Nakamurella Leprariae Sp. Nov., Isolated From a Lichen Sample

Nakamurella Leprariae Sp. 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Nov., Isolated From a Lichen Sample De-Feng An, Shao-Juan Yang, Long-Qian Jiang, Xin-Yu Wang, Xiao-Yu Huang, and 8 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-484462/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 15 Dec, 2021 Read the published version in Archives of Microbiology → Version 1 posted 5 You are reading this latest preprint version Abstract A novel actinobacterium, YIM 132084 T , was isolated from Lepraria sp. lichen collected from Yunnan province, south-west PR China and identified by a polyphasic taxonomic approach. The strain was Gram-stain-positive, aerobic, catalase-positive, oxidase-negative, non-motile and coccus-shaped. Colonies were round, convex, smooth and light orange yellow in colour. It grew at 10–40 ℃ (optimum 28 ℃), at pH 6.0–11.0 (optimum pH 7.0) and in the presence of 0–4 % NaCl (optimum 0 %). Strain YIM 132084 T comprised diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylinositol as the major polar lipids, MK-8(H 4 ) as the predominant menaquinone, and anteiso -C 15:0 , anteiso -C 17:0 , iso -C 15:0 and iso -C 16:0 as major fatty acids. Strain YIM 132084 T had meso -diaminopimelic acid as the diagnostic diamino acid in the cell-wall peptidoglycan, and mannose, ribose, glucose and rhamnose as whole-cell sugars. The 16S rRNA gene sequence showed high level of similarity with Nakamurella flavida KCTC 19127 T (97. 7%) and Nakamurella flava CGMCC 4.7524 T (97.7%). The G + C content of the genomic DNA was 72.4 mol%. Strain YIM 132084 T showed an average nucleotide identity value of 76.1 % and 74.9 %, a digital DNA-DNA hybridizations value of 20.9 % and 20.6 % with the reference strains Nakamurella flavida and Nakamurella flava based on draft genome sequences, respectively. The results of the phenotypic, chemotaxonomic and phylogenetic analyses, showed that strain YIM 132084 T represents a novel species of the genus Nakamurella , for which the name Nakamurella leprariae sp. nov. is proposed. The type strain is YIM 132084 T (= CGMCC 4.7667 T = NBRC 114280 T = KCTC 49367 T ). General Microbiology Applied & Industrial Microbiology Nakamurella Nakamurella leprariae sp. nov. Polyhasic taxonomy Lichen Figures Figure 1 Introduction The genus Nakamurella was proposed by Tao in 2004, as a substitute for the illegitimate genus Microsphaera by Yoshimi et al., at the same time, the family of Nakamurellaceae replaced Microsphaeraceae (Tao et al. 2004 and Yoshimi et al. 1996 ). Nakamurella species are distributed in different natural ecosystems, including activated sludge (Yoshimi et al. 1996 ), rock (Lee et al. 2008 ), soil (Yoon et al. 2007 ), faeces (Kim et al. 2017 ), lichen (Jiang et al. 2020 ), automobile air conditioning system (Chaudhary et al. 2021 ), plant (Yan et al. 2020 ) and bark (Tuo et al. 2016 ). At the time of writing, the genus Nakamurella is composed of 10 species with validly published names and two species with not validly published nomenclature ( https://www.bacterio.net/genus/nakamurella ) , and Nakamurella multipartita is the type species of this genus. During an investigate the diversity of cultivable actinobacteria from lichen samples collected in Yunnan province, south-west PR China, a new actinobacterium strain YIM 132084 T was isolated from Lepraria sp..The strain was identified by polyphasic approach, which indicated that it represented a new species of the genus Nakamurella . Material And Methods Isolation and Culture of Strains Lichens provide an extremely rich reservoir for the isolation of novel species. Strain YIM 132084 T was isolated from the lichen of genus Lepraria , The lichen Lepraria sp. sample was collected from Yunnan province (99°39′E, 22°23′N), south-west PR China. The lichen sample was transferred into a sterile paper bag and air-dried at 28 °C for 1 week, then washed three times with sterile water and homogenized with 18 ml of sterile 0.1% Na 4 P 2 O 7 using a sterile glass homogenizer. Strain YIM 132084 T was isolated using a standard dilution plate method on humic acid-vitamin (HV) agar (Hayakawa et al. 1987). The isolated colony was selected and further purified on YIM 38 medium (Li et al. 2016). Strain YIM 132084 T was stored in tubes of aqueous glycerine (20 %, v/v) and then in a -80 ℃ refrigerator. The reference strain, Nakamurella flavida KCTC 19127 T was gained from Korean Collection for Type Cultures (KCTC), Japan. Nakamurella flava CGMCC 4.7524 T was gained from China General Microbiological Culture Collection Centre (CGMCC). Phenotypic and Biochemical Tests Cultural characteristics of strain YIM 132084 T were observed after 3 days of incubation under aerobic conditions at 28 ℃ on YIM 38 medium. Morphological characteristics were observed by transmission electron microscopy (JEM-2100; JEOL). Growth in different culture media was performed using YIM 38 medium, tryptic soy agar (TSA, BD Difco), R2A agar (MB cell, Republic of Korea), Luria-Bertani (LB) agar, International Streptomyces Project Medium 2 (ISP 2, BD Difco), ISP 4 (BD Difco) at 28 °C for 3 days. Growth at different temperatures (4, 10, 15, 20, 25, 28, 30, 35, 37, 40 and 45℃) was tested on YIM 38 medium. The pH range for growth (pH 4.0-13.0, at intervals of 1.0 pH unit) was tested on YIM 38 at 28 °C. NaCl tolerance test for growth was performed using YIM 38 medium supplemented with different concentrations of NaCl (0-10 %, w/v, in increments of 1.0 %) at 28 °C. Anaerobic growth was tested after incubation on YIM 38 agar at 28 ℃ for 14 days using a GasPak EZ Anaerobe Pouch System (Becton Dickinson). Cell motility was determined in semisolid medium (Tittsler et al. 1936). Oxidase activity was determined by using 1 % (w/v) tetramethyl-p-phenylenediamine reagent and catalase activity was determined as the production of bubbles after the addition of 3 % (v/v) H 2 O 2 (Jiang et al. 2019). The Gram reaction of strain YIM 132084 T was examined using a standard Gram reaction and was confirmed by the 3 % KOH lysis test (Cerny 1978 and Buck 1982). Hydrolysis of starch, cellulose, tyrosine and casein, Tweens (20, 40, 60 and 80), gelatin liquefaction, H 2 S production, coagulation and peptonizationn of milk were tested using the methods described by Smibert et al. (1994). Susceptibility to antibiotics was tested on YIM 38 medium plate using filter paper containing the following antibiotics: ofloxacin (5 μg) , vancomycin (30 μg), ciprofloxacin (5 μg), norfloxacin (10 μg), polymyxin B (300 IU), gentamicin (10 μg), ampicillin (10 μg), chloramphenicol (30 μg), ceftriaxone (30 μg), penicillin G (10 IU), neomycin (30 μg), kanamycin (30 μg), streptomycin (50 μg ), novobiocin (5 μg), lincomycin (15 μg), tetracycline (30 μg). Sole carbon and nitrogen source utilization were determined using Biolog GEN III MicroPlate, other biochemical properties and enzyme activities were tested using API 20NE, API 50CH and API ZYM kits (bioMérieux) according to the manufacturer’s instructions. Phylogenetic Analysis and 16S rRNA Gene Sequencing Extraction of genomic DNA and PCR amplification of the 16S rRNA gene were done as described by Li et al. (2007). The purified product was cloned by using the pEASY-T1 sample cloning kit to obtain the almost-complete 16S rRNA gene sequence. The sequence obtained was compared with available 16S rRNA gene sequences of validly named species using the EzBioCloud server databases (https://www.ezbiocloud.net/) (Yoon et al. 2017). Phylogenetic trees were constructed with neighbour-joining (Saitou et al. 1987), maximum-likelihood algorithms (Tamura et al. 2011) and maximum parsimony (Fitch 1971) methods using the software package MEGA version 7.0 (Kumar et al. 2016). Kimura’s two-parameter model was used to calculate evolutionary distance matrices (Kimura 1980). Bootstrap values were calculated based on 1000 replications (Felsenstein 1985). Genomic Analysis The draft genome sequence of strain YIM 132084 T and Nakamurella flavida KCTC 19127 T were determined using the Illumina NovaSeq PE150 sequencing platform. The processed reads data were assembled using SOAPdenovo version 2.04 short sequence group assembly software (Li et al. 2008). The average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values were determined based on the genome sequences of YIM 132084 T and closely related species of Nakamurella using the EzBioCloud server databases and formula 2 Genome-to-Genome Distance Calculator website ( http://ggdc.dsmz.de/ggdc.php ) (Meier-Kolthoff et al. 2013), respectively. Gene annotations were conducted through the NCBI prokaryotic genome annotation pipeline. Chemotaxonomic Analysis The strain YIM 132084 T and the reference strains were cultured on YIM 38 agar at 28 °C for 3 days to obtain the amount needed for chemotaxonomic characterization. Polar lipids were extracted and analyzed by the method of Minnikin et al. (1984). Menaquinones were extracted by the method of Collins et al. (1977) and detected by HPLC (Tamaoka et al. 1983). The composition of cellular fatty acids were extracted and analyzed according to the standard protocol of the Microbial Identifcation System (MIDI) (Sasser 1990 and Kämpfer et al. 1996). Cell wall amino acids and whole cell sugars were extracted, detected and analyzed according to procedures described by Schleifer and Kandler (1972) and Tang et al. (2009). Results And Discussion Phenotypic and Biochemical Tests Cells of strain YIM 132084 T were Gram-stain-positive, aerobic, non-spore forming, non-motile, coccus-shaped and 0.7-0.9 µm in a diameter (Fig. S1). Colonies were round, convex, smooth and light orange yellow on YIM 38 agar at 28 °C for 3 days. The strain was found to growth on ISP 2, R2A, TSA, LB and YIM 38 agar, No growth occurs on ISP 4 agar. The growth range of strain YIM 132084 T at 10-40 ℃ (optimum 28 ℃), at pH 6.0-11.0 (optimum pH 7.0) and at 0-4 % NaCl (optimum 0 %). Hydrolysis of starch, cellulose, tyrosine and casein, Tweens (40, 60 and 80), gelatin liquefaction, H 2 S production, coagulation and peptonisation of milk were negative, except for hydrolysis of Tween 20. Susceptibility to ofloxacin, vancomycin, ciprofloxacin, norfloxacin, polymyxin B, gentamicin, chloramphenicol, neomycin, kanamycin were positive, susceptibility to ampicillin, ceftriaxone, penicillin G, streptomycin, novobiocin, lincomycin and tetracycline were negative. In the API ZYM tests, alkaline phosphatase, esterase (C 4 ), esterase lipase (C 8 ), leucine arylamidase, valine arylamidase, cystine arylamidase, acid phosphatase, naphthol-AS-BI-phosphohydrolase and α -glucosidase activities were positive, but lipase (C 14 ), β -glucuronidase, N -acetyl- β -glucosaminidase and α-fucosidase activities were negative. In the API 20NE strips, hydrolysis of L-arginine, urease, esculine and PNPG were positive, but other tests were negative. In the API 50CH strips, acid was produced from D-glucose, D-fructose, D-mannose, esculin citrate, D-maltose, D-sucrose, D-mycose, D-turanose and D-lyxose. The detailed physiological and biochemical characteristics of strain YIM 132084 T are shown in the species description and Table 1. Phylogenetic Analysis and 16S rRNA Gene Sequencing The almost-complete 16S rRNA gene sequence of strain YIM 132084 T was 1480bp (GenBank accession number MZ050064). Phylogenetic analyses based on the 16S rRNA gene sequence of strain YIM 132084 T indicated that it should be recognized as a member of the genus Nakamurella . Strain YIM 132084 T showed a high level of similarity with Nakamurella flavida KCTC 19127 T (97. 7 %) and Nakamurella flava CGMCC 4.7524 T (97.7 %). Phylogenetic trees were constructed by the neighbour-joining, maximum-likelihood algorithms and maximum parsimony based on the 16S rRNA gene sequence (Fig. 1, Fig. S2 and Fig. S3). The results of three tree-making algorithms showed that strain YIM 132084 T groups within the genus Nakamurella . Genomic Analysis Based on the draft genome sequencing, strain YIM 132084 T contained 39 contigs, with a total length of 4,472,446 bp and an N50 length of 232,774 bp (GenBank accession number JAERWK000000000). Based on the genomic annotation, the genome of strain YIM 132084 T contains 4,101 genes, included 4,009 protein-coding genes, 3 rRNA genes, 46 tRNA genes, 3 ncRNA genes and 40 pseudogenes. The DNA G+C content of strain YIM 132084 T was determined to be 72.4 mol% based on the draft genome. The ANI value between strain YIM 132084 T and the type strains of Nakamurella flavida KCTC 19127 T and Nakamurella flava CGMCC 4.7524 T were 76.1 and 74.9 %, respectively. The ANI value was lower than the 95.0% cut-off for species demarcation (Richter et al. 2009). The dDDH value between strain YIM 132084 T and the type strain: Nakamurella flavida KCTC 19127 T and Nakamurella flava CGMCC 4.7524 T were 20.9 and 20.6 %, respectively, which were much lower than the threshold value (70 %) recommended for distinguishing novel prokaryotic species. Chemotaxonomic Analysis The polar lipids profile of strain YIM 132084 T contained the predominant compounds diphosphatidylglycerol (DPG), phosphatidylethanolamine (PE), phosphatidylinositol (PI), an unidentified aminophospholipid (APL), an unidentified glycolipid (GL) and two unidentified phosphoglycolipids (PGL1-2) (Fig. S4). The predominant menaquinone was MK-8(H 4 ) in agreement with the genus Nakamurella (Chaudhary et al. 2021), in addition, MK-8(H 2 ) and MK-7(H 4 ) were detected in strain YIM 132084 T . The major cellular fatty acids consist of anteiso -C 15:0 (27.9 %), anteiso -C 17:0 (20.7 %), iso -C 15:0 (12.5 %) and is o-C 16:0 (16.0 %), which were similar to other members of the genus Nakamurella . The fatty acids composition and content comparison between strain YIM 132084 T and other closely related species of the genus Nakamurella are shown in Table 2. Strain YIM 132084 T had meso -diaminopimelic acid as the diagnostic diamino acid in the cell-wall peptidoglycan, which concurs with the members of the genus Nakamurella . The whole-cell sugars detected in strain YIM 132084 T were mannose, ribose, glucose and rhamnose. In conclusion, based on phenotypic, chemotaxonomic and phylogenetic analyses, strain YIM 132084 T is considered to represent a novel species of genus Nakamurella , for which the name Nakamurella leprariae sp. nov. is proposed. Description of Nakamurella leprariae sp. nov. Nakamurella leprariae (le.pra’ri.ae. N.L. gen. n. leprariae referring to the isolation of the organism from the lichen genus Lepraria ). Cells are Gram-stain-positive, catalase-positive, oxidase-negative, aerobic, non-motile, non-spore-forming and coccus-shaped (0.7-0.9 µm in diameter). Colonies on YIM 38 medium are round, smooth and convex, light orange yellow in colour. Growth occurs at 10-40 ℃ (optimum 28 ℃), at pH 6.0-11.0 (optimum pH 7.0) and at 0-4 % NaCl (optimum 0 %). Hydrolysis of starch, cellulose, tyrosine and casein, Tweens (40, 60 and 80), gelatin liquefaction, H 2 S production, coagulation and peptonisation of milk are negative, except for hydrolysis of Tween 20. In the Biolog GEN III system, the following substrates are used as a source of energy: β -methyl-D-glucoside, N -acetyl-D-glucosamine, N -acetyl-β-D-mannosamine, N -acetyl-D-galactosamine, D-mannose, D-fructose, D-galactose, D-mannitol, D-arabitol, myo-inositol, glycerol, D-glucose-6-phosphate, D-fructose-6-phosphate, D-aspartic acid, L-aspartic acid, L-glutamic acid, L-histidine, L-pyroglutamic acid, L-serine, D-glucuronic acid, D-saccharic acid, L-lactic acid, citric acid, α -keto-glutaric acid, D-malic acid, L-malic acid, bromo-succinic acid. The major polar lipids are diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylinositol. The predominant menaquinone is MK-8(H 4 ). The major fatty acids are anteiso -C 15:0 , anteiso -C 17:0 , iso -C 15:0 and iso -C 16:0 . Strain YIM 132084 T contain meso -diaminopimelic acid as the diagnostic diamino acid in the cell-wall peptidoglycan, and mannose, ribose, glucose and rhamnose as whole-cell sugars. The G+C content of the genomic DNA is 72.4 mol%. The type strain, YIM 132084 T (=CGMCC 4.7667 T = NBRC 114280 T =KCTC 49367 T ) was isolated from Lepraria sp. lichen collected from Yunnan province, south-west PR China. The GenBank accession number for the 16S rRNA gene sequence and draft genome sequence of strain YIM 132084 T are MZ050064 and JAERWK000000000, respectively. Abbreviations ANI, Average Nucleotide Identity; dDDH, digital DNA-DNA hybridization; DPG, diphosphatidylglycerol; PE, phosphatidylethanolamine; PI, phosphatidylinositol; APL, aminophospholipid; PGL, phosphoglycolipids; GL, glycolipid. Declarations Acknowledgements This research was funded by National Natural Science Foundation of China (32060001) and Major research project of Guangxi for science and technology (AA18242026). Author contributions De-Feng An and Shao-Juan Yang performed the experiments and wrote the manuscript; Long-Qian Jiang collected the lichen samples; Xiao-Yu Huang, Xue-Mei Chen, Ming-Qun Fan, Gui-Ding Li, and Lei Lang analyzed the data; Xin-Yun Wang identified the lichen samples; Yi Jiang and Ming-Guo Jiang guided the experiments and revised the manuscript; Cheng-Lin Jiang and Li-Song Wang designed the study. Conflict of interest The authors declare that they have no conflicts of interest. References Buck JD (1982) Nonstaining (KOH) method for determinationof gram reactions of marine bacteria. Appl Environ Microbiol 44:992–993 Cerny G (1978) Studies on the aminopeptidase test for the distinction of Gram-negative from Gram-positive bacteria. Applied Microbiology and Biotechnology 5(2):113-122 Chaudhary DK, Lee H, Dahal RH, Kim DY, Cha IT, Lee KE, Kim DU (2021) Nakamurella aerolata sp. Nov., Isolated from an Automobile Air Conditioning System. 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Int J Syst Bacteriol 46:519–525 Tables Table 1 Differential characteristics between strain YIM 132084 T and closely related species of the genus Nakamurella Characteristic 1 2 3 Isolation source Lichen Soil Mentha haplocalyx Briq. Colony colour Light orange yellow Light-yellow Brilliant orange yellow Cell size (µm) 0.7–0.9 0.6–1.2 a 1.0–1.8 b Growth at (°C) 10–40 4–35 4–40 pH 6–11 5–9 6–10 NaCl concentration (%, w/v) 0–4 0–3 0–5 Acidification of D-glucose - - + Hydrolysis of: Tween 20 + - + Tween 40 - - + Starch - + + Casein - - + Gelatin - + + Assimilation of: D-Glucose - - + L-Arabinose - - + D-Mannose - - + D-Mannitol - - + N -acetyl-D-glucosamine - - + Potassium gluconate - - + Enzyme activity: Cystine arylamidase + + - Trypsin - + - Chymotrypsin - + - β -galactosidase - + + β -glucosidase - + + Acid produced from: Glycol - - + Erythritol - - + L-arabinose - - + D-ribose - + - L-xylose - + + L-sorbose - + - N -acetylglucosamine - + + D-cellobiose - - + D-maltose + - + D-melibiose - - + D-trehalose + - + Inulin - + - D-melezitose - - + D-raffinose - + + D-gentiobiose - - + D-turanose + - + Susceptibility to Antibiotics: Norfloxacin + + - Polymyxin B + + - Neomycin + + - Kanamycin + + - Streptomycin - + - Novobiocin - + - Lincomycin - + - Tetracycline - + - DNA G + C content (mol%) 72.4 72.4 71.6 b Strains: 1, YIM 132084 T ; 2, Nakamurella flavida KCTC 19127 T ; 3, Nakamurella flava CGMCC 4.7524 T . +, Positive; −, negative. All data were obtained from this study except where indicated. Milk coagulation and peptonization, H 2 S production, hydrolysis of cellulose, tyrosine , Tween 60 and Tween 80 were negative in both strains. In API 20NE tests, all strains were positive for hydrolysis of L-arginine, urease, esculine and PNPG. In the API ZYM kits, all strains were positive for alkaline phosphatase, esterase (C 4 ), esterase lipase (C 8 ), leucine arylamidase, valine arylamidase, acid phosphatase, naphthol-AS-BI-phosphohydrolase, α -glucosidase. In the API 50CH kits, all strains were positive for acid production from D-glucose, D-fructose, D-mannose, esculin citrate, D-sucrose and D-lyxose. a Data from Yoon et al. (2007 ) b Data from Yan et al. (2020) Table 2. Cellular fatty acid compositions of strain YIM 132084 T and other closely related species of the genus Nakamurella Fatty acid 1 2 3 Straight-chain C 16:0 5.8 14.5 6.9 C 17:0 1.2 10.5 1.7 C 18:0 3.0 3.4 4.3 Branched anteiso-C 15:0 27.9 37.2 21.7 anteiso-C 16:0 0.4 2.1 1.4 anteiso-C 17:0 20.7 10.5 13.8 iso-C 14:0 0.7 0.5 2.3 iso-C 15:0 12.5 13.0 12.4 iso-C 16:0 16.0 3.8 7.1 iso-C 17:0 8.2 1.8 7.2 Summed Feature 3 * 1.5 0.9 5.4 Strains: 1, YIM 132084 T ; 2, Nakamurella flavida KCTC 19127 T ; 3, Nakamurella flava CGMCC 4.7524 T Values are percentages of total fatty acids. The major fatty acids (greater than 10.0 %) are shown bold. The data of YIM 132084 T , Nakamurella flavida KCTC 19127 T and Nakamurella flava CGMCC 4.7524 T were obtained from this study. * Summed features represent groups of two fatty acids that could not be separated by HPLC with the Microbial Identification System (MIDI, Inc.). Summed feature 3 consisted of C 16:1 ω 6 c and/or C 16:1 ω 7 c . Supplementary Files 132084supplementarymaterials.docx Cite Share Download PDF Status: Published Journal Publication published 15 Dec, 2021 Read the published version in Archives of Microbiology → Version 1 posted Editorial decision: Major revisions 08 Jun, 2021 Reviews received at journal 08 May, 2021 Reviewers invited by journal 08 May, 2021 Editor assigned by journal 07 May, 2021 First submitted to journal 30 Apr, 2021 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-484462","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":25895692,"identity":"9c7989b2-85a2-4a2d-b4ed-df782a7af33d","order_by":0,"name":"De-Feng An","email":"","orcid":"","institution":"Yunnan Institute of Microbiology","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"De-Feng","middleName":"","lastName":"An","suffix":""},{"id":25895693,"identity":"b32a5af5-cf66-4c05-855b-2ffd18d58870","order_by":1,"name":"Shao-Juan Yang","email":"","orcid":"","institution":"Yunnan Institute of Microbiology","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Shao-Juan","middleName":"","lastName":"Yang","suffix":""},{"id":25895694,"identity":"cb902903-e522-4e96-a151-d158562b3f9f","order_by":2,"name":"Long-Qian Jiang","email":"","orcid":"","institution":"Yunnan Institute of Microbiology","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Long-Qian","middleName":"","lastName":"Jiang","suffix":""},{"id":25895695,"identity":"73784498-ed20-4267-bdfe-f1b858b68407","order_by":3,"name":"Xin-Yu Wang","email":"","orcid":"","institution":"Kunming Institute of Botany Chinese Academy of Sciences","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Xin-Yu","middleName":"","lastName":"Wang","suffix":""},{"id":25895696,"identity":"ff6e36ff-2b97-462e-92b8-1d2f1e065de2","order_by":4,"name":"Xiao-Yu Huang","email":"","orcid":"","institution":"Yunnan Institute of Microbiology","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Xiao-Yu","middleName":"","lastName":"Huang","suffix":""},{"id":25895697,"identity":"53b3b371-e864-4d33-8131-a7e6d6c80e9a","order_by":5,"name":"Lei Lang","email":"","orcid":"","institution":"Yunnan Institute of Microbiology","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Lei","middleName":"","lastName":"Lang","suffix":""},{"id":25895698,"identity":"b09b2c4d-9a81-40a2-8a72-86fea2d3d9cc","order_by":6,"name":"Xue-Mei Chen","email":"","orcid":"","institution":"Yunnan Institute of Microbiology","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Xue-Mei","middleName":"","lastName":"Chen","suffix":""},{"id":25895699,"identity":"9e3d5317-fbfb-4702-baa5-6728f70dbc87","order_by":7,"name":"Ming-Qun Fan","email":"","orcid":"","institution":"Yunnan Institute of Microbiology","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Ming-Qun","middleName":"","lastName":"Fan","suffix":""},{"id":25895700,"identity":"b2057b73-6348-441c-9068-56f8ed303915","order_by":8,"name":"Gui-Ding Li","email":"","orcid":"","institution":"Yunnan Institute of Microbiology","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Gui-Ding","middleName":"","lastName":"Li","suffix":""},{"id":25895701,"identity":"51e2c89a-0215-4a67-a46f-255788bdf89e","order_by":9,"name":"Ming-Guo Jiang","email":"","orcid":"","institution":"Guangxi University for Nationalities","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Ming-Guo","middleName":"","lastName":"Jiang","suffix":""},{"id":25895702,"identity":"ffb6303d-3565-4ff4-8b35-c9e8acd5684c","order_by":10,"name":"Li-Song Wang","email":"","orcid":"","institution":"Kunming Institute of Botany Chinese Academy of Sciences","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Li-Song","middleName":"","lastName":"Wang","suffix":""},{"id":25895703,"identity":"a096189c-5367-43a8-88b2-2655eb169108","order_by":11,"name":"Cheng-Lin Jiang","email":"","orcid":"","institution":"Yunnan Institute of Microbiology","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Cheng-Lin","middleName":"","lastName":"Jiang","suffix":""},{"id":25895704,"identity":"dfdef726-b104-4f89-8a11-4a001d032366","order_by":12,"name":"Yi Jiang","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAzklEQVRIiWNgGAWjYBACxmYwJSEH4bKRoMWYeC0wkNhAtBbmduZnD7+2WaRvuN1jwPCh7DAD/+wGQg5jMzeWbZPI3XDnjAHjjHOHGSTuHCCkhcFMWhKk5UaOATNv22EGA4kEQlrYv4G0pBuAtPwlTguPmeTHNokEsBZGIrWUSTOckzCceSOt4GDPuXQeiRsEtBj2H98m+aOsTp7vRvLGBz/KrOX4ZxDS0gAMaF5gdCgcYGAAIgYe/OqBQB7kuB9/gIwGgmpHwSgYBaNgpAIAoctAZ92MWF0AAAAASUVORK5CYII=","orcid":"https://orcid.org/0000-0002-1847-252X","institution":"Yunnan Institute of Microbiology, Yunnan University","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Yi","middleName":"","lastName":"Jiang","suffix":""}],"badges":[],"createdAt":"2021-05-01 16:35:17","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-484462/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-484462/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s00203-021-02626-7","type":"published","date":"2021-12-15T08:49:36+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":9048327,"identity":"0b7851d4-afa2-46cc-9f07-d4567287e896","added_by":"auto","created_at":"2021-05-11 14:16:40","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":80907,"visible":true,"origin":"","legend":"Neighbour-joining phylogenetic tree based on 16S rRNA gene sequences, showing the position of strain YIM 132084T in relation to its nearest phylogenetical neighbours. Numbers at nodes indicate the level of bootstrap support (\u003e 50 %) based on 1000 resamplings. Klenkia marina YIM M13156T (LT746188) was used as an outgroup. Bar, 0.005 substitutions per nucleotide position.","description":"","filename":"fig1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-484462/v1/e265eb9a891cb73283ad421b.jpg"},{"id":16468734,"identity":"6486f334-3a03-40d1-a545-eb215d67a6fe","added_by":"auto","created_at":"2021-12-15 08:49:43","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":402840,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-484462/v1/ccd75042-cc99-440e-9c44-3d39546d9886.pdf"},{"id":9048693,"identity":"2b88fc47-8a84-401b-8737-66c84f1bb6a3","added_by":"auto","created_at":"2021-05-11 14:19:41","extension":"docx","order_by":6,"title":"","display":"","copyAsset":false,"role":"supplement","size":476791,"visible":true,"origin":"","legend":"","description":"","filename":"132084supplementarymaterials.docx","url":"https://assets-eu.researchsquare.com/files/rs-484462/v1/cd200ab3a053f5ae4a52abc8.docx"}],"financialInterests":"","formattedTitle":"\u003cp\u003e\u003cem\u003eNakamurella Leprariae\u003c/em\u003e Sp. Nov., Isolated From a Lichen Sample\u003c/p\u003e","fulltext":[{"header":"Introduction","content":" \u003cp\u003eThe genus \u003cem\u003eNakamurella\u003c/em\u003e was proposed by Tao in 2004, as a substitute for the illegitimate genus \u003cem\u003eMicrosphaera\u003c/em\u003e by Yoshimi et al., at the same time, the family of \u003cem\u003eNakamurellaceae\u003c/em\u003e replaced \u003cem\u003eMicrosphaeraceae\u003c/em\u003e (Tao et al. \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2004\u003c/span\u003e and Yoshimi et al. \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e1996\u003c/span\u003e). \u003cem\u003eNakamurella\u003c/em\u003e species are distributed in different natural ecosystems, including activated sludge (Yoshimi et al. \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e1996\u003c/span\u003e), rock (Lee et al. \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2008\u003c/span\u003e), soil (Yoon et al. \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2007\u003c/span\u003e), faeces (Kim et al. \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2017\u003c/span\u003e), lichen (Jiang et al. \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2020\u003c/span\u003e), automobile air conditioning system (Chaudhary et al. \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2021\u003c/span\u003e), plant (Yan et al. \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) and bark (Tuo et al. \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). At the time of writing, the genus \u003cem\u003eNakamurella\u003c/em\u003e is composed of 10 species with validly published names and two species with not validly published nomenclature (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.bacterio.net/genus/nakamurella\u003c/span\u003e\u003c/span\u003e\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003e)\u003c/span\u003e, and \u003cem\u003eNakamurella multipartita\u003c/em\u003e is the type species of this genus. During an investigate the diversity of cultivable actinobacteria from lichen samples collected in Yunnan province, south-west PR China, a new actinobacterium strain YIM 132084\u003csup\u003eT\u003c/sup\u003e was isolated from \u003cem\u003eLepraria\u003c/em\u003e sp..The strain was identified by polyphasic approach, which indicated that it represented a new species of the genus \u003cem\u003eNakamurella\u003c/em\u003e.\u003c/p\u003e "},{"header":"Material And Methods","content":"\u003cp\u003e\u003cstrong\u003eIsolation and Culture of Strains\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eLichens provide an extremely rich reservoir for the isolation of novel species. Strain YIM 132084\u003csup\u003eT\u003c/sup\u003e was isolated from the lichen of genus \u003cem\u003eLepraria\u003c/em\u003e, The lichen \u003cem\u003eLepraria \u003c/em\u003esp. sample was collected from Yunnan province (99\u0026deg;39\u0026prime;E, 22\u0026deg;23\u0026prime;N), south-west PR China. The lichen sample was transferred into a sterile paper bag and air-dried at 28 \u0026deg;C for 1 week, then washed three times with sterile water and homogenized with 18 ml of sterile 0.1% Na\u003csub\u003e4\u003c/sub\u003eP\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e7\u003c/sub\u003e using a sterile glass homogenizer. Strain YIM 132084\u003csup\u003eT\u003c/sup\u003e was isolated using a standard dilution plate method on humic acid-vitamin (HV) agar (Hayakawa et al. 1987). The isolated colony was selected and further purified on YIM 38 medium (Li et al. 2016). Strain YIM 132084\u003csup\u003eT\u003c/sup\u003e was stored in tubes of aqueous glycerine (20 %, v/v) and then in a -80 ℃ refrigerator. The reference strain, \u003cem\u003eNakamurella\u0026nbsp;flavida \u003c/em\u003eKCTC 19127\u003csup\u003eT\u003c/sup\u003e was gained from Korean Collection for Type Cultures (KCTC), Japan. \u003cem\u003eNakamurella\u0026nbsp;flava \u003c/em\u003eCGMCC 4.7524\u003csup\u003eT\u003c/sup\u003e was gained from China General Microbiological Culture Collection Centre (CGMCC).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePhenotypic and Biochemical Tests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eCultural characteristics of strain YIM 132084\u003csup\u003eT\u003c/sup\u003e were observed after 3 days of incubation under aerobic conditions at 28 ℃ on YIM 38 medium. Morphological characteristics were observed by transmission electron microscopy (JEM-2100; JEOL). Growth in different culture media was performed using YIM 38 medium, tryptic soy agar (TSA, BD Difco), R2A agar (MB cell, Republic of Korea), Luria-Bertani (LB) agar, International Streptomyces Project Medium 2 (ISP 2, BD Difco), ISP 4 (BD Difco) at 28 \u0026deg;C for 3 days. Growth at different temperatures (4, 10, 15, 20, 25, 28, 30, 35, 37, 40 and 45℃) was tested on YIM 38 medium. The pH range for growth (pH 4.0-13.0, at intervals of 1.0 pH unit) was tested on YIM 38 at 28 \u0026deg;C. NaCl tolerance test for growth was performed using YIM 38 medium supplemented with different concentrations of NaCl (0-10 %, w/v, in increments of 1.0 %) at 28 \u0026deg;C. Anaerobic growth was tested after incubation on YIM 38 agar at 28 ℃ for 14 days using a GasPak EZ Anaerobe Pouch System (Becton Dickinson). Cell motility was determined in semisolid medium (Tittsler et al. 1936). Oxidase activity was determined by using 1 % (w/v) tetramethyl-p-phenylenediamine reagent and catalase activity was determined as the production of bubbles after the addition of 3 % (v/v) H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2 \u003c/sub\u003e(Jiang et al. 2019). The Gram reaction of strain YIM 132084\u003csup\u003eT\u003c/sup\u003e was examined using a standard Gram reaction and was confirmed by the 3 % KOH lysis test (Cerny 1978 and Buck 1982). Hydrolysis of starch, cellulose, tyrosine and casein, Tweens (20, 40, 60 and 80), gelatin liquefaction, H\u003csub\u003e2\u003c/sub\u003eS production, coagulation and peptonizationn of milk were tested using the methods described by Smibert et\u0026nbsp;al. (1994). Susceptibility to antibiotics was tested on YIM 38 medium plate using filter paper containing the following antibiotics: ofloxacin (5 \u0026mu;g) , vancomycin (30 \u0026mu;g), ciprofloxacin (5 \u0026mu;g), norfloxacin (10 \u0026mu;g), polymyxin B (300 IU), gentamicin (10 \u0026mu;g), ampicillin (10 \u0026mu;g), chloramphenicol (30 \u0026mu;g), ceftriaxone (30 \u0026mu;g), penicillin G (10 IU), neomycin (30 \u0026mu;g), kanamycin (30 \u0026mu;g), streptomycin (50 \u0026mu;g ), novobiocin (5 \u0026mu;g), lincomycin (15 \u0026mu;g), tetracycline (30 \u0026mu;g). Sole carbon and nitrogen source utilization were determined using Biolog GEN III MicroPlate, other biochemical properties and enzyme activities were tested using API 20NE, API 50CH and API ZYM kits (bioMérieux) according to the manufacturer\u0026rsquo;s instructions.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePhylogenetic Analysis and 16S rRNA Gene Sequencing\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eExtraction of genomic DNA and PCR amplification of the 16S rRNA gene were done as described by Li et al. (2007). The purified product was cloned by using the pEASY-T1 sample cloning kit to obtain the almost-complete 16S rRNA gene sequence. The sequence obtained was compared with available 16S rRNA gene sequences of validly named species using the EzBioCloud server databases (https://www.ezbiocloud.net/) (Yoon et al. 2017). Phylogenetic trees were constructed with neighbour-joining (Saitou et al. 1987), maximum-likelihood algorithms (Tamura et al. 2011) and maximum parsimony (Fitch 1971) methods using the software package MEGA version 7.0 (Kumar et al. 2016). Kimura\u0026rsquo;s two-parameter model was used to calculate evolutionary distance matrices (Kimura 1980). Bootstrap values were calculated based on 1000 replications (Felsenstein 1985).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eGenomic Analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe draft genome sequence of strain YIM 132084\u003csup\u003eT\u003c/sup\u003e and \u003cem\u003eNakamurella\u0026nbsp;flavida \u003c/em\u003eKCTC 19127\u003csup\u003eT\u003c/sup\u003e were determined using the\u0026nbsp;Illumina NovaSeq PE150 sequencing platform. The processed reads data were assembled using SOAPdenovo version 2.04 short sequence group assembly software (Li et al. 2008). The average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values were determined based on the genome sequences of YIM 132084\u003csup\u003eT\u003c/sup\u003e and closely related species of \u003cem\u003eNakamurella\u003c/em\u003e using the EzBioCloud server databases and formula 2 Genome-to-Genome Distance Calculator website (\u003ca href=\"http://ggdc.dsmz.de/ggdc.php\"\u003ehttp://ggdc.dsmz.de/ggdc.php\u003c/a\u003e) (Meier-Kolthoff et al. 2013), respectively. Gene annotations were conducted through the NCBI prokaryotic genome annotation pipeline.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eChemotaxonomic Analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe strain YIM 132084\u003csup\u003eT\u003c/sup\u003e and the reference strains were cultured on YIM 38 agar at 28 \u0026deg;C for 3 days to obtain the amount needed for chemotaxonomic characterization. Polar lipids were extracted and analyzed by the method of Minnikin et al. (1984). Menaquinones were extracted by the method of Collins et al. (1977) and detected by HPLC (Tamaoka et al. 1983). The composition of cellular fatty acids were extracted\u0026nbsp;and analyzed according to the standard protocol of the Microbial Identifcation System (MIDI) (Sasser 1990 and K\u0026auml;mpfer et al. 1996). Cell wall amino acids and whole cell sugars were extracted, detected and analyzed according to procedures described by Schleifer and Kandler (1972) and Tang et al. (2009).\u003c/p\u003e"},{"header":"Results And Discussion","content":"\u003cp\u003e\u003cstrong\u003ePhenotypic and Biochemical Tests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eCells of strain YIM 132084\u003csup\u003eT\u003c/sup\u003e were Gram-stain-positive, aerobic, non-spore forming, non-motile, coccus-shaped and 0.7-0.9 \u0026micro;m in a diameter (Fig. S1). Colonies were round, convex, smooth and light orange yellow on YIM 38 agar at 28 \u0026deg;C for 3 days. The strain was found to growth on ISP 2, R2A, TSA, LB and YIM 38 agar, No growth occurs on ISP 4 agar. The growth range of strain YIM 132084\u003csup\u003eT\u003c/sup\u003e at 10-40 ℃ (optimum 28 ℃), at pH 6.0-11.0 (optimum pH 7.0) and at 0-4 % NaCl (optimum 0 %). Hydrolysis of starch, cellulose, tyrosine and casein, Tweens (40, 60 and 80), gelatin liquefaction, H\u003csub\u003e2\u003c/sub\u003eS production, coagulation and peptonisation of milk were negative, except for hydrolysis of Tween 20. Susceptibility to ofloxacin, vancomycin, ciprofloxacin, norfloxacin, polymyxin B, gentamicin, chloramphenicol, neomycin, kanamycin were positive, susceptibility to ampicillin, ceftriaxone, penicillin G, streptomycin, novobiocin, lincomycin and tetracycline were negative. In the API ZYM tests, alkaline phosphatase, esterase (C\u003csub\u003e4\u003c/sub\u003e), esterase lipase (C\u003csub\u003e8\u003c/sub\u003e), leucine arylamidase, valine arylamidase, cystine arylamidase, acid phosphatase, naphthol-AS-BI-phosphohydrolase and \u003cem\u003e\u0026alpha;\u003c/em\u003e-glucosidase activities were positive, but lipase (C\u003csub\u003e14\u003c/sub\u003e), \u003cem\u003e\u0026beta;\u003c/em\u003e-glucuronidase, \u003cem\u003eN\u003c/em\u003e-acetyl-\u003cem\u003e\u0026beta;\u003c/em\u003e-glucosaminidase and \u0026alpha;-fucosidase activities were negative. In the API 20NE strips, hydrolysis of L-arginine, urease, esculine and PNPG were positive, but other tests were negative. In the API 50CH strips, acid was produced from D-glucose, D-fructose, D-mannose, esculin citrate, D-maltose, D-sucrose, D-mycose, D-turanose and D-lyxose. The detailed physiological and biochemical characteristics of strain YIM 132084\u003csup\u003eT\u003c/sup\u003e are shown in the species description and Table 1.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePhylogenetic Analysis and 16S rRNA Gene Sequencing\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe almost-complete 16S rRNA gene sequence of strain YIM 132084\u003csup\u003eT \u003c/sup\u003ewas 1480bp (GenBank accession number MZ050064). Phylogenetic analyses based on the 16S rRNA gene sequence of strain YIM 132084\u003csup\u003eT\u003c/sup\u003e indicated that it should be recognized as a member of the genus \u003cem\u003eNakamurella\u003c/em\u003e. Strain YIM 132084\u003csup\u003eT \u003c/sup\u003eshowed a high level of similarity with \u003cem\u003eNakamurella\u0026nbsp;flavida\u003c/em\u003e KCTC 19127\u003csup\u003eT \u003c/sup\u003e(97. 7 %) and \u003cem\u003eNakamurella\u0026nbsp;flava \u003c/em\u003eCGMCC 4.7524\u003csup\u003eT \u003c/sup\u003e(97.7 %). Phylogenetic trees were constructed by the neighbour-joining, maximum-likelihood algorithms and maximum parsimony based on the 16S rRNA gene sequence (Fig. 1, Fig. S2 and Fig. S3). The results of three tree-making algorithms showed that strain YIM 132084\u003csup\u003eT\u003c/sup\u003e groups within the genus \u003cem\u003eNakamurella\u003c/em\u003e.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eGenomic Analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eBased on the draft genome sequencing, strain YIM 132084\u003csup\u003eT \u003c/sup\u003econtained 39 contigs, with a total length of 4,472,446 bp and an N50 length of 232,774 bp (GenBank accession number JAERWK000000000). Based on the genomic annotation, the genome of strain YIM 132084\u003csup\u003eT\u003c/sup\u003e contains 4,101 genes, included 4,009 protein-coding genes, 3 rRNA genes, 46 tRNA genes, 3 ncRNA genes and 40 pseudogenes. The DNA G+C content of strain YIM 132084\u003csup\u003eT\u003c/sup\u003e was determined to be 72.4 mol% based on the draft genome. The ANI value between strain YIM 132084\u003csup\u003eT\u003c/sup\u003e and the type strains of \u003cem\u003eNakamurella\u0026nbsp;\u003c/em\u003e\u003cem\u003eflavida \u003c/em\u003eKCTC 19127\u003csup\u003eT \u003c/sup\u003eand \u003cem\u003eNakamurella\u0026nbsp;flava \u003c/em\u003eCGMCC 4.7524\u003csup\u003eT \u003c/sup\u003ewere 76.1 and 74.9 %, respectively. The ANI value was lower than the 95.0% cut-off for species demarcation (Richter et al. 2009). The dDDH value between strain YIM 132084\u003csup\u003eT\u003c/sup\u003e and the type strain: \u003cem\u003eNakamurella\u003c/em\u003e\u003cem\u003e\u0026nbsp;flavida \u003c/em\u003eKCTC 19127\u003csup\u003eT \u003c/sup\u003eand \u003cem\u003eNakamurella\u0026nbsp;flava \u003c/em\u003eCGMCC 4.7524\u003csup\u003eT \u003c/sup\u003ewere 20.9 and 20.6 %, respectively, which were much lower than the threshold value (70 %) recommended for distinguishing novel prokaryotic species.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eChemotaxonomic Analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe polar lipids profile of strain YIM 132084\u003csup\u003eT\u003c/sup\u003e contained the predominant compounds diphosphatidylglycerol (DPG), phosphatidylethanolamine (PE), phosphatidylinositol (PI), an unidentified aminophospholipid (APL), an unidentified glycolipid (GL) and two unidentified phosphoglycolipids (PGL1-2) (Fig. S4). The predominant menaquinone was MK-8(H\u003csub\u003e4\u003c/sub\u003e) in agreement with the genus \u003cem\u003eNakamurella\u003c/em\u003e (Chaudhary et al. 2021), in addition, MK-8(H\u003csub\u003e2\u003c/sub\u003e) and MK-7(H\u003csub\u003e4\u003c/sub\u003e) were detected in strain YIM 132084\u003csup\u003eT\u003c/sup\u003e. The major cellular fatty acids consist of \u003cem\u003eanteiso\u003c/em\u003e-C\u003csub\u003e15:0\u003c/sub\u003e (27.9 %), \u003cem\u003eanteiso\u003c/em\u003e-C\u003csub\u003e17:0 \u003c/sub\u003e(20.7 %), \u003cem\u003eiso\u003c/em\u003e-C\u003csub\u003e15:0 \u003c/sub\u003e(12.5 %) and \u003cem\u003eis\u003c/em\u003eo-C\u003csub\u003e16:0\u003c/sub\u003e (16.0 %), which were similar to other members of the genus \u003cem\u003eNakamurella\u003c/em\u003e. The fatty acids composition and content comparison between strain YIM 132084\u003csup\u003eT\u003c/sup\u003e and other closely related species of the genus \u003cem\u003eNakamurella\u003c/em\u003e are shown in Table 2. Strain YIM 132084\u003csup\u003eT\u003c/sup\u003e had \u003cem\u003emeso\u003c/em\u003e-diaminopimelic acid as the diagnostic diamino acid in the cell-wall peptidoglycan, which concurs with the members of the genus \u003cem\u003eNakamurella\u003c/em\u003e. The whole-cell sugars detected in strain YIM 132084\u003csup\u003eT\u003c/sup\u003e were mannose, ribose, glucose and rhamnose.\u003c/p\u003e\n\u003cp\u003eIn conclusion, based on phenotypic, chemotaxonomic and phylogenetic analyses, strain YIM 132084\u003csup\u003eT\u003c/sup\u003e is considered to represent a novel species of genus \u003cem\u003eNakamurella\u003c/em\u003e, for which the name \u003cem\u003eNakamurella \u003c/em\u003e\u003cem\u003eleprariae \u003c/em\u003esp. nov. is proposed.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDescription of \u003cem\u003eNakamurella leprariae\u003c/em\u003esp. nov.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eNakamurella leprariae\u003c/em\u003e (le.pra\u0026rsquo;ri.ae. N.L. gen. n. \u003cem\u003eleprariae\u003c/em\u003e referring to the isolation of the organism from the lichen genus \u003cem\u003eLepraria\u003c/em\u003e).\u003c/p\u003e\n\u003cp\u003eCells are Gram-stain-positive, catalase-positive, oxidase-negative, aerobic, non-motile, non-spore-forming and coccus-shaped (0.7-0.9 \u0026micro;m in diameter). Colonies on YIM 38 medium are round, smooth and convex, light orange yellow in colour. Growth occurs at 10-40 ℃ (optimum 28 ℃), at pH 6.0-11.0 (optimum pH 7.0) and at 0-4 % NaCl (optimum 0 %). Hydrolysis of starch, cellulose, tyrosine and casein, Tweens (40, 60 and 80), gelatin liquefaction, H\u003csub\u003e2\u003c/sub\u003eS production, coagulation and peptonisation of milk are negative, except for hydrolysis of Tween 20. In the Biolog GEN III system, the following substrates are used as a source of energy: \u003cem\u003e\u0026beta;\u003c/em\u003e-methyl-D-glucoside, \u003cem\u003eN\u003c/em\u003e-acetyl-D-glucosamine, \u003cem\u003eN\u003c/em\u003e-acetyl-\u0026beta;-D-mannosamine, \u003cem\u003eN\u003c/em\u003e-acetyl-D-galactosamine, D-mannose, D-fructose, D-galactose, D-mannitol, D-arabitol, myo-inositol, glycerol, D-glucose-6-phosphate, D-fructose-6-phosphate, D-aspartic acid, L-aspartic acid, L-glutamic acid, L-histidine, L-pyroglutamic acid, L-serine, D-glucuronic acid, D-saccharic acid, L-lactic acid, citric acid, \u003cem\u003e\u0026alpha;\u003c/em\u003e-keto-glutaric acid, D-malic acid, L-malic acid, bromo-succinic acid. The major polar lipids are diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylinositol. The predominant menaquinone is MK-8(H\u003csub\u003e4\u003c/sub\u003e). The major fatty acids are \u003cem\u003eanteiso\u003c/em\u003e-C\u003csub\u003e15:0\u003c/sub\u003e, \u003cem\u003eanteiso\u003c/em\u003e-C\u003csub\u003e17:0\u003c/sub\u003e, \u003cem\u003eiso\u003c/em\u003e-C\u003csub\u003e15:0\u003c/sub\u003e and \u003cem\u003eiso\u003c/em\u003e-C\u003csub\u003e16:0\u003c/sub\u003e. Strain YIM 132084\u003csup\u003eT\u003c/sup\u003e contain \u003cem\u003emeso\u003c/em\u003e-diaminopimelic acid as the diagnostic diamino acid in the cell-wall peptidoglycan, and mannose, ribose, glucose and rhamnose as whole-cell sugars. The G+C content of the genomic DNA is 72.4 mol%. The type strain, YIM 132084\u003csup\u003eT\u003c/sup\u003e (=CGMCC 4.7667\u003csup\u003eT\u003c/sup\u003e = NBRC 114280\u003csup\u003eT\u003c/sup\u003e=KCTC 49367\u003csup\u003eT\u003c/sup\u003e) was isolated from \u003cem\u003eLepraria\u003c/em\u003e sp. lichen collected from Yunnan province, south-west PR China. The GenBank accession number for the 16S rRNA gene sequence and draft genome sequence of strain YIM 132084\u003csup\u003eT\u003c/sup\u003e are MZ050064 and JAERWK000000000, respectively.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cp\u003eANI, Average Nucleotide Identity; dDDH, digital DNA-DNA hybridization; DPG, diphosphatidylglycerol; PE, phosphatidylethanolamine; PI, phosphatidylinositol; APL, aminophospholipid; PGL, phosphoglycolipids; GL, glycolipid.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research was funded by National Natural Science Foundation of China (32060001) and Major research project of Guangxi for science and technology (AA18242026).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eDe-Feng An and Shao-Juan Yang performed the experiments and wrote the manuscript; Long-Qian Jiang collected the lichen samples; Xiao-Yu Huang, Xue-Mei Chen, Ming-Qun Fan, Gui-Ding Li, and Lei Lang analyzed the data; Xin-Yun Wang identified the lichen samples; Yi Jiang and Ming-Guo Jiang guided the experiments and revised the manuscript; Cheng-Lin Jiang and Li-Song Wang designed the study.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of interest \u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no conflicts of interest.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eBuck JD (1982) Nonstaining (KOH) method for determinationof gram reactions of marine bacteria. 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Int J Syst Bacteriol 46:519\u0026ndash;525\u003c/li\u003e\n\u003c/ol\u003e"},{"header":"Tables","content":"\u003cdiv class=\"gridtable\"\u003e\n\u003ctable id=\"Tab1\" border=\"1\"\u003e\u003ccaption\u003e\n\u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\n\u003cdiv class=\"CaptionContent\"\u003e\n\u003cp\u003eDifferential characteristics between strain YIM 132084\u003csup\u003eT\u003c/sup\u003e and closely related species of the genus \u003cem\u003eNakamurella\u003c/em\u003e\u003c/p\u003e\n\u003c/div\u003e\n\u003c/caption\u003e\n\u003cthead\u003e\n\u003ctr\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003eCharacteristic\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e1\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/th\u003e\n\u003cth align=\"left\"\u003e\n\u003cp\u003e3\u003c/p\u003e\n\u003c/th\u003e\n\u003c/tr\u003e\n\u003c/thead\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eIsolation source\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eLichen\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eSoil\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u003cem\u003eMentha haplocalyx\u003c/em\u003e Briq.\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eColony colour\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eLight orange yellow\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eLight-yellow\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eBrilliant orange yellow\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eCell size (\u0026micro;m)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.7\u0026ndash;0.9\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0.6\u0026ndash;1.2\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e1.0\u0026ndash;1.8\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eGrowth at (\u0026deg;C)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e10\u0026ndash;40\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4\u0026ndash;35\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e4\u0026ndash;40\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003epH\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e6\u0026ndash;11\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e5\u0026ndash;9\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e6\u0026ndash;10\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eNaCl concentration (%, w/v)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0\u0026ndash;4\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0\u0026ndash;3\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e0\u0026ndash;5\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eAcidification of D-glucose\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eHydrolysis of:\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTween 20\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTween 40\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eStarch\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eCasein\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eGelatin\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eAssimilation of:\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eD-Glucose\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eL-Arabinose\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eD-Mannose\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eD-Mannitol\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u003cem\u003eN\u003c/em\u003e-acetyl-D-glucosamine\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003ePotassium gluconate\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eEnzyme activity:\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eCystine arylamidase\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTrypsin\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eChymotrypsin\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u003cem\u003e\u0026beta;\u003c/em\u003e-galactosidase\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u003cem\u003e\u0026beta;\u003c/em\u003e-glucosidase\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eAcid produced from:\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eGlycol\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eErythritol\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eL-arabinose\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eD-ribose\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eL-xylose\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eL-sorbose\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e\u003cem\u003eN\u003c/em\u003e-acetylglucosamine\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eD-cellobiose\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eD-maltose\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eD-melibiose\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eD-trehalose\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eInulin\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eD-melezitose\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eD-raffinose\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eD-gentiobiose\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eD-turanose\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eSusceptibility to\u003c/p\u003e\n\u003cp\u003eAntibiotics:\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eNorfloxacin\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003ePolymyxin B\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eNeomycin\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eKanamycin\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eStreptomycin\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eNovobiocin\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eLincomycin\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eTetracycline\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e+\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e-\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003eDNA G\u0026thinsp;+\u0026thinsp;C content (mol%)\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e72.4\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e72.4\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd align=\"left\"\u003e\n\u003cp\u003e71.6\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eStrains: 1, YIM 132084\u003csup\u003eT\u003c/sup\u003e; 2, \u003cem\u003eNakamurella\u0026nbsp;flavida \u003c/em\u003eKCTC 19127\u003csup\u003eT\u003c/sup\u003e; 3, \u003cem\u003eNakamurella\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eflava \u003c/em\u003eCGMCC 4.7524\u003csup\u003eT\u003c/sup\u003e. +, Positive; \u0026minus;, negative. All data were obtained from this study except where indicated.\u003c/p\u003e\n\u003cp\u003eMilk coagulation and peptonization, H\u003csub\u003e2\u003c/sub\u003eS production, hydrolysis of cellulose, \u0026nbsp;tyrosine , Tween 60 and Tween 80 were negative in both strains. In API 20NE tests, all strains were positive for hydrolysis of L-arginine, urease, esculine and PNPG. In the API ZYM kits, all strains were positive for alkaline phosphatase, esterase (C\u003csub\u003e4\u003c/sub\u003e), esterase lipase (C\u003csub\u003e8\u003c/sub\u003e), leucine arylamidase, valine arylamidase, acid phosphatase, naphthol-AS-BI-phosphohydrolase, \u003cem\u003e\u0026alpha;\u003c/em\u003e-glucosidase. In the API 50CH kits, all strains were positive for acid production from D-glucose, D-fructose, D-mannose, esculin citrate, D-sucrose and D-lyxose.\u003c/p\u003e\n\u003cp\u003e\u003csup\u003ea\u003c/sup\u003eData from Yoon et al. (2007 )\u003c/p\u003e\n\u003cp\u003e\u003csup\u003eb\u003c/sup\u003eData from Yan et al. (2020)\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTable 2. \u003c/strong\u003eCellular fatty acid compositions of strain YIM 132084\u003csup\u003eT\u003c/sup\u003e and other closely related species of the genus \u003cem\u003eNakamurella\u003c/em\u003e\u003c/p\u003e\n\u003ctable border=\"1\"\u003e\n\u003ctbody\u003e\n\u003ctr\u003e\n\u003ctd width=\"148\"\u003e\n\u003cp\u003eFatty acid\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"136\"\u003e\n\u003cp\u003e1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"142\"\u003e\n\u003cp\u003e2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"142\"\u003e\n\u003cp\u003e3\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"148\"\u003e\n\u003cp\u003eStraight-chain\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"136\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"142\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"142\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"148\"\u003e\n\u003cp\u003eC\u003csub\u003e16:0\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"136\"\u003e\n\u003cp\u003e5.8\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"142\"\u003e\n\u003cp\u003e\u003cstrong\u003e14.5\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"142\"\u003e\n\u003cp\u003e6.9\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"148\"\u003e\n\u003cp\u003eC\u003csub\u003e17:0\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"136\"\u003e\n\u003cp\u003e1.2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"142\"\u003e\n\u003cp\u003e\u003cstrong\u003e10.5\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"142\"\u003e\n\u003cp\u003e1.7\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"148\"\u003e\n\u003cp\u003eC\u003csub\u003e18:0\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"136\"\u003e\n\u003cp\u003e3.0\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"142\"\u003e\n\u003cp\u003e3.4\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"142\"\u003e\n\u003cp\u003e4.3\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"148\"\u003e\n\u003cp\u003eBranched\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"136\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"142\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"142\"\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"148\"\u003e\n\u003cp\u003eanteiso-C\u003csub\u003e15:0\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"136\"\u003e\n\u003cp\u003e\u003cstrong\u003e27.9\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"142\"\u003e\n\u003cp\u003e\u003cstrong\u003e37.2\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"142\"\u003e\n\u003cp\u003e\u003cstrong\u003e21.7\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"148\"\u003e\n\u003cp\u003eanteiso-C\u003csub\u003e16:0\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"136\"\u003e\n\u003cp\u003e0.4\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"142\"\u003e\n\u003cp\u003e2.1\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"142\"\u003e\n\u003cp\u003e1.4\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"148\"\u003e\n\u003cp\u003eanteiso-C\u003csub\u003e17:0\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"136\"\u003e\n\u003cp\u003e\u003cstrong\u003e20.7\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"142\"\u003e\n\u003cp\u003e\u003cstrong\u003e10.5\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"142\"\u003e\n\u003cp\u003e\u003cstrong\u003e13.8\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"148\"\u003e\n\u003cp\u003eiso-C\u003csub\u003e14:0\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"136\"\u003e\n\u003cp\u003e0.7\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"142\"\u003e\n\u003cp\u003e0.5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"142\"\u003e\n\u003cp\u003e2.3\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"148\"\u003e\n\u003cp\u003eiso-C\u003csub\u003e15:0\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"136\"\u003e\n\u003cp\u003e\u003cstrong\u003e12.5\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"142\"\u003e\n\u003cp\u003e\u003cstrong\u003e13.0\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"142\"\u003e\n\u003cp\u003e\u003cstrong\u003e12.4\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"148\"\u003e\n\u003cp\u003eiso-C\u003csub\u003e16:0\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"136\"\u003e\n\u003cp\u003e\u003cstrong\u003e16.0\u003c/strong\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"142\"\u003e\n\u003cp\u003e3.8\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"142\"\u003e\n\u003cp\u003e7.1\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"148\"\u003e\n\u003cp\u003eiso-C\u003csub\u003e17:0\u003c/sub\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"136\"\u003e\n\u003cp\u003e8.2\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"142\"\u003e\n\u003cp\u003e1.8\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"142\"\u003e\n\u003cp\u003e7.2\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003ctr\u003e\n\u003ctd width=\"148\"\u003e\n\u003cp\u003eSummed Feature 3\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"136\"\u003e\n\u003cp\u003e1.5\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"142\"\u003e\n\u003cp\u003e0.9\u003c/p\u003e\n\u003c/td\u003e\n\u003ctd width=\"142\"\u003e\n\u003cp\u003e5.4\u003c/p\u003e\n\u003c/td\u003e\n\u003c/tr\u003e\n\u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003eStrains: 1, YIM 132084\u003csup\u003eT\u003c/sup\u003e; 2, \u003cem\u003eNakamurella\u003c/em\u003e\u003cem\u003e\u0026nbsp;flavida \u003c/em\u003eKCTC 19127\u003csup\u003eT\u003c/sup\u003e; 3, \u003cem\u003eNakamurella\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eflava \u003c/em\u003eCGMCC 4.7524\u003csup\u003eT \u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003eValues are percentages of total fatty acids. The major fatty acids (greater than 10.0 %) are shown bold. The data of YIM 132084\u003csup\u003eT\u003c/sup\u003e, \u003cem\u003eNakamurella\u0026nbsp;flavida \u003c/em\u003eKCTC 19127\u003csup\u003eT \u003c/sup\u003eand \u003cem\u003eNakamurella flava \u003c/em\u003eCGMCC 4.7524\u003csup\u003eT \u003c/sup\u003ewere obtained from this study.\u003c/p\u003e\n\u003cp\u003e\u003csup\u003e*\u003c/sup\u003eSummed features represent groups of two fatty acids that could not be separated by HPLC with the Microbial Identification System (MIDI, Inc.). Summed feature 3 consisted of C\u003csub\u003e16:1\u003c/sub\u003e \u003cem\u003e\u0026omega;\u003c/em\u003e6\u003cem\u003ec\u003c/em\u003e and/or C\u003csub\u003e16:1 \u003c/sub\u003e\u003cem\u003e\u0026omega;\u003c/em\u003e7\u003cem\u003ec\u003c/em\u003e.\u003c/p\u003e\n\u003c/div\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":true,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"archives-of-microbiology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"aomi","sideBox":"Learn more about [Archives of Microbiology](https://www.springer.com/journal/203)","snPcode":"203","submissionUrl":"https://submission.nature.com/new-submission/203/3","title":"Archives of Microbiology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Nakamurella, Nakamurella leprariae sp. nov., Polyhasic taxonomy, Lichen","lastPublishedDoi":"10.21203/rs.3.rs-484462/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-484462/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eA novel actinobacterium, YIM 132084\u003csup\u003eT\u003c/sup\u003e, was isolated from \u003cem\u003eLepraria\u003c/em\u003e sp. lichen collected from Yunnan province, south-west PR China and identified by a polyphasic taxonomic approach. The strain was Gram-stain-positive, aerobic, catalase-positive, oxidase-negative, non-motile and coccus-shaped. Colonies were round, convex, smooth and light orange yellow in colour. It grew at 10\u0026ndash;40 ℃ (optimum 28 ℃), at pH 6.0\u0026ndash;11.0 (optimum pH 7.0) and in the presence of 0\u0026ndash;4 % NaCl (optimum 0 %). Strain YIM 132084\u003csup\u003eT\u003c/sup\u003e comprised diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylinositol as the major polar lipids, MK-8(H\u003csub\u003e4\u003c/sub\u003e) as the predominant menaquinone, and \u003cem\u003eanteiso\u003c/em\u003e-C\u003csub\u003e15:0\u003c/sub\u003e, \u003cem\u003eanteiso\u003c/em\u003e-C\u003csub\u003e17:0\u003c/sub\u003e, \u003cem\u003eiso\u003c/em\u003e-C\u003csub\u003e15:0\u003c/sub\u003e and \u003cem\u003eiso\u003c/em\u003e-C\u003csub\u003e16:0\u003c/sub\u003e as major fatty acids. Strain YIM 132084\u003csup\u003eT\u003c/sup\u003e had \u003cem\u003emeso\u003c/em\u003e-diaminopimelic acid as the diagnostic diamino acid in the cell-wall peptidoglycan, and mannose, ribose, glucose and rhamnose as whole-cell sugars. The 16S rRNA gene sequence showed high level of similarity with \u003cem\u003eNakamurella flavida\u003c/em\u003e KCTC 19127\u003csup\u003eT\u003c/sup\u003e (97. 7%) and \u003cem\u003eNakamurella flava\u003c/em\u003e CGMCC 4.7524\u003csup\u003eT\u003c/sup\u003e (97.7%). The G\u0026thinsp;+\u0026thinsp;C content of the genomic DNA was 72.4 mol%. Strain YIM 132084\u003csup\u003eT\u003c/sup\u003e showed an average nucleotide identity value of 76.1 % and 74.9 %, a digital DNA-DNA hybridizations value of 20.9 % and 20.6 % with the reference strains \u003cem\u003eNakamurella flavida\u003c/em\u003e and \u003cem\u003eNakamurella flava\u003c/em\u003e based on draft genome sequences, respectively. The results of the phenotypic, chemotaxonomic and phylogenetic analyses, showed that strain YIM 132084\u003csup\u003eT\u003c/sup\u003e represents a novel species of the genus \u003cem\u003eNakamurella\u003c/em\u003e, for which the name \u003cem\u003eNakamurella leprariae\u003c/em\u003e sp. nov. is proposed. The type strain is YIM 132084\u003csup\u003eT\u003c/sup\u003e (=\u0026thinsp;CGMCC 4.7667\u003csup\u003eT\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;NBRC 114280\u003csup\u003eT\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;KCTC 49367\u003csup\u003eT\u003c/sup\u003e).\u003c/p\u003e","manuscriptTitle":"Nakamurella Leprariae Sp. Nov., Isolated From a Lichen Sample","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2021-05-11 14:16:39","doi":"10.21203/rs.3.rs-484462/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Major revisions","date":"2021-06-08T07:51:03+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2021-05-09T00:57:00+00:00","index":0,"fulltext":""},{"type":"reviewersInvited","content":"","date":"2021-05-08T04:37:00+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2021-05-08T02:39:00+00:00","index":"","fulltext":""},{"type":"submitted","content":"Archives of Microbiology","date":"2021-04-30T10:22:03+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"archives-of-microbiology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"aomi","sideBox":"Learn more about [Archives of Microbiology](https://www.springer.com/journal/203)","snPcode":"203","submissionUrl":"https://submission.nature.com/new-submission/203/3","title":"Archives of Microbiology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"74696ee8-a582-4fe0-8245-f3e6e537053d","owner":[],"postedDate":"May 11th, 2021","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":4231878,"name":"General Microbiology"},{"id":4231879,"name":"Applied \u0026 Industrial Microbiology"}],"tags":[],"updatedAt":"2021-12-15T08:49:36+00:00","versionOfRecord":{"articleIdentity":"rs-484462","link":"https://doi.org/10.1007/s00203-021-02626-7","journal":{"identity":"archives-of-microbiology","isVorOnly":false,"title":"Archives of Microbiology"},"publishedOn":"2021-12-15 08:49:36","publishedOnDateReadable":"December 15th, 2021"},"versionCreatedAt":"2021-05-11 14:16:39","video":"","vorDoi":"10.1007/s00203-021-02626-7","vorDoiUrl":"https://doi.org/10.1007/s00203-021-02626-7","workflowStages":[]},"version":"v1","identity":"rs-484462","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-484462","identity":"rs-484462","version":["v1"]},"buildId":"7rjqhiLT3MXkJMwkYKINL","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}