycogen are positive. Positive reactions for enzymatic activity in esterase (C4), esterase lipase (C8), leucine arylamidase, acid phosphatase, naphtol-AS-BI-phosphohydrolase, β-glucuronidase, and α-glucosidase at API ZYM, but negative reactions for alkaline phosphatase, lipase (C14), valine arylamidase, cystine arylamidase, trypsin, α-chymotrypsin, α-galactosidase, β-galactosidase, β-glucosidase, N-acetyl-β-glucosaminidase, α-mannosidase, and α-fucosidase. Strain SW56 (=NIBRBAC000509786) was isolated from an agricultural soil sample, Chungcheongbuk-do, Korea (36°47ʹ 30.5ʺN 127°29ʹ47.1ʺE).

    3.5. Description of Afipia clevelandensis TA-E7

    Cells are Gram-staining-negative and rod-shape. Cell size is 1.1-1.6 μm. Colonies are circular, smooth, entire and white color after 4 days of incubation at 28°C on R2A. In API 20NE, the reactions in reduction of nitrate reduction (NO3- → NO2-), and utilization of arginine dihydrolase, urease, gluconate, adipate, and malate are positive, but the reaction in indole production, glucose acidification, β-glucosidase (esculin hydrolysis), protease (gelatin hydrolysis), β-galactosidase (PNPG), D-glucose, L-arabinose, D-mannose, D-mannitol, N-acetyl- D-glucosamine, D-maltose, caprate, citrate, and phenyl- acetate are negative. In API 32GN, the reaction in propionate, and utilization of valerate, 3-hydroxy-butyrate, itaconate, suberate, acetate, and glycogen are positive, but reaction in D-mannitol, D-glucose, salicin, D-melibiose, L-fucose, D-sorbitol, L-arabinose, caprate, citrate, L-histidine, 2-ketogluconate, 4-hydroxy-benzoate, L-proline, L-rhamnose, N-acetyl-D-glucosamine, D-ribose, inositol, D-sucrose, D-maltose, malonate, lactate, L-alanine, 5-ketogluconate, 3-hydroxy-benzoate, and L-serine are negative. Positive reactions for enzymatic activity in alkaline phosphatase, esterase (C4), esterase lipase (C8), leucine arylamidase, trypsin, and naphtol-AS-BI-phosphohydrolase at API ZYM, but negative reactions for lipase (C14), valine arylamidase, cystine arylamidase, α-chymotrypsin, acid phosphatase, α-galactosidase, β-galactosidase, β-glucuronidase, α-glucosidase, β-glucosidase, N-acetyl-β-glucosaminidase, α-mannosidase, and α-fucosidase. Strain TA-E7 (=NIBRBAC000509781) was isolated from an agricultural soil sample, Gyeonggi-do, Korea (37°06ʹ53.9ʺN 126°58ʹ48.1ʺE).

    3.6. Description of Mesorhizobium thiogangeticum SY19

    Cells are Gram-staining-negative and rod-shape. Cell size is 0.8-1.2 μm. Colonies are circular, smooth, entire and yellow color after 4 days of incubation at 28°C on NA. In API 20NE, the reactions in glucose acidification, and utilization of arginine dihydrolase, urease, β-glucosidase (esculin hydrolysis), D-glucose, L-arabinose, D-mannitol, N-acetyl-D-glucosamine, and gluconate are positive, but the reaction in nitrate reduction (NO3- → NO2-), reduction of nitrates to nitrogen, indole production, protease (gelatin hydrolysis), β-galactosidase (PNPG), D-maltose, caprate, adipate, malate, citrate, and phenyl-acetate are negative. In API 32GN, the reaction in D-mannitol, and utilization of D-glucose, L-fucose, D-sorbitol, L-arabinose, propionate, valerate, L-histidine, 2-ketogluconate, 3-hydroxy-butyrate, 4- hydroxy-benzoate, L-proline, N-acetyl-D-glucosamine, acetate, lactate, L-alanine, and L-serine are positive, but reaction in salicin, D-melibiose, caprate, citrate, L-rhamnose, D-ribose, inositol, D-sucrose, D-maltose, itaconate, suberate, malonate, 5-ketogluconate, and 3-hydroxy- benzoate are negative. Positive reactions for enzymatic activity in alkaline phosphatase, esterase (C4), esterase lipase (C8), leucine arylamidase, valine arylamidase, trypsin, acid phosphatase, and naphtol-ASBI- phosphohydrolase at API ZYM, but negative reactions for lipase (C14), cystine arylamidase, α-chymotrypsin, α-galactosidase, β-galactosidase, β-glucuronidase, α-glucosidase, β-glucosidase, N-acetyl-β-glucosaminidase, α-mannosidase, and α-fucosidase. Strain SY19 (=NIBRBAC000509784) was isolated from an agricultural soil sample, Jeollabuk-do, Korea (35°45ʹ 12.3ʺN 127°08ʹ42.5ʺE).

    3.7. Description of Aquamicrobium soli SY31

    Cells are Gram-staining-negative and rod-shape. Cell size is 0.7-1.1 μm. Colonies are circular, smooth, entire and yellow color after 4 days of incubation at 28°C on TSA. In API 20NE, the reactions in D-glucose, and utilization of L-arabinose, D-mannitol, N-acetyl-D-glucosamine, and gluconate are positive, but the reaction in nitrate reduction (NO3- → NO2-), reduction of nitrates to nitrogen, indole production, arginine dihydrolase, urease, β-glucosidase (esculin hydrolysis), protease (gelatin hydrolysis), β-galactosidase (PNPG), D-mannose, D-maltose, caprate, adipate, malate, citrate, and phenylacetate are negative. In API 32GN, the reaction in Dmannitol, and utilization of D-glucose, L-fucose, D-sorbitol, L-arabinose, propionate, valerate, L-histidine, 3- hydroxy-butyrate, L-proline, L-rhamnose, N-acetyl-Dglucosamine, D-ribose, acetate, lactate, and L-alanine are positive, but reaction in salicin, D-melibiose, caprate, citrate, 2-ketogluconate, 4-hydroxy-benzoate, inositol, D-sucrose, D-maltose, itaconate, suberate, malonate, 5-ketogluconate, glycogen, 3-hydroxy-benzoate, and L-serine are negative. Positive reactions for enzymatic activity in esterase (C4), esterase lipase (C8), leucine arylamidase, acid phosphatase, naphtol-AS-BIphosphohydrolase, α-galactosidase, β-galactosidase, and α-glucosidase at API ZYM alkaline phosphatase, esterase (C4), leucine arylamidase, trypsin, acid phosphatase, and α-glucosidase at API ZYM, but negative reactions for lipase (C14), valine arylamidase, cystine arylamidase, α-chymotrypsin, α-galactosidase, β-galactosidase, β-glucuronidase, β-glucosidase, N-acetyl β-glucosaminidase, α-mannosidase, and α-fucosidase. Strain SY31 (=NIBRBAC000509787) was isolated from an agricultural soil sample, Jeollanam-do, Korea (35°02ʹ 19.3ʺN 127°02ʹ23.5ʺE).

    3.8. Description of Mycoplana dimorpha NOY11-2

    Cells are Gram-staining-negative and rod-shape. Cell size is 1.5-2.4 μm. Colonies are irregular, smooth, entire and yellow color after 4 days of incubation at 28°C on NA. In API 20NE, the reactions in reduction of D-glucose, and utilization of L-arabinose, D-mannose, D-mannitol, and N-acetyl-D-glucosamine are positive, but the reaction in nitrate reduction (NO3- → NO2-), reduction of nitrates to nitrogen, indole production, glucose acidification, arginine dihydrolase, urease, β-glucosidase (esculin hydrolysis), protease (gelatin hydrolysis), β-galactosidase (PNPG), D-maltose, gluconate, caprate, adipate, malate, citrate, and phenyl-acetate are negative. In API 32GN, the reaction in D-mannitol, and utilization of D-glucose, L-arabinose, L-histidine, 2-ketogluconate, 3-hydroxy-butyrate, L-proline, N-acetyl-D-glucosamine, acetate, lactate, L-alanine, and L-serine ware positive, but reaction in salicin, Dmelibiose, L-fucose, D-sorbitol, propionate, caprate, valerate, citrate, 4-hydroxy-benzoate, L-rhamnose, Dribose, inositol, D-sucrose, D-maltose, itaconate, suberate, malonate, 5-ketogluconate, glycogen, and 3-hydroxy- benzoate are negative. Positive reactions for enzymatic activity in alkaline phosphatase, esterase (C4), leucine arylamidase, acid phosphatase, naphtol-AS-BIphosphohydrolase, and α-glucosidase at API ZYM, but negative reactions for esterase lipase (C8), lipase (C14), valine arylamidase, cystine arylamidase, trypsin, α-chymotrypsin, α-galactosidase, β-galactosidase, β-glucuronidase, β-glucosidase, N-acetyl-β-glucosaminidase, α-mannosidase, and α-fucosidase. Strain NOY11-2 (=NIBRBAC000509782) was isolated from an agricultural soil sample, Gyeonggi-do, Korea (37°09ʹ07.3ʺN 126°58ʹ46.8ʺE).

    3.9. Description of Paenibacillus montanisoli ROW11-2

    Cells are Gram-staining-negative and rod-shape. Cell size is 4.1-4.8 μm. Colonies are circular, smooth, entire and white color after 4 days of incubation at 28°C on R2A. In API 20NE, the reactions in nitrate reduction (NO3- → NO2-), and utilization of L-arabinose, Dmannose, and phenyl-acetate are positive, but the reaction in indole production, glucose acidification, arginine dihydrolase, urease, β-glucosidase (esculin hydrolysis), protease (gelatin hydrolysis), β-galactosidase (PNPG), D-glucose, D-mannitol, N-acetyl-D-glucosamine, D-maltose, gluconate, caprate, adipate, malate, and citrate are positive. In API 32GN, the reaction in L-arabinose are positive, but reaction in D-mannitol, D-glucose, salicin, D-melibiose, L-fucose, D-sorbitol, propionate, caprate, valerate, citrate, L-histidine, 2-ketogluconate, 3-hydroxy-butyrate, 4-hydroxy-benzoate, L-proline, L-rhamnose, N-acetyl-D-glucosamine, Dribose, inositol, D-sucrose, D-maltose, itaconate, suberate, malonate, acetate, lactate, L-alanine, 5-ketogluconate, glycogen, 3-hydroxy-benzoate, and L-serine are negative. Enzymatic activity positive reaction was observed alkaline phosphatase, esterase (C4), esterase lipase (C8), leucine arylamidase, valine arylamidase, cystine arylamidase, α-chymotrypsin, acid phosphatase, and naphtol-AS-BI-phosphohydrolase at API ZYM, but negative reactions for lipase (C14), trypsin, α-galactosidase, β-galactosidase, β-glucuronidase, α-glucosidase, β-glucosidase, N-acetyl-β-glucosaminidase, α-mannosidase, and α-fucosidase. Strain ROW11-2 (=NIBR BAC000509783) was isolated from an agricultural soil sample, Gyeongsangnam-do, Korea (35°19ʹ59.8ʺN 128° 12ʹ34.7ʺE).

    3.10. Description of Paenibacillus frigoriresistens TW11

    Cells are Gram-staining-positive and rod-shape. Cell size is 3.8-4.5 μm. Colonies are circular, smooth, entire and yellow color after 4 days of incubation at 28°C on NA. In API 20NE, the reactions in nitrate reduction (NO3- → NO2-), and utilization of D-glucose, L-arabinose, D-mannose, D-mannitol, N-acetyl-D-glucosamine, D-maltose, and gluconate are positive, but the reaction in indole production, glucose acidification, arginine dihydrolase, urease, β-glucosidase (esculin hydrolysis), protease (gelatin hydrolysis), caprate, adipate, malate, citrate, and phenyl-acetate are negative. In API 32GN, the reaction in D-mannitol, and utilization of D-glucose, salicin, D-melibiose, L-arabinose, L-histidine, L-rhamnose, N-acetyl-D-glucosamine, D-ribose, D-sucrose, D-maltose, and glycogen are positive, but reaction in L-fucose, D-sorbitol, propionate, caprate, valerate, citrate, 2-ketogluconate, 3-hydroxy-butyrate, 4-hydroxy-benzoate, L-proline, inositol, itaconate, suberate, malonate, acetate, lactate, L-alanine, 5-ketogluconate, 3-hydroxy-benzoate, and L-serine are negative. Positive reactions for enzymatic activity in esterase (C4), esterase lipase (C8), leucine arylamidase, acid phosphatase, naphtol-AS-BI-phosphohydrolase, α-galactosidase, β-galactosidase, and α-glucosidase at API ZYM, but negative reactions for alkaline phosphatase, lipase (C14), valine arylamidase, cystine arylamidase, trypsin, α-chymotrypsin, β-glucuronidase, β-glucosidase, N-acetyl-β-glucosaminidase, α-mannosidase, and α- fucosidase. Strain TW11 (=NIBRBAC000509788) was isolated from an agricultural soil sample, Gyeonggi-do, Korea (37°08ʹ16.5ʺN 126°58ʹ12.9ʺE).

    ACKNOWLEDGEMENTS

    This work was supported by a grant from the National Institute of Biological Resources (NIBR), funded by the Ministry of Environment (MOE) of the Republic of Korea (NIBR202203112).

    CRediT authorship contribution statement

    OB Lim: Writing and Inverstigation. JS Lee: Inverstigation. H Lee: Conceptualization and Supervision. KE Lee: Data curation. IT Cha: Project administration. WJ Chi: Project administration. D-U Kim: Supervison, Funding acquisition, Resources, Writing-Reviewing and editing.

    Declaration of Competing Interest

    The authors declare no conflicts of interest.

    Figure

    KJEB-41-3-215_F1.gif

    Scanning electron microscopy and transmission electron microscopy of the cells of the unrecorded strains isolated in this study. The strains were cultured at their optimal growth conditions. 1, GW31; 2, R6; 3, JN1306; 4, SW56; 5, TA-E7; 6, SY19; 7, SY31; 8, NOY11-2; 9, ROW11-2; 10, TW11.

    KJEB-41-3-215_F2.gif

    A neighbor-joining phylogenetic tree based on 16S rRNA gene sequences shows the relationship between the strains isolated in this study and their relatives. Bootstrap values (>70%) are shown above nodes for the neighbor-joining methods. Bar: 0.020 substitutions per nucleotide position.

    Table

    The taxonomic affiliations of isolated strains belonging to the phylum Actinomycetota, Bacillota, and Pseudomonadota

    <sup>a</sup>NA, nutrient agar; R2A, Reasoner’2A agar; TSA, tryptic soy agar

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    13. Yoon SH , SM Ha, SJ Kwon, JG Lim, YS Kim, HS Seo and JS Chun.2017. Introducing EzBioCloud: a taxonomically united database of 16S rRNA gene sequences and whole-genome assemblies. Int. J. Syst. Evol. Microbiol. 67:1613-1617.

    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