Jeotgalibacillus haloalkaliphilus sp. nov., a novel alkaliphilic and halotolerant bacterium, isolated from the estuary of the Fenhe River into the Yellow River

Jeotgalibacillus haloalkaliphilus sp. nov., a novel alkaliphilic and halotolerant bacterium, isolated from the estuary of the Fenhe River into the Yellow River | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Jeotgalibacillus haloalkaliphilus sp. nov., a novel alkaliphilic and halotolerant bacterium, isolated from the estuary of the Fenhe River into the Yellow River Ya-Fei Zhang, Yun-Lin Peng, Yong-Hui Xiao, Bing Yu, Ming-Yue Cao, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-3880322/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 01 May, 2024 Read the published version in Antonie van Leeuwenhoek → Version 1 posted 4 You are reading this latest preprint version Abstract A Gram-stain positive, aerobic, alkaliphilic and halotolerant bacterium, designated HH7-29 T , was isolated from the estuary of the Fenhe River into the Yellow River in Shanxi Province, PR China. Growth occurred at pH 6.0–12.0 (optimum, pH 8.0–8.5) and 15–40℃ (optimum, 32℃) with 0.5–24% NaCl (optimum, 2–9%). The predominant fatty acids (> 10.0%) were iso-C 15:0 and anteiso-C 15:0 . The major menaquinones were MK-7 and MK-8. The polar lipids were phosphatidylglycerol, diphosphatidylglycerol and two unidentified phospholipids. Phylogenetic analyses based on the 16S rRNA gene sequence revealed that strain HH7-29 T was a member of the genus Jeotgalibacillus , exhibiting high sequence similarity to the 16S rRNA gene sequences of Jeotgalibacillus alkaliphilus JC303 T (98.4%), Jeotgalibacillus salarius ASL-1 T (98.1%) and Jeotgalibacillus alimentarius YKJ-13 T (98.1%). The genomic DNA G + C content was 43.0%. Gene annotation showed that strain HH7-29 T had lower protein isoelectric points (pIs) and possessed genes related to ion transport and organic osmoprotectant uptake, implying its potential tolerance to salt and alkali. The average nucleotide identity, digital DNA–DNA hybridization values, amino acid identity values, and percentage of conserved proteins values between strain HH7-29 T and its related species were 71.1–83.8%, 19.5–27.4%, 66.5–88.4% and 59.8–76.6%, respectively. Based on the analyses of phenotypic, chemotaxonomic, phylogenetic and genomic features, strain HH7-29 T represents a novel species of the genus Jeotgalibacillus , for which the name Jeotgalibacillus haloalkaliphilus sp. nov. is proposed. The type strain is HH7-29 T (= KCTC 43417 T = MCCC 1K07541 T ). Jeotgalibacillus haloalkaliphilus polyphasic taxonomy estuary alkaliphilic and halotolerant Figures Figure 1 Figure 2 Figure 3 Figure 4 INTRODUCTION The genus Jeotgalibacillus , a member of the family Caryophanaceae within the order Caryophanales of the class Bacilli , was first identified by Yoon et al in 2001 with the description of Jeotgalibacillus alimentarius as the type species (Yoon et al. 2001 ). At the time of writing, the genus Jeotgalibacillus comprised 10 recognized species: Jeotgalibacillus alimentarius (Yoon et al. 2001 ), Jeotgalibacillus alkaliphilus (Srinivas et al. 2016 ), Jeotgalibacillus aurantiacus (Jiang et al. 2022 ), Jeotgalibacillus campisalis (Yoon et al. 2004 ), Jeotgalibacillus malaysiensis (Yaakop et al. 2015 ), Jeotgalibacillus marinus (Rüger and Richter 1979 ), Jeotgalibacillus proteolyticus (Li et al. 2018 ), Jeotgalibacillus salarius (Yoon et al. 2010 ), Jeotgalibacillus soli (Cunha et al. 2012 ) and Jeotgalibacillus terrae (Srinivas et al. 2016 ), among which Jeotgalibacillus campisalis and Jeotgalibacillus marinus were named through the reclassification of two Marinibacillus species (Yoon et al. 2010 ). Except for J. terrae and J. aurantiacus (from forest soil and wetland soil, respectively), the other species originated from saline or alkaline environments, such as marine saltern, salt pan, salted fermented food and alkaline sandy soil. These Jeotgalibacillus species represent a group of Gram-stain-positive, rod-shaped bacteria with DNA G + C content of 39.3–44.0%, containing MK-7 and MK-8 as the major quinones, and iso-C 15:0 and anteiso-C 15:0 as the major cellular fatty acids (Yaakop et al. 2015 ). Most of members exhibit a yellow pigmentation (Jiang et al. 2022 ), which may be associated with their self-defense against harsh environmental conditions (Soliev et al. 2011 ). Some species, such as J. malaysiensis and J. proteolyticus , have ability for secreting a multitude of enzymes (e.g., protease, gelatinase, esterase and β -glucosidase), which have important implications towards the biotechnological industry (Liew et al. 2018 ; Li et al. 2018 ). High salinity is one of the most challenging stresses for life on Earth. However, microorganisms have developed two mechanisms to maintain their cellular osmotic balance: by increasing the ion concentrations or by accumulating protective organic osmolytes in cells (Mongodin et al. 2005 ; Burg and Ferraris 2008 ). Jeotgalibacillus spp. were reported to be slightly halophilic, and some of these strains tolerate salt concentrations of up to 30% (w/v) (Yaakop et al. 2015 ). Earlier genomic analyses revealed that the genus Jeotgalibacillus features diverse genes resistant to salt, which help them improve their ecological fitness in alkaline saline ecosystems (Jiang et al. 2022 ). During our survey of the diversity of the culturable bacteria from the river confluences of Shanxi, China, we discovered a novel alkaliphilic and halotolerant bacterium belonging to the genus Jeotgalibacillus , designated as HH7-29 T . To better understand and use it, strain HH7-29 T was subjected to polyphasic taxonomic analyses involving phylogenetic, phenotypic and chemotaxonomic profiles. Furthermore, we also analyzed the genomic characteristics associated with alkaline saline tolerance to understand how it addressed the challenges of osmolality fluctuations. METHODS Strain isolation and culture conditions Fresh sediment samples were obtained from the estuary of the Fenhe River into the Yellow River (35°35′N, 110°47′E) of Shanxi, China. 1 g of thoroughly homogenized samples were tenfold serially diluted using sterile 1% NaCl, and spread onto 1/3 MA agar (MA; Difco). Single orange-yellow colony approximately 2 mm in diameter was picked and designated HH7-29 T . Single colony was transferred into TYS broth (0.5% tryptone, 0.1% yeast extract, 3.0% salt) for growth at 30℃ and stored at − 80℃ as glycerol suspensions (20%). For comparative study, the reference strain Jeotgalibacillus salarius KCTC 13257 T was obtained from the KCTC (Korean Collection for Type Cultures) and grown on TYS agar at 30°C. 16S rRNA gene sequencing and phylogenetic analysis For phylogenetic analysis of strain HH7-29 T , the almost-complete 16S rRNA gene sequence was amplified by PCR with primers 27F and 1492R (Weisburg et al. 1991 ). Following purification, the PCR products were ligated to the pMD19-T Vector (TaKaRa) and sequenced at Sangon Biotech (Shanghai, China) using an ABI 3730 DNA analyser. The sequence of strain HH7-29 T was compared to reference strains using EzBioCloud database. A multiple alignment of all the sequences was achieved by using CLUSTAL W algorithm (Thompson et al. 1994 ) integrated in MEGA 11 (Koichiro et al. 2021 ). Phylogenetic tree was reconstructed with MEAG 11 using the maximum-likelihood (ML) (Felsenstein 1981 ), neighbour-joining (NJ) (Saitou and Nei 1987 ), and minimum-evolution (ME) (Pardi et al. 2010 ) algorithm, and evaluated by bootstrap analysis with 1000 replications. Draft genome sequencing and genome sequences analysis Whole genome sequencing of strain HH7-29 T was conducted by Oebiotech (Shanghai, China) using the Illumina HiSeq 4000 system (Illumina, USA). The paired-end reads assembly were performed with ABySS v2.0 (Jackman et al. 2017 ). Genome annotation was performed with National Center for Biotechnology Information (NCBI) Prokaryotic Genome Annotation Pipeline (PGAP) (Tatusova et al. 2016) and the Rapid Annotations using Subsystems Technology (RAST) server (Aziz et al. 2008 ). The secondary metabolite biosynthesis gene clusters of genome sequences were annotated using antiSMASH bacterial version (Medema et al. 2011 ). The G + C content of the genomic DNA was determined based on the genome sequence. Genomes of closely related strains were retrieved from the NCBI GenBank database. The 815 single-copy orthologous clusters (OCs) (Jones et al. 1992 ) were selected using Proteinortho v5.16 (Lechner et al. 2011 ) and aligned separately with MUSCLE v3.8.31 (Edgar 2004 ). Phylogenetic trees based on the whole genome were reconstructed using MEGA 11. Average nucleotide identity (ANI) values were calculated using EzBioCloud web service ( https://www.ezbiocloud.net/tools/ani ) (Yoon et al. 2017 ), and digital DNA–DNA hybridization (dDDH) values were determined using the Genome-to-Genome Distance Calculator ( http://ggdc.dsmz.de/ggdc.php ) (Meier-Kolthoff et al. 2013 ). Average amino acid identity (AAI) values were calculated using an online AAI calculator server ( http://enve-omics.ce.gatech.edu/aai/ ) (Luo et al. 2014), and the percentage of conserved proteins (POCP) values were estimated based on a previously described approach (Qin et al. 2014 ). Growth and morphological characteristics The morphological of strain HH7-29 T was investigated after cultivation on TYS at 30℃ for 12 h. Cell size, shape and flagella were assessed with transmission electron microscopy (Tecnai G2 F20, FEI). The Gram reaction was tested using a Gram-stain kit (Solarbio, China) according to the manufacturer’s instructions. Temperature sensitivity was determined in TYS broth over a range of 4–45℃ (4, 10, 15, 20, 25, 28, 32, 37, 40 and 45℃). Salt tolerance was tested by using a modified medium (0.5% tryptone, 0.1% yeast extract, 0.5% MgCl 2 , 0.05% CaCl 2 , 0.1% KCl and 0.0001% FeSO 4 ) with varying NaCl concentrations (0–4% NaCl, increased at intervals of 0.5%; 4–30% NaCl, increased at intervals of 1%). The pH range for growth was tested in TYS broth at pH 5.0 − 10.0 (increments of 0.5 pH unit) by adding 50 mmol/L buffers [MES (pH 5.0 − 6.0), MOPS (pH 6.5 − 7.0), Tris (pH 7.5 − 8.5) and CHES (pH 9.0 − 10.0)]. Catalase activity was tested by assessing the formation of bubbles upon the addition of 3% (w/v) H 2 O 2 (Jones 1981 ). Oxidase activity was performed with oxidase strips (Merck, Germany). Anaerobic respiration was determined in Hungate tubes filled with TYS broth supplemented with 0.1% (w/v) potassium nitrate, 0.05% (w/v) cysteine hydrochloride and 0.05% (w/v) sodium sulfide. Hydrolyses of casein (0.5%, w/v), starch (0.5%, w/v), milk (0.5%, w/v), gelatin (0.5%, w/v), cellulose (0.5%, w/v), sodium alginate (0.5%, w/v) and Tweens (20, 40, 60 and 80, 1%, v/v) were tested as previously described by Smibert and Krieg (Smibert and Krieg 1981 ). Antibiotic susceptibility testing was performed according to the antibiotic disc-difusion method using 16 common antibiotics: ampicillin (10 µg), cephalexin (30 µg), erythromycin (15 µg), roxithromycin (15 µg), tetracycline (30 µg), oxytetracycline (30 µg), streptomycin (10 µg), kanamycin (30 µg), sulfamethoxazole (25 µg), sulfafurazole (25 µg), norfloxacin (10 µg), levofloxacin (5 µg), vancomycin (30 µg), polymyxin B (300 IU), licomycin (2 µg), and chloramphenicol (30 µg). Other physiological and biochemical characteristics were determined with API 20 NE and API ZYM systems (bioMérieux, France) following the manufacture instructions. Chemotaxonomic analysis For the analyses of fatty acids, respiratory quinones, and polar lipids, strain HH7-29 T and the reference strain J. salarius KCTC 13257 T were obtained from cultures grown in TYS broth at 30℃ for 12 h (logarithmic phase of growth). The harvested cells were washed twice in sterile 1% NaCl and then freeze dried. Cellular fatty acids were extracted, methylated, and identified using gas chromatography (Agilent 6850), following the instructions of the Sherlock Microbial Identification System (MIDI, version 6.1). Peaks were automatically integrated, and fatty acid names and percentages were determined using the Microbial Identification System (MIS) standard software. The polar lipids were extracted according to Komagata and Suzuki ( 1987 ) and determined using a 2D thin-layer chromatography (TLC) procedure on Merck silica gel 60 F254 plates. The staining reagents phosphomolybdic acids was used for detecting the total lipids, ninhydrin for lipids containing free amino groups, and molybdenum blue for phospholipids (Collins and Jones 1980 ). Respiratory quinones were extracted from freeze-dried cells using a mixture of chloroform/methanol (2:1, v/v), and the extract was analyzed by an LC–MS system (a Dionex Ultimate 3000 HPLC device connected to a Bruker impact HD mass spectrometer). RESULTS AND DISCUSSION Phylogenetic analysis based on 16S rRNA gene sequences The nearly full-length 16S rRNA gene sequence of strain HH7-29 T (1513 bp) has been deposited as OR856029 in the GenBank/ database. On pairwise comparison with the 16S rRNA gene sequences available in the EzBioCloud database, strain HH7-29 T exhibited the highest sequence similarities to J. alkaliphilus JC303 T (98.4%), followed by J. salarius ASL-1 T (98.1%), J. alimentarius YKJ-13 T (98.1%) and other Jeotgalibacillus members (94.6–98.0%). These values were lower than the threshold (98.7%) for separating different species (Chun et al. 2018 ). In the ML, NJ and ME phylogenetic trees (Figs. 1 , S1 and S2), strain HH7-29 T fell within a sub-cluster of the genus Jeotgalibacillus , which consisted of five Jeotgalibacillus species ( J. alkaliphilus , J. salarius , J. malaysiensis , J. alimentarius and J. terrae ). Within this sub-cluster, strain HH7-29 T formed an internally independent branch among adjacent strains. The taxonomic identification and evolutionary tree based on the 16S rRNA gene showed that strain HH7-29 T represented a potential novel species. Genome features and comparison The assembled genome sequencing of strain HH7-29 T (JAXQNN000000000) yielded a genome of 3284775 bp in length, which was composed of 28 contigs with an N50 value of 985906 bp (Table S1 ). The G + C content was 43.0%, which was in the range of 39.7–43.7% for other species of Jeotgalibacillus . A total of 3337 genes were predicted in the genome, including 3265 protein-coding genes, 16 pseudo genes and 56 RNA genes (6 rRNAs, 5 ncRNAs and 45 tRNAs). 1323 (39.6%) of all predicted genes were assigned through the RAST database. The abundance of carbohydrates metabolism (15.3%), protein metabolism (13.0%) and DNA metabolism (4.7%) in strain HH7-29 T fell within the range of the closely related species (Table S2). While numbers of genes associated with respiration and potassium metabolism in strain HH7-29 T were higher than that in closely related species. Meanwhile, the genes encoding hydrogenase were unique to strain HH7-29 T . Additionally, genomic comparisons showed that all the nine strains shared 1352 core genes in common, accounting for 37.3–42.1% of the annotated genome repertoire respectively (Fig. 2 ). The antiSMASH results showed that strain HH7-29 T harbors two sets of gene clusters required for carotenoid biosynthesis and siderophore biosynthesis, respectively. Additionally, the genes encoding peptidases such as ATP-dependent protease La, aminopeptidase and carboxylate peptidase were detected in the genome. In line with this, strain HH7-29 T had milk and casein degradation capacity (Fig. S3). In the phylogenetic trees based on 815 single-copy orthologous clusters (Figs. 3 , S4 and S5), strain HH7-29 T clustered within the clade that accommodated the genus Jeotgalibacillus , in which it formed a cluster with J. malaysiensis D5 T and J. salarius ASL-1 T . The ANI values between strain HH7-29 T and related Jeotgalibacillus species were 71.1–83.8% (Table S3). Moreover, the dDDH values between strain HH7-29 T and related species ranged from 19.5 to 27.4% (Table S3). These values are significantly lower than the threshold values for prokaryotic species delineation, which are 95–96% for ANI and 70% for dDDH (Kim et al. 2014 ; Meier-Kolthoff et al. 2013 ). Furthermore, the AAI and POCP values between strain HH7-29 T and related species were 66.5–88.4% and 59.8–76.6% (Table S4), which were within the range indicating the delimitation boundaries of the genus (above 65–95% and 50%, respectively) (Luo et al. 2014; Qin et al. 2014 ). These results confirmed that HH7-29 T represents a novel species within the genus Jeotgalibacillus . Salinity tolerance Under high-salinity conditions, the growth of microorganisms is severely limited. Surprisingly, strain HH7-29 T could grow in NaCl concentrations up to 24%, showing high salinity tolerance ability (Table 1 ). It has been reported that proteins of halophilic organisms have more acidic residues and lower isoelectric points (pIs) to accommodate high ionic strength (Mongodin et al. 2005 ). In this study, through comparison of pIs from HH7-29 T , the closely related strains, hyperhalophilic bacterium Salinibacter ruber DSM 13855 T , non-halophilic bacterium Escherichia coli K-12 T , these Jeotgalibacillus members exhibited strongly unimodal distributions of pI values and higher ratios of acidic residues to basic residues, S. ruber too (Fig. 4 ). The average pI values of all proteins in HH7-29 T and the closely related strains was 6.5, significantly lower than protein pIs of E. coli K-12 T with 7.1 (Table S5). Through functional annotation of genes, strain HH7-29 T possesses betaine/carnitine/choline transporter, betaine/proline/choline transporter and glycine betaine transporter (Table S6). As osmoprotection, compatible solutes (e.g., glycine betaine, proline) accumulation could increase to maintain cellular osmotic balance in saline conditions (Mongodin et al. 2005 ; Burg and Ferraris 2008 ). Though the genes encoding choline dehydrogenases and betaine aldehyde dehydrogenase, which are responsible for synthesizing glycine betaine from choline (Vargas et al. 2008 ; Wargo 2013 ), were lacked, HH7-29 T could encode two glutamate synthases (UFB30_01585, UFB30_01590). As an organic osmolyte, glutamate could mediate a global transcription switch at genomic promoters that vary in their response to salt (Burg and Ferraris 2008 ). Besides, the genome contained 13 sodium/proton antiporters, five Trk system potassium uptake protein, three sodium/calcium antiporter and two potassium/proton antiporters (Table S6), which maintained high intracellular ion concentrations within cells to achieve osmotic equilibrium. Furthermore, those sodium or potassium/proton antiporters may catalyze proton accumulation coupled with sodium efflux, thereby maintaining a cytoplasmic pH well below the high external pH in strain HH7-29 T , supporting cytoplasmic pH homeostasis and alkali tolerance(Ito et al. 2004 ; Krulwich et al. 2009 ). Collectively, strain HH7-29 T could not only accumulate protective organic osmolytes but also increase intracellular ion concentrations to maintain its tolerance to salt, alkali and osmolarity fluctuations. Table 1 Differential characteristics of strain HH7-29 T and type strains of closely related species. Characteristic 1 2 3 4 5 6 7 8 9 Growth range of: Temperature (optimum, ℃) 15–40℃ (32℃) 4–40℃ (30℃) * 10–45℃ (30–35℃) 4–50℃ (37℃) 10–45℃ (30–35℃) 15–40℃ (37℃) 4–39℃ (30℃) 10–40℃ (33℃) 15–40℃ (30–37℃) NaCl (optimum, %) 0.5–24% (2–9%) 0–18% (2%) * 0–20% (3–12%) 0–30% (10%) 0–20% (2–5%) 0–9% (2%) 0–15% (2–3%) 0–10% (2%) 0–9% (0–1%) pH (optimum) 6.0–12.0 (8.0–8.5) 6.0–10.5 (7.0–8.0) * 6.0–10.0 (7.0–8.0) 6.0–11.0 (7.0–8.0) 6.0–10.5 (7.0–8.0) 5.5–9.0 (8.5) 6.0–10.5 (7.0–8.0) † 6.0–9.5 (7.5) 5.5–10.5 (8.0–8.5) Oxidase – + + + + – – – + Hydrolysis of: Gelatin – – + W – + – + – Starch – – – + + – + – + Tween 80 + W + – – ND – – – Utilization of: D-glucose – + + + + ND + + – Maltose + + + – – ND + + – Mannitol – + + + + – + – + Enzyme activity (API ZYM): Esterase (C4) – + + * + + + – * + W Lipase esterase (C8) + + + * + – + W * ND W Naphthol-AS-BI-phosphohydrolase – – – * + + – W * ND – Trypsin + – – * + – + – * ND – α -Chymotrypsin + – – * + – + – * ND + β -Galactosidase – – + * – – + + * + – α -Glucosidase – – + * + – + – * ND + Reduction of: Nitrates to nitrites + – + + – ND + – – DNA G + C content (%) 43.0 41.6 * 43.1 42.7 42.6 43.7 41.1 41.6 39.7 Strains: 1, Jeotgalibacillus haloalkaliphilus HH7-29 T (this study); 2, Jeotgalibacillus salarius KCTC 13257 T (this study); 3, Jeotgalibacillus alimentarius YKJ-13 T ; 4, Jeotgalibacillus malaysiensis D5 T ; 5, Jeotgalibacillus terrae DSM 22174 T ; 6, Jeotgalibacillus aurantiacus T12 T ; 7, Jeotgalibacillus campisalis SF-57 T ; 8, Jeotgalibacillus proteolyticus 22-7 T ; 9, Jeotgalibacillus soli P9 T . Data for columns 3–9 taken from Yoon et al. ( 2001 ), Yaakop et al. ( 2015 ), Chen et al. ( 2010 ), Jiang et al. ( 2022 ), Yoon et al. ( 2004 ), Li et al. ( 2018 ), Cunha et al. ( 2012 ), respectively. +, positive; –, negative; W, weakly positive reaction; ND, no data ; *, Data from Yoon et al. 2010 ; †, Data from Li et al. 2018 . Growth and morphological characteristics The cells of strain HH7-29 T were identified as Gram-stain positive, aerobic, rod-shaped (approximately 0.6–0.9 µm wide and 1.0–1.6 µm long) with polar flagella (Fig. S6). The strain was catalase-positive but oxidase-negative. Through antibiotic susceptibility testing, strain HH7-29 T was susceptible to ampicillin, cephalexin, erythromycin, roxithromycin, oxytetracycline, streptomycin, kanamycin, sulfamethoxazole, sulfafurazole, norfloxacin, levofloxacin, vancomycin, polymyxin B, licomycin, chloramphenicol, but resistant to tetracycline. Detailed phenotypic characteristics were listed in the species description. The complete morphological and physiological differences between the species are presented in Table 1 . Chemotaxonomic characteristics The major fatty acids (> 10.0% of the total fatty acids) of strain HH7-29 T were iso-C 15:0 (35.0%) and anteiso-C 15:0 (15.4%). This profile is similar to that of the related species (except for J. campisalis ), but with a rather wide range of abundance (11.0–57.1% and 15.2–53.5%, respectively). Furthermore, strain HH7-29 T differs from the closely related species in terms of the proportions of certain moderate fatty acid components. For example, strain HH7-29 T had significantly lower levels of alcohol-C 17:0 (6.0%) compared with reference strain J. salarius KCTC 13257 T (11.5%) (Table S7). The polar lipids compositions of strain HH7-29 T was identified as phosphatidylglycerol (PG), diphosphatidylglycerol (DPG) and two unidentified phospholipids (PL1 and PL2) (Fig. S7). Therein, PG, DPG and PL2 were detected in J. salarius KCTC 13257 T , but no PL1 (Fig. S7). Furthermore, the main respiratory quinones of strain HH7-29 T were MK-7 and MK-8, in line with all other members of the Jeotgalibacillus representatives. TAXONMIC CONCLUSION The phenotypic, phylogenetic, genomic, physiological and chemotaxonomic features all support the hypothesis that strain HH7-29 T represents a novel species of the genus Jeotgalibacillus , for which the name Jeotgalibacillus haloalkaliphilus sp. nov. is proposed here. DESCRIPTION OF JEOTGALIBACILLUS HALOALKALIPHILUS SP. NOV. Jeotgalibacillus haloalkaliphilus [hal.o.al.ka.li’phi.lus. Gr. masc. n. hals, salt; N.L. neut. n. alkali, alkali (indeclinable); Gr. masc. adj. philos, loving; N.L. masc. adj. haloalkaliphilus, loving briny and alkaline media] Cells are Gram-stain positive, aerobic and rod-shaped (0.6–0.9×1.0–1.6 µm) with polar flagella. Colonies are yellow, circular, raised and approximately 2 mm in diameter on TYS agar after incubation for 2 days. Growth occurs at 15–40℃ (optimum, 32℃), in 0.5–24% NaCl (optimum, 2–9%) and at pH 6.0–12.0 (optimum, pH 8.0–8.5). The strain is catalase-positive and oxidase-negative. Casein, milk, Tweens 20, 40, 60 and 80 are hydrolysed, but starch, gelatin, cellulose and sodium alginate are not hydrolysed. In API 20NE tests, positive for nitrate reduction, urease, arginine dihydrolase, hydrolysis of aesculin, acid production from β -galactosidase and assimilation of maltose, gluconate, malic acid and citric acid, but negative for indole production, hydrolysis of gelatin, acidification of glucose and assimilation of glucose, arabinose, mannose, mannitol, N-acetylglucosamine, capric acid, adipate and phenylacetic acid. In API ZYM tests, there are positive reactions for alkaline phosphatase, esterase lipase (C8), trypsin, and α -chymotrypsin, but negative reactions for esterase (C4), lipase (C14), leucine arylamidase, valine aminopeptidase, cystine aminopeptidase, acid phosphatase, naphthol-AS-BI-phosphohydrolase, α -galactosidase, β -galactocosidase, β -glucuronidase, α -glucosidase, β -glucosinase, α -mannosidase, β -fucosidase, and N-acetyl- β -glucosaminidase. The predominant fatty acids are iso-C 15:0 and anteiso-C 15:0 . The polar lipids consist of phosphatidylglycerol, diphosphatidylglycerol, and two unidentified phospholipids. The major menaquinones are MK-7 and MK-8. The type strain, HH7-29 T (= KCTC 43417 T = MCCC 1K07541 T ), was isolated from the estuary of the Fenhe River into the Yellow River of Shanxi, China. The genomic DNA G + C content of the type strain is 43.0%. The NCBI GenBank accession numbers for the 16S rRNA gene sequence and the draft genome sequence are OR856029 and JAXQNN000000000, respectively. Abbreviations MCCC Marine culture collection of China KCTC Korean collection for type cultures MEGA Molecular evolutionary genetics analysis OCs Orthologous clusters ANI Average nucleotide identity dDDH Digital DNA-DNA hybridization AAI Amino acid identity POCP Percentage of conserved proteins TLC Thin-layer chromatography Declarations The GenBank accession numbers for the 16S rRNA gene sequence and the genome sequence of strain HH7-29 T are OR856029 and JAXQNN000000000, respectively. Author contributions YFZ and YLP designed the experiments and wrote the manuscript. YHX, BY and MYC performed the genome analysis. ZLZ and LQH revised the manuscript. YL supervised the experiments. Funding This work was funded by the National Natural Science Foundation of China (32300115), the Basic Research Program of Shanxi Province (20210302124004), by the Scientific and Technological Innovation Programs of Shanxi Agricultural University (2020BQ39), by the Scientific and Technological Innovation Programs of Higher Education Institutions in Shanxi Province (2021L134) and by the Excellent Talents Come to Shanxi to Reward Scientific Research Projects (SXYBKY2019025).. Data availability All data obtained in this study are shown within the manuscript and supplemental materials. Competing interests The authors have no relevant financial or non-financial interests to disclose. Ethics approval The authors have declared that no ethical issues exist. References Aziz RK, Bartels D, Best AA, DeJongh M, Disz T et al (2008) The RAST Server: rapid annotations using subsystems technology. BMC genomics 9:75 Burg MB, Ferraris JD (2008) Intracellular organic osmolytes: function and regulation. J Biol Chem 283:7309–7313 Chen YG, Peng DJ, Chen QH, Zhang YQ, Tang SK et al (2010) Jeotgalibacillus soli sp. nov., isolated from non-saline forest soil, and emended description of the genus Jeotgalibacillus . Antonie van Leeuwenhoek 98(3):415–421 Chun J, Oren A, Ventosa A, Christensen H, Arahal DR et al (2018) Proposed minimal standards for the use of genome data for the taxonomy of prokaryotes. Int J Syst Evol Microbiol 68(1):461–466 Collins MD, Jones D (1980) Lipids in the classification and identification of coryneform bacteria containing peptidoglycans based on 2, 4-diaminobutyric acid. Appl Bacteriol 48:459–470 Cunha S, Tiago I, Paiva G, Nobre F, da Costa MS et al (2012) Jeotgalibacillus soli sp. nov., a Gram-stain-positive bacterium isolated from soil. Int J Syst Evol Microbiol 62(Pt 3):608–612 Edgar RC (2004) MUSCLE: Multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Res 32:1792-1797 Felsenstein J (1981) Evolutionary trees from DNA sequences: a maximum likelihood approach. J Mol Evol 17:368–376 Ito M, Xu H, Guffanti AA, Wei Y, Zvi L et al (2004) The voltage-gated Na + channel NaVBP has a role in motility, chemotaxis, and pH homeostasis of an alkaliphilic Bacillus. Proc Natl Acad Sci 101(29):10566–10571 Jackman SD, Vandervalk BP, Mohamadi H, Chu J, Yeo S et al (2017) ABySS 2.0: resource-efficient assembly of large genomes using a Bloom filter. Genome Res 27:768–777 Jiang HN, Yun ST, Wang BX, Zhang MJ, Ma Y et al (2022) Jeotgalibacillus aurantiacus sp. nov., a novel orange-pigmented species with a carotenoid biosynthetic gene cluster, isolated from wetland soil. Antonie van Leeuwenhoek 115(6):773–782 Jones DM (1981) Manual of Methods for General Bacteriology. J Clin Pathol 34(9):1069 Jones DT, Taylor WR, Thornton JM (1992) The rapid generation of mutation data matrices from protein sequences. Comput Appl Biosci 8:275–282 Kim M, Oh HS, Park SC, Chun J (2014) Towards a taxonomic coherence between average nucleotide identity and 16S rRNA gene sequence similarity for species demarcation of prokaryotes. Int J Syst Evol Microbiol 64(Pt 2):346–351 Komagata K, Suzuki KI (1987) Lipid and cell wall analysis in bacterial systematics. Methods Microbiol 19:161-207 Koichiro T, Glen S, Sudhir K (2021) MEGA11: molecular evolutionary genetics analysis version 11. Mol Biol Evol 38:3022–3027 Krulwich TA, Hicks DB, Ito M (2009) Cation/proton antiporter complements of bacteria: why so large and diverse? Mol Microbiol 74(2):257–260 Lechner M, Findeiss S, Steiner L, Marz M, Stadler PF et al (2011) Proteinortho: Detection of (Co-)orthologs in large-scale analysis. BMC bioinformatics 12:124 Liew KJ, Lim L, Woo HY, Chan KG, Shamsir MS et al (2018) Purification and characterization of a novel GH1 beta-glucosidase from Jeotgalibacillus malaysiensis . Int J Syst Evol Microbiol 115:1094–1102 Li Y, Zhang Z, Xu Z, Fang D, Wang ET et al (2018) Jeotgalibacillus proteolyticus sp. nov., a protease-producing bacterium isolated from ocean sediments. Int J Syst Evol Microbiol 68(12):3790–3795 Medema MH, Blin K, Cimermancic P, de Jager V, Zakrzewski P et al (2011) antiSMASH: rapid identification, annotation and analysis of secondary metabolite biosynthesis gene clusters in bacterial and fungal genome sequences. Nucleic Acids Res 39:W339–W346 Meier-Kolthoff JP, Auch AF, Klenk HP, Göker M （2013） Genome sequence-based species delimitation with confidence intervals and improved distance functions. BMC Bioinformatics 14:60 Mongodin EF, Nelson KE, Daugherty S, Deboy RT, Wister J et al (2005) The genome of Salinibacter ruber : convergence and gene exchange among hyperhalophilic bacteria and archaea. Proc Natl Acad Sci USA 102:18147–18152 Pardi F, Guillemot S, Gascuel O (2010) Robustness of phylogenetic inference based on minimum evolution. Bull Math Biol 72:1820–1839 Qin QL, Xie BB, Zhang XY, Chen XL, Zhou BC et al (2014) A proposed genus boundary for the prokaryotes based on genomic insights. J Bacteriol 196(12):2210–2215 Rüger HJ, Richter G (1979) Bacillus globisporus subsp. marinus subsp. nov. Int J Syst Evol Microbiol 29:196-203 Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425 Smibert RM, Krieg NR (1981) Manual of methods for general bacteriology. American Society for Microbiology, Washington, DC, pp 409–443 Soliev AB, Hosokawa K, Enomoto K (2011) Bioactive pigments from marine bacteria: applications and physiological roles. Evid Based Complement Alternat Med 2011:1–17 Srinivas A, Divyasree B, Sasikala C, Tushar L, Bharti D et al (2016) Description of Jeotgalibacillus alkaliphilus sp. nov., isolated from a solar salt pan, and Jeotgalibacillus terrae sp. nov., a name to replace ' Jeotgalibacillus soli ' Chen et al . 2010. Int J Syst Evol Microbiol 66(12):5167–5172 Tatusova T, DiCuccio M, Badretdin A, Chetvernin V, Nawrocki EP et al (2016) NCBI prokaryotic genome annotation pipeline. Nucleic Acids Res 44:6614–6624 Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680 Vargas C, Argandoña M, ReinaBueno M, Rodríguez-Moya J, Fernández-Aunión C et al (2008) Unravelling the adaptation responses to osmotic and temperature stress in Chromohalobacter salexigens , a bacterium with broad salinity tolerance. Saline Syst 4:14 Wargo MJ (2013) Homeostasis and catabolism of choline and glycine betaine: lessons from Pseudomonas aeruginosa. Appl Environ Microbiol 79:2112–2120 Weisburg WG, Barns SM, Pelletier DA, Lane DJ (1991) 16S ribosomal DNA amplification for phylogenetic study. J Bacteriol 173:697–703 Yaakop AS, Chan KG, Ee R, Kahar UM, Kon WC et al (2015) Isolation of Jeotgalibacillus malaysiensis sp. nov. from a sandy beach, and emended description of the genus Jeotgalibacillus . Int J Syst Evol Microbiol 65(7):2215–2221 Yaakop AS, Chan KG, Gan HM, Goh KM (2015) Draft genome of Jeotgalibacillus campisalis SF-57(T), a moderate halophilic bacterium isolated from marine saltern. Marine genomics 23:59–60 Yoon JH, Kang SJ, Schumann P, Oh TK (2010) Jeotgalibacillus salarius sp. nov., isolated from a marine saltern, and reclassification of Marinibacillus marinus and Marinibacillus campisalis as Jeotgalibacillus marinus comb. nov. and Jeotgalibacillus campisalis comb. nov., respectively. Int J Syst Evol Microbiol 60(Pt 1):15–20 Yoon JH, Kim IG, Schumann P, Oh TK, Park YH (2004) Marinibacillus campisalis sp. nov., a moderate halophile isolated from a marine solar saltern in Korea, with emended description of the genus Marinibacillus . Int J Syst Evol Microbiol 54(Pt 4):1317–1321 Yoon JH, Weiss N, Lee KC, Lee IS, Kang KH et al (2001) Jeotgalibacillus alimentarius gen. nov., sp. nov., a novel bacterium isolated from jeotgal with L-lysine in the cell wall, and reclassification of Bacillus marinus Rüger 1983. as mMrinibacillus marinus gen nov., comb. nov. Int J Syst Evol Microbiol 51(Pt 6):2087–2093 Yoon SH, Ha SM, Lim JM, Kwon SJ, Chun J (2017) A large-scale evaluation of algorithms to calculate average nucleotide identity. Antonie van Leeuwenhoek. Int J Syst Evol Microbiol 110:1281–1286 Additional Declarations No competing interests reported. Supplementary Files Supplementarymaterials.docx Cite Share Download PDF Status: Published Journal Publication published 01 May, 2024 Read the published version in Antonie van Leeuwenhoek → Version 1 posted Editorial decision: Revision requested 22 Jan, 2024 Editor assigned by journal 22 Jan, 2024 Submission checks completed at journal 20 Jan, 2024 First submitted to journal 19 Jan, 2024 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. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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-3880322","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":268580886,"identity":"8e430f96-370b-4ea0-9601-d31db4f2322f","order_by":0,"name":"Ya-Fei Zhang","email":"","orcid":"","institution":"Shanxi Agricultural University","correspondingAuthor":false,"prefix":"","firstName":"Ya-Fei","middleName":"","lastName":"Zhang","suffix":""},{"id":268580887,"identity":"bd21bc4f-895c-4ab8-949e-bb6eb57e99a9","order_by":1,"name":"Yun-Lin Peng","email":"","orcid":"","institution":"Shanxi Agricultural University","correspondingAuthor":false,"prefix":"","firstName":"Yun-Lin","middleName":"","lastName":"Peng","suffix":""},{"id":268580888,"identity":"a2afa478-e8b3-4e0f-ac76-60b6bb6db1a1","order_by":2,"name":"Yong-Hui Xiao","email":"","orcid":"","institution":"Shanxi Agricultural University","correspondingAuthor":false,"prefix":"","firstName":"Yong-Hui","middleName":"","lastName":"Xiao","suffix":""},{"id":268580889,"identity":"6033654f-8f53-40ea-834e-1dd5cf701092","order_by":3,"name":"Bing Yu","email":"","orcid":"","institution":"Shanxi Agricultural University","correspondingAuthor":false,"prefix":"","firstName":"Bing","middleName":"","lastName":"Yu","suffix":""},{"id":268580890,"identity":"9f248f9a-a5de-4623-8d98-facadbffba4c","order_by":4,"name":"Ming-Yue Cao","email":"","orcid":"","institution":"Shanxi Agricultural University","correspondingAuthor":false,"prefix":"","firstName":"Ming-Yue","middleName":"","lastName":"Cao","suffix":""},{"id":268580891,"identity":"f3534fdb-c507-46c6-a178-4815442b66a0","order_by":5,"name":"Zhi-Ling Zhang","email":"","orcid":"","institution":"Shanxi Agricultural University","correspondingAuthor":false,"prefix":"","firstName":"Zhi-Ling","middleName":"","lastName":"Zhang","suffix":""},{"id":268580892,"identity":"0c55f2bc-35cd-4abe-87f4-9a16726d4d82","order_by":6,"name":"Le-Qun Huang","email":"","orcid":"","institution":"Shanxi Agricultural University","correspondingAuthor":false,"prefix":"","firstName":"Le-Qun","middleName":"","lastName":"Huang","suffix":""},{"id":268580893,"identity":"1550671d-df25-4b19-be3f-0c397e984d3c","order_by":7,"name":"Yi Li","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAtElEQVRIiWNgGAWjYBACPmYGhgMfoBwJorSwAbUcnEGaFiBm5iFNCzuP4WGbP4flDQ4wH7zNw2CXR4TDeAwO57alGW44wJZszcOQXEyklgYbxg0HeMykeRgOJDYQpcXij4T9hgP830jQwsBmkwi0hY1YLWwFB3vb0pJnHmYztpxjkExYCz//4c0ffvw5bNt3vPnhjTcVdoS1MDBwGEBoZhBhQFg9ELA/IErZKBgFo2AUjGAAAKmdNMD0M6tTAAAAAElFTkSuQmCC","orcid":"","institution":"Shanxi Agricultural University","correspondingAuthor":true,"prefix":"","firstName":"Yi","middleName":"","lastName":"Li","suffix":""}],"badges":[],"createdAt":"2024-01-20 02:00:06","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-3880322/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-3880322/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s10482-024-01968-8","type":"published","date":"2024-05-01T19:58:30+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":50034678,"identity":"ccc3baf8-2d98-4ff8-9065-9cfeea60a8d5","added_by":"auto","created_at":"2024-01-23 13:32:49","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":388323,"visible":true,"origin":"","legend":"\u003cp\u003eMaximum-likelihood phylogenetic tree based on the 16S rRNA gene sequences, showing the phylogenetic positions of strain HH7-29\u003csup\u003eT\u003c/sup\u003e and related species in the order \u003cem\u003eJeotgalibacillus\u003c/em\u003e. Bootstrap values (\u0026gt;50%) based on 1000 replicates are shown at nodes. Filled circles indicate branches that were also recovered in both the neighbor­joining tree and the minimum-evolution tree. Bar, 0.005 substitutions per nucleotide position.\u003c/p\u003e","description":"","filename":"Figure1.png","url":"https://assets-eu.researchsquare.com/files/rs-3880322/v1/73b1fc8257b474ebceba8d94.png"},{"id":50034679,"identity":"d4f9add6-c486-4c9b-89c8-f73fe9a231f3","added_by":"auto","created_at":"2024-01-23 13:32:49","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":539526,"visible":true,"origin":"","legend":"\u003cp\u003eComparison of gene contents of strain HH7-29\u003csup\u003eT\u003c/sup\u003e and its related species. The Venn diagram shows the core and specific genes in each strain.\u003c/p\u003e","description":"","filename":"Figure2.png","url":"https://assets-eu.researchsquare.com/files/rs-3880322/v1/43caa2a7bfb4cbad801297fa.png"},{"id":50034681,"identity":"f73f0816-3606-4ae8-90f5-fd8a1afdae7f","added_by":"auto","created_at":"2024-01-23 13:32:49","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":356490,"visible":true,"origin":"","legend":"\u003cp\u003eMaximum-likelihood phylogenetic tree based on 815 single-copy orthologous cluster sequences from genome sequences showing phylogenetic positions of strain HH7-29\u003csup\u003eT\u003c/sup\u003e and related species in the family \u003cem\u003eJeotgalibacillus\u003c/em\u003e. Filled circles indicate branches that were also recovered in both the neighbor­joining tree and the minimum-evolution tree. Bar, 0.05 substitutions per amino acid position.\u003c/p\u003e","description":"","filename":"Figure3.png","url":"https://assets-eu.researchsquare.com/files/rs-3880322/v1/5227a2431f401c1e5731573f.png"},{"id":50034958,"identity":"08930226-76bb-4288-803f-e73a69cdba75","added_by":"auto","created_at":"2024-01-23 13:40:49","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":417643,"visible":true,"origin":"","legend":"\u003cp\u003eNormalized distribution of pI values at 0.4 intervals for predicted proteins in HH7-29\u003csup\u003eT\u003c/sup\u003e, the closely related strains, hyperhalophilic bacterium \u003cem\u003eSalinibacter ruber\u003c/em\u003e DSM 13855\u003csup\u003eT\u003c/sup\u003e, non-halophilic bacterium \u003cem\u003eEscherichia coli\u003c/em\u003e K-12\u003csup\u003eT\u003c/sup\u003e. Predicted pI values of the proteins were calculated by the EMBOSS Pepstats program.\u003c/p\u003e","description":"","filename":"Figure4.png","url":"https://assets-eu.researchsquare.com/files/rs-3880322/v1/3a9379e35f9e1734caf4f5e4.png"},{"id":56042943,"identity":"cabb777c-f4f5-4fba-bcff-bc591395fd13","added_by":"auto","created_at":"2024-05-07 20:09:18","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2455683,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3880322/v1/db5c5fc2-901b-4e06-a81b-8455efda57f1.pdf"},{"id":50034683,"identity":"a0741678-b516-44ec-b298-b68e70657ba3","added_by":"auto","created_at":"2024-01-23 13:32:51","extension":"docx","order_by":6,"title":"","display":"","copyAsset":false,"role":"supplement","size":18078252,"visible":true,"origin":"","legend":"","description":"","filename":"Supplementarymaterials.docx","url":"https://assets-eu.researchsquare.com/files/rs-3880322/v1/bb42beb046a3c9a0cabacd60.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Jeotgalibacillus haloalkaliphilus sp. nov., a novel alkaliphilic and halotolerant bacterium, isolated from the estuary of the Fenhe River into the Yellow River","fulltext":[{"header":"INTRODUCTION","content":"\u003cp\u003eThe genus \u003cem\u003eJeotgalibacillus\u003c/em\u003e, a member of the family \u003cem\u003eCaryophanaceae\u003c/em\u003e within the order \u003cem\u003eCaryophanales\u003c/em\u003e of the class \u003cem\u003eBacilli\u003c/em\u003e, was first identified by Yoon \u003cem\u003eet al\u003c/em\u003e in 2001 with the description of \u003cem\u003eJeotgalibacillus alimentarius\u003c/em\u003e as the type species (Yoon et al. \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2001\u003c/span\u003e). At the time of writing, the genus \u003cem\u003eJeotgalibacillus\u003c/em\u003e comprised 10 recognized species: \u003cem\u003eJeotgalibacillus alimentarius\u003c/em\u003e (Yoon et al. \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2001\u003c/span\u003e), \u003cem\u003eJeotgalibacillus alkaliphilus\u003c/em\u003e (Srinivas et al. \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2016\u003c/span\u003e), \u003cem\u003eJeotgalibacillus aurantiacus\u003c/em\u003e (Jiang et al. \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2022\u003c/span\u003e), \u003cem\u003eJeotgalibacillus campisalis\u003c/em\u003e (Yoon et al. \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2004\u003c/span\u003e), \u003cem\u003eJeotgalibacillus malaysiensis\u003c/em\u003e (Yaakop et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2015\u003c/span\u003e), \u003cem\u003eJeotgalibacillus marinus\u003c/em\u003e (R\u0026uuml;ger and Richter \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e1979\u003c/span\u003e), \u003cem\u003eJeotgalibacillus proteolyticus\u003c/em\u003e (Li et al. \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2018\u003c/span\u003e), \u003cem\u003eJeotgalibacillus salarius\u003c/em\u003e (Yoon et al. \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2010\u003c/span\u003e), \u003cem\u003eJeotgalibacillus soli\u003c/em\u003e (Cunha et al. \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2012\u003c/span\u003e) and \u003cem\u003eJeotgalibacillus terrae\u003c/em\u003e (Srinivas et al. \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2016\u003c/span\u003e), among which \u003cem\u003eJeotgalibacillus campisalis\u003c/em\u003e and \u003cem\u003eJeotgalibacillus marinus\u003c/em\u003e were named through the reclassification of two \u003cem\u003eMarinibacillus\u003c/em\u003e species (Yoon et al. \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). Except for \u003cem\u003eJ. terrae\u003c/em\u003e and \u003cem\u003eJ. aurantiacus\u003c/em\u003e (from forest soil and wetland soil, respectively), the other species originated from saline or alkaline environments, such as marine saltern, salt pan, salted fermented food and alkaline sandy soil. These \u003cem\u003eJeotgalibacillus\u003c/em\u003e species represent a group of Gram-stain-positive, rod-shaped bacteria with DNA G\u0026thinsp;+\u0026thinsp;C content of 39.3\u0026ndash;44.0%, containing MK-7 and MK-8 as the major quinones, and iso-C\u003csub\u003e15:0\u003c/sub\u003e and anteiso-C\u003csub\u003e15:0\u003c/sub\u003e as the major cellular fatty acids (Yaakop et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). Most of members exhibit a yellow pigmentation (Jiang et al. \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2022\u003c/span\u003e), which may be associated with their self-defense against harsh environmental conditions (Soliev et al. \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2011\u003c/span\u003e). Some species, such as \u003cem\u003eJ. malaysiensis\u003c/em\u003e and \u003cem\u003eJ. proteolyticus\u003c/em\u003e, have ability for secreting a multitude of enzymes (e.g., protease, gelatinase, esterase and \u003cem\u003eβ\u003c/em\u003e-glucosidase), which have important implications towards the biotechnological industry (Liew et al. \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Li et al. \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2018\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eHigh salinity is one of the most challenging stresses for life on Earth. However, microorganisms have developed two mechanisms to maintain their cellular osmotic balance: by increasing the ion concentrations or by accumulating protective organic osmolytes in cells (Mongodin et al. \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2005\u003c/span\u003e; Burg and Ferraris \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2008\u003c/span\u003e). \u003cem\u003eJeotgalibacillus\u003c/em\u003e spp. were reported to be slightly halophilic, and some of these strains tolerate salt concentrations of up to 30% (w/v) (Yaakop et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). Earlier genomic analyses revealed that the genus \u003cem\u003eJeotgalibacillus\u003c/em\u003e features diverse genes resistant to salt, which help them improve their ecological fitness in alkaline saline ecosystems (Jiang et al. \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). During our survey of the diversity of the culturable bacteria from the river confluences of Shanxi, China, we discovered a novel alkaliphilic and halotolerant bacterium belonging to the genus \u003cem\u003eJeotgalibacillus\u003c/em\u003e, designated as HH7-29\u003csup\u003eT\u003c/sup\u003e. To better understand and use it, strain HH7-29\u003csup\u003eT\u003c/sup\u003e was subjected to polyphasic taxonomic analyses involving phylogenetic, phenotypic and chemotaxonomic profiles. Furthermore, we also analyzed the genomic characteristics associated with alkaline saline tolerance to understand how it addressed the challenges of osmolality fluctuations.\u003c/p\u003e"},{"header":"METHODS","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eStrain isolation and culture conditions\u003c/h2\u003e \u003cp\u003eFresh sediment samples were obtained from the estuary of the Fenhe River into the Yellow River (35\u0026deg;35\u0026prime;N, 110\u0026deg;47\u0026prime;E) of Shanxi, China. 1 g of thoroughly homogenized samples were tenfold serially diluted using sterile 1% NaCl, and spread onto 1/3 MA agar (MA; Difco). Single orange-yellow colony approximately 2 mm in diameter was picked and designated HH7-29\u003csup\u003eT\u003c/sup\u003e. Single colony was transferred into TYS broth (0.5% tryptone, 0.1% yeast extract, 3.0% salt) for growth at 30℃ and stored at \u0026minus;\u0026thinsp;80℃ as glycerol suspensions (20%). For comparative study, the reference strain \u003cem\u003eJeotgalibacillus salarius\u003c/em\u003e KCTC 13257\u003csup\u003eT\u003c/sup\u003e was obtained from the KCTC (Korean Collection for Type Cultures) and grown on TYS agar at 30\u0026deg;C.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e16S rRNA gene sequencing and phylogenetic analysis\u003c/h2\u003e \u003cp\u003eFor phylogenetic analysis of strain HH7-29\u003csup\u003eT\u003c/sup\u003e, the almost-complete 16S rRNA gene sequence was amplified by PCR with primers 27F and 1492R (Weisburg et al. \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e1991\u003c/span\u003e). Following purification, the PCR products were ligated to the pMD19-T Vector (TaKaRa) and sequenced at Sangon Biotech (Shanghai, China) using an ABI 3730 DNA analyser. The sequence of strain HH7-29\u003csup\u003eT\u003c/sup\u003e was compared to reference strains using EzBioCloud database. A multiple alignment of all the sequences was achieved by using CLUSTAL W algorithm (Thompson et al. \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e1994\u003c/span\u003e) integrated in MEGA 11 (Koichiro et al. \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Phylogenetic tree was reconstructed with MEAG 11 using the maximum-likelihood (ML) (Felsenstein \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e1981\u003c/span\u003e), neighbour-joining (NJ) (Saitou and Nei \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e1987\u003c/span\u003e), and minimum-evolution (ME) (Pardi et al. \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2010\u003c/span\u003e) algorithm, and evaluated by bootstrap analysis with 1000 replications.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003eDraft genome sequencing and genome sequences analysis\u003c/h2\u003e \u003cp\u003eWhole genome sequencing of strain HH7-29\u003csup\u003eT\u003c/sup\u003e was conducted by Oebiotech (Shanghai, China) using the Illumina HiSeq 4000 system (Illumina, USA). The paired-end reads assembly were performed with ABySS v2.0 (Jackman et al. \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). Genome annotation was performed with National Center for Biotechnology Information (NCBI) Prokaryotic Genome Annotation Pipeline (PGAP) (Tatusova et al. 2016) and the Rapid Annotations using Subsystems Technology (RAST) server (Aziz et al. \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2008\u003c/span\u003e). The secondary metabolite biosynthesis gene clusters of genome sequences were annotated using antiSMASH bacterial version (Medema et al. \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2011\u003c/span\u003e). The G\u0026thinsp;+\u0026thinsp;C content of the genomic DNA was determined based on the genome sequence. Genomes of closely related strains were retrieved from the NCBI GenBank database. The 815 single-copy orthologous clusters (OCs) (Jones et al. \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e1992\u003c/span\u003e) were selected using Proteinortho v5.16 (Lechner et al. \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2011\u003c/span\u003e) and aligned separately with MUSCLE v3.8.31 (Edgar \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2004\u003c/span\u003e). Phylogenetic trees based on the whole genome were reconstructed using MEGA 11. Average nucleotide identity (ANI) values were calculated using EzBioCloud web service (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.ezbiocloud.net/tools/ani\u003c/span\u003e\u003cspan address=\"https://www.ezbiocloud.net/tools/ani\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e) (Yoon et al. \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2017\u003c/span\u003e), and digital DNA\u0026ndash;DNA hybridization (dDDH) values were determined using the Genome-to-Genome Distance Calculator (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://ggdc.dsmz.de/ggdc.php\u003c/span\u003e\u003cspan address=\"http://ggdc.dsmz.de/ggdc.php\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e) (Meier-Kolthoff et al. \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). Average amino acid identity (AAI) values were calculated using an online AAI calculator server (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://enve-omics.ce.gatech.edu/aai/\u003c/span\u003e\u003cspan address=\"http://enve-omics.ce.gatech.edu/aai/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e) (Luo et al. 2014), and the percentage of conserved proteins (POCP) values were estimated based on a previously described approach (Qin et al. \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2014\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003eGrowth and morphological characteristics\u003c/h2\u003e \u003cp\u003eThe morphological of strain HH7-29\u003csup\u003eT\u003c/sup\u003e was investigated after cultivation on TYS at 30℃ for 12 h. Cell size, shape and flagella were assessed with transmission electron microscopy (Tecnai G2 F20, FEI). The Gram reaction was tested using a Gram-stain kit (Solarbio, China) according to the manufacturer\u0026rsquo;s instructions. Temperature sensitivity was determined in TYS broth over a range of 4\u0026ndash;45℃ (4, 10, 15, 20, 25, 28, 32, 37, 40 and 45℃). Salt tolerance was tested by using a modified medium (0.5% tryptone, 0.1% yeast extract, 0.5% MgCl\u003csub\u003e2\u003c/sub\u003e, 0.05% CaCl\u003csub\u003e2\u003c/sub\u003e, 0.1% KCl and 0.0001% FeSO\u003csub\u003e4\u003c/sub\u003e) with varying NaCl concentrations (0\u0026ndash;4% NaCl, increased at intervals of 0.5%; 4\u0026ndash;30% NaCl, increased at intervals of 1%). The pH range for growth was tested in TYS broth at pH 5.0\u0026thinsp;\u0026minus;\u0026thinsp;10.0 (increments of 0.5 pH unit) by adding 50 mmol/L buffers [MES (pH 5.0\u0026thinsp;\u0026minus;\u0026thinsp;6.0), MOPS (pH 6.5\u0026thinsp;\u0026minus;\u0026thinsp;7.0), Tris (pH 7.5\u0026thinsp;\u0026minus;\u0026thinsp;8.5) and CHES (pH 9.0\u0026thinsp;\u0026minus;\u0026thinsp;10.0)]. Catalase activity was tested by assessing the formation of bubbles upon the addition of 3% (w/v) H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e (Jones \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e1981\u003c/span\u003e). Oxidase activity was performed with oxidase strips (Merck, Germany). Anaerobic respiration was determined in Hungate tubes filled with TYS broth supplemented with 0.1% (w/v) potassium nitrate, 0.05% (w/v) cysteine hydrochloride and 0.05% (w/v) sodium sulfide. Hydrolyses of casein (0.5%, w/v), starch (0.5%, w/v), milk (0.5%, w/v), gelatin (0.5%, w/v), cellulose (0.5%, w/v), sodium alginate (0.5%, w/v) and Tweens (20, 40, 60 and 80, 1%, v/v) were tested as previously described by Smibert and Krieg (Smibert and Krieg \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e1981\u003c/span\u003e). Antibiotic susceptibility testing was performed according to the antibiotic disc-difusion method using 16 common antibiotics: ampicillin (10 \u0026micro;g), cephalexin (30 \u0026micro;g), erythromycin (15 \u0026micro;g), roxithromycin (15 \u0026micro;g), tetracycline (30 \u0026micro;g), oxytetracycline (30 \u0026micro;g), streptomycin (10 \u0026micro;g), kanamycin (30 \u0026micro;g), sulfamethoxazole (25 \u0026micro;g), sulfafurazole (25 \u0026micro;g), norfloxacin (10 \u0026micro;g), levofloxacin (5 \u0026micro;g), vancomycin (30 \u0026micro;g), polymyxin B (300 IU), licomycin (2 \u0026micro;g), and chloramphenicol (30 \u0026micro;g). Other physiological and biochemical characteristics were determined with API 20 NE and API ZYM systems (bioM\u0026eacute;rieux, France) following the manufacture instructions.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003eChemotaxonomic analysis\u003c/h2\u003e \u003cp\u003eFor the analyses of fatty acids, respiratory quinones, and polar lipids, strain HH7-29\u003csup\u003eT\u003c/sup\u003e and the reference strain \u003cem\u003eJ. salarius\u003c/em\u003e KCTC 13257\u003csup\u003eT\u003c/sup\u003e were obtained from cultures grown in TYS broth at 30℃ for 12 h (logarithmic phase of growth). The harvested cells were washed twice in sterile 1% NaCl and then freeze dried. Cellular fatty acids were extracted, methylated, and identified using gas chromatography (Agilent 6850), following the instructions of the Sherlock Microbial Identification System (MIDI, version 6.1). Peaks were automatically integrated, and fatty acid names and percentages were determined using the Microbial Identification System (MIS) standard software. The polar lipids were extracted according to Komagata and Suzuki (\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e1987\u003c/span\u003e) and determined using a 2D thin-layer chromatography (TLC) procedure on Merck silica gel 60 F254 plates. The staining reagents phosphomolybdic acids was used for detecting the total lipids, ninhydrin for lipids containing free amino groups, and molybdenum blue for phospholipids (Collins and Jones \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e1980\u003c/span\u003e). Respiratory quinones were extracted from freeze-dried cells using a mixture of chloroform/methanol (2:1, v/v), and the extract was analyzed by an LC\u0026ndash;MS system (a Dionex Ultimate 3000 HPLC device connected to a Bruker impact HD mass spectrometer).\u003c/p\u003e \u003c/div\u003e"},{"header":"RESULTS AND DISCUSSION","content":"\u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003ePhylogenetic analysis based on 16S rRNA gene sequences\u003c/h2\u003e \u003cp\u003eThe nearly full-length 16S rRNA gene sequence of strain HH7-29\u003csup\u003eT\u003c/sup\u003e (1513 bp) has been deposited as OR856029 in the GenBank/ database. On pairwise comparison with the 16S rRNA gene sequences available in the EzBioCloud database, strain HH7-29\u003csup\u003eT\u003c/sup\u003e exhibited the highest sequence similarities to \u003cem\u003eJ. alkaliphilus\u003c/em\u003e JC303\u003csup\u003eT\u003c/sup\u003e (98.4%), followed by \u003cem\u003eJ. salarius\u003c/em\u003e ASL-1\u003csup\u003eT\u003c/sup\u003e (98.1%), \u003cem\u003eJ. alimentarius\u003c/em\u003e YKJ-13\u003csup\u003eT\u003c/sup\u003e (98.1%) and other \u003cem\u003eJeotgalibacillus\u003c/em\u003e members (94.6\u0026ndash;98.0%). These values were lower than the threshold (98.7%) for separating different species (Chun et al. \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). In the ML, NJ and ME phylogenetic trees (Figs.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e, S1 and S2), strain HH7-29\u003csup\u003eT\u003c/sup\u003e fell within a sub-cluster of the genus \u003cem\u003eJeotgalibacillus\u003c/em\u003e, which consisted of five \u003cem\u003eJeotgalibacillus\u003c/em\u003e species (\u003cem\u003eJ. alkaliphilus\u003c/em\u003e, \u003cem\u003eJ. salarius\u003c/em\u003e, \u003cem\u003eJ. malaysiensis\u003c/em\u003e, \u003cem\u003eJ. alimentarius\u003c/em\u003e and \u003cem\u003eJ. terrae\u003c/em\u003e). Within this sub-cluster, strain HH7-29\u003csup\u003eT\u003c/sup\u003e formed an internally independent branch among adjacent strains. The taxonomic identification and evolutionary tree based on the 16S rRNA gene showed that strain HH7-29\u003csup\u003eT\u003c/sup\u003e represented a potential novel species.\u003c/p\u003e\u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003eGenome features and comparison\u003c/h2\u003e \u003cp\u003eThe assembled genome sequencing of strain HH7-29\u003csup\u003eT\u003c/sup\u003e (JAXQNN000000000) yielded a genome of 3284775 bp in length, which was composed of 28 contigs with an N50 value of 985906 bp (Table \u003cspan refid=\"MOESM1\" class=\"InternalRef\"\u003eS1\u003c/span\u003e). The G\u0026thinsp;+\u0026thinsp;C content was 43.0%, which was in the range of 39.7\u0026ndash;43.7% for other species of \u003cem\u003eJeotgalibacillus\u003c/em\u003e. A total of 3337 genes were predicted in the genome, including 3265 protein-coding genes, 16 pseudo genes and 56 RNA genes (6 rRNAs, 5 ncRNAs and 45 tRNAs). 1323 (39.6%) of all predicted genes were assigned through the RAST database. The abundance of carbohydrates metabolism (15.3%), protein metabolism (13.0%) and DNA metabolism (4.7%) in strain HH7-29\u003csup\u003eT\u003c/sup\u003e fell within the range of the closely related species (Table S2). While numbers of genes associated with respiration and potassium metabolism in strain HH7-29\u003csup\u003eT\u003c/sup\u003e were higher than that in closely related species. Meanwhile, the genes encoding hydrogenase were unique to strain HH7-29\u003csup\u003eT\u003c/sup\u003e. Additionally, genomic comparisons showed that all the nine strains shared 1352 core genes in common, accounting for 37.3\u0026ndash;42.1% of the annotated genome repertoire respectively (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). The antiSMASH results showed that strain HH7-29\u003csup\u003eT\u003c/sup\u003e harbors two sets of gene clusters required for carotenoid biosynthesis and siderophore biosynthesis, respectively. Additionally, the genes encoding peptidases such as ATP-dependent protease La, aminopeptidase and carboxylate peptidase were detected in the genome. In line with this, strain HH7-29\u003csup\u003eT\u003c/sup\u003e had milk and casein degradation capacity (Fig. S3).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eIn the phylogenetic trees based on 815 single-copy orthologous clusters (Figs.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e, S4 and S5), strain HH7-29\u003csup\u003eT\u003c/sup\u003e clustered within the clade that accommodated the genus \u003cem\u003eJeotgalibacillus\u003c/em\u003e, in which it formed a cluster with \u003cem\u003eJ. malaysiensis\u003c/em\u003e D5\u003csup\u003eT\u003c/sup\u003e and \u003cem\u003eJ. salarius\u003c/em\u003e ASL-1\u003csup\u003eT\u003c/sup\u003e. The ANI values between strain HH7-29\u003csup\u003eT\u003c/sup\u003e and related \u003cem\u003eJeotgalibacillus\u003c/em\u003e species were 71.1\u0026ndash;83.8% (Table S3). Moreover, the dDDH values between strain HH7-29\u003csup\u003eT\u003c/sup\u003e and related species ranged from 19.5 to 27.4% (Table S3). These values are significantly lower than the threshold values for prokaryotic species delineation, which are 95\u0026ndash;96% for ANI and 70% for dDDH (Kim et al. \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Meier-Kolthoff et al. \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). Furthermore, the AAI and POCP values between strain HH7-29\u003csup\u003eT\u003c/sup\u003e and related species were 66.5\u0026ndash;88.4% and 59.8\u0026ndash;76.6% (Table S4), which were within the range indicating the delimitation boundaries of the genus (above 65\u0026ndash;95% and 50%, respectively) (Luo et al. 2014; Qin et al. \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). These results confirmed that HH7-29\u003csup\u003eT\u003c/sup\u003e represents a novel species within the genus \u003cem\u003eJeotgalibacillus\u003c/em\u003e.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eSalinity tolerance\u003c/h2\u003e \u003cp\u003eUnder high-salinity conditions, the growth of microorganisms is severely limited. Surprisingly, strain HH7-29\u003csup\u003eT\u003c/sup\u003e could grow in NaCl concentrations up to 24%, showing high salinity tolerance ability (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). It has been reported that proteins of halophilic organisms have more acidic residues and lower isoelectric points (pIs) to accommodate high ionic strength (Mongodin et al. \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2005\u003c/span\u003e). In this study, through comparison of pIs from HH7-29\u003csup\u003eT\u003c/sup\u003e, the closely related strains, hyperhalophilic bacterium \u003cem\u003eSalinibacter ruber\u003c/em\u003e DSM 13855\u003csup\u003eT\u003c/sup\u003e, non-halophilic bacterium \u003cem\u003eEscherichia coli\u003c/em\u003e K-12\u003csup\u003eT\u003c/sup\u003e, these \u003cem\u003eJeotgalibacillus\u003c/em\u003e members exhibited strongly unimodal distributions of pI values and higher ratios of acidic residues to basic residues, \u003cem\u003eS. ruber\u003c/em\u003e too (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). The average pI values of all proteins in HH7-29\u003csup\u003eT\u003c/sup\u003e and the closely related strains was 6.5, significantly lower than protein pIs of \u003cem\u003eE. coli\u003c/em\u003e K-12\u003csup\u003eT\u003c/sup\u003e with 7.1 (Table S5). Through functional annotation of genes, strain HH7-29\u003csup\u003eT\u003c/sup\u003e possesses betaine/carnitine/choline transporter, betaine/proline/choline transporter and glycine betaine transporter (Table S6). As osmoprotection, compatible solutes (e.g., glycine betaine, proline) accumulation could increase to maintain cellular osmotic balance in saline conditions (Mongodin et al. \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2005\u003c/span\u003e; Burg and Ferraris \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2008\u003c/span\u003e). Though the genes encoding choline dehydrogenases and betaine aldehyde dehydrogenase, which are responsible for synthesizing glycine betaine from choline (Vargas et al. \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2008\u003c/span\u003e; Wargo \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2013\u003c/span\u003e), were lacked, HH7-29\u003csup\u003eT\u003c/sup\u003e could encode two glutamate synthases (UFB30_01585, UFB30_01590). As an organic osmolyte, glutamate could mediate a global transcription switch at genomic promoters that vary in their response to salt (Burg and Ferraris \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2008\u003c/span\u003e). Besides, the genome contained 13 sodium/proton antiporters, five Trk system potassium uptake protein, three sodium/calcium antiporter and two potassium/proton antiporters (Table S6), which maintained high intracellular ion concentrations within cells to achieve osmotic equilibrium. Furthermore, those sodium or potassium/proton antiporters may catalyze proton accumulation coupled with sodium efflux, thereby maintaining a cytoplasmic pH well below the high external pH in strain HH7-29\u003csup\u003eT\u003c/sup\u003e, supporting cytoplasmic pH homeostasis and alkali tolerance(Ito et al. \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2004\u003c/span\u003e; Krulwich et al. \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2009\u003c/span\u003e). Collectively, strain HH7-29\u003csup\u003eT\u003c/sup\u003e could not only accumulate protective organic osmolytes but also increase intracellular ion concentrations to maintain its tolerance to salt, alkali and osmolarity fluctuations.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eDifferential characteristics of strain HH7-29\u003csup\u003eT\u003c/sup\u003e and type strains of closely related species.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"10\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCharacteristic\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c10\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGrowth range of:\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTemperature (optimum, ℃)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e15\u0026ndash;40℃\u003c/p\u003e \u003cp\u003e(32℃)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4\u0026ndash;40℃\u003c/p\u003e \u003cp\u003e(30℃)\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e10\u0026ndash;45℃\u003c/p\u003e \u003cp\u003e(30\u0026ndash;35℃)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e4\u0026ndash;50℃\u003c/p\u003e \u003cp\u003e(37℃)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e10\u0026ndash;45℃\u003c/p\u003e \u003cp\u003e(30\u0026ndash;35℃)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e15\u0026ndash;40℃\u003c/p\u003e \u003cp\u003e(37℃)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e4\u0026ndash;39℃\u003c/p\u003e \u003cp\u003e(30℃)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e10\u0026ndash;40℃\u003c/p\u003e \u003cp\u003e(33℃)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e15\u0026ndash;40℃\u003c/p\u003e \u003cp\u003e(30\u0026ndash;37℃)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNaCl (optimum, %)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.5\u0026ndash;24%\u003c/p\u003e \u003cp\u003e(2\u0026ndash;9%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0\u0026ndash;18%\u003c/p\u003e \u003cp\u003e(2%)\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0\u0026ndash;20%\u003c/p\u003e \u003cp\u003e(3\u0026ndash;12%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0\u0026ndash;30%\u003c/p\u003e \u003cp\u003e(10%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e0\u0026ndash;20%\u003c/p\u003e \u003cp\u003e(2\u0026ndash;5%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0\u0026ndash;9%\u003c/p\u003e \u003cp\u003e(2%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0\u0026ndash;15%\u003c/p\u003e \u003cp\u003e(2\u0026ndash;3%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0\u0026ndash;10%\u003c/p\u003e \u003cp\u003e(2%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e0\u0026ndash;9%\u003c/p\u003e \u003cp\u003e(0\u0026ndash;1%)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003epH (optimum)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e6.0\u0026ndash;12.0\u003c/p\u003e \u003cp\u003e(8.0\u0026ndash;8.5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e6.0\u0026ndash;10.5\u003c/p\u003e \u003cp\u003e(7.0\u0026ndash;8.0)\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e6.0\u0026ndash;10.0\u003c/p\u003e \u003cp\u003e(7.0\u0026ndash;8.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e6.0\u0026ndash;11.0 (7.0\u0026ndash;8.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e6.0\u0026ndash;10.5 (7.0\u0026ndash;8.0)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e5.5\u0026ndash;9.0 (8.5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e6.0\u0026ndash;10.5\u003c/p\u003e \u003cp\u003e(7.0\u0026ndash;8.0)\u003csup\u003e\u0026dagger;\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e6.0\u0026ndash;9.5 (7.5)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e5.5\u0026ndash;10.5 (8.0\u0026ndash;8.5)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOxidase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHydrolysis of:\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGelatin\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eW\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eStarch\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTween 80\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eW\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eND\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eUtilization of:\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eD-glucose\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eND\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMaltose\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eND\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMannitol\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eEnzyme activity (API ZYM):\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eEsterase (C4)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u0026ndash;\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eW\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLipase esterase (C8)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eW\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eND\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003eW\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNaphthol-AS-BI-phosphohydrolase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u0026ndash;\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eW\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eND\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTrypsin\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u0026ndash;\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u0026ndash;\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eND\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eα\u003c/em\u003e-Chymotrypsin\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u0026ndash;\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u0026ndash;\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eND\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eβ\u003c/em\u003e-Galactosidase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e+\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eα\u003c/em\u003e-Glucosidase\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u0026ndash;\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003eND\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eReduction of:\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNitrates to nitrites\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eND\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDNA G\u0026thinsp;+\u0026thinsp;C content (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e43.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e41.6\u003csup\u003e*\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e43.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e42.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e42.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e43.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e41.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e41.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e39.7\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"10\"\u003eStrains: 1, \u003cem\u003eJeotgalibacillus haloalkaliphilus\u003c/em\u003e HH7-29\u003csup\u003eT\u003c/sup\u003e (this study); 2, \u003cem\u003eJeotgalibacillus salarius\u003c/em\u003e KCTC 13257\u003csup\u003eT\u003c/sup\u003e (this study); 3, \u003cem\u003eJeotgalibacillus alimentarius\u003c/em\u003e YKJ-13\u003csup\u003eT\u003c/sup\u003e; 4, \u003cem\u003eJeotgalibacillus malaysiensis\u003c/em\u003e D5\u003csup\u003eT\u003c/sup\u003e; 5, \u003cem\u003eJeotgalibacillus terrae\u003c/em\u003e DSM 22174\u003csup\u003eT\u003c/sup\u003e; 6, \u003cem\u003eJeotgalibacillus aurantiacus\u003c/em\u003e T12\u003csup\u003eT\u003c/sup\u003e; 7, \u003cem\u003eJeotgalibacillus campisalis\u003c/em\u003e SF-57\u003csup\u003eT\u003c/sup\u003e; 8, \u003cem\u003eJeotgalibacillus proteolyticus\u003c/em\u003e 22-7\u003csup\u003eT\u003c/sup\u003e; 9, \u003cem\u003eJeotgalibacillus soli\u003c/em\u003e P9\u003csup\u003eT\u003c/sup\u003e.\u003c/td\u003e\u003c/tr\u003e \u003ctr\u003e\u003ctd colspan=\"10\"\u003eData for columns 3\u0026ndash;9 taken from Yoon et al. (\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2001\u003c/span\u003e), Yaakop et al. (\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2015\u003c/span\u003e), Chen et al. (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2010\u003c/span\u003e), Jiang et al. (\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2022\u003c/span\u003e), Yoon et al. (\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2004\u003c/span\u003e), Li et al. (\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2018\u003c/span\u003e), Cunha et al. (\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2012\u003c/span\u003e), respectively. +, positive; \u0026ndash;, negative; W, weakly positive reaction; ND, no data ; *, Data from Yoon et al. \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; \u0026dagger;, Data from Li et al. \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2018\u003c/span\u003e.\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003eGrowth and morphological characteristics\u003c/h2\u003e \u003cp\u003eThe cells of strain HH7-29\u003csup\u003eT\u003c/sup\u003e were identified as Gram-stain positive, aerobic, rod-shaped (approximately 0.6\u0026ndash;0.9 \u0026micro;m wide and 1.0\u0026ndash;1.6 \u0026micro;m long) with polar flagella (Fig. S6). The strain was catalase-positive but oxidase-negative. Through antibiotic susceptibility testing, strain HH7-29\u003csup\u003eT\u003c/sup\u003e was susceptible to ampicillin, cephalexin, erythromycin, roxithromycin, oxytetracycline, streptomycin, kanamycin, sulfamethoxazole, sulfafurazole, norfloxacin, levofloxacin, vancomycin, polymyxin B, licomycin, chloramphenicol, but resistant to tetracycline. Detailed phenotypic characteristics were listed in the species description. The complete morphological and physiological differences between the species are presented in Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003eChemotaxonomic characteristics\u003c/h2\u003e \u003cp\u003eThe major fatty acids (\u0026gt;\u0026thinsp;10.0% of the total fatty acids) of strain HH7-29\u003csup\u003eT\u003c/sup\u003e were iso-C\u003csub\u003e15:0\u003c/sub\u003e (35.0%) and anteiso-C\u003csub\u003e15:0\u003c/sub\u003e (15.4%). This profile is similar to that of the related species (except for \u003cem\u003eJ. campisalis\u003c/em\u003e), but with a rather wide range of abundance (11.0\u0026ndash;57.1% and 15.2\u0026ndash;53.5%, respectively). Furthermore, strain HH7-29\u003csup\u003eT\u003c/sup\u003e differs from the closely related species in terms of the proportions of certain moderate fatty acid components. For example, strain HH7-29\u003csup\u003eT\u003c/sup\u003e had significantly lower levels of alcohol-C\u003csub\u003e17:0\u003c/sub\u003e (6.0%) compared with reference strain \u003cem\u003eJ. salarius\u003c/em\u003e KCTC 13257\u003csup\u003eT\u003c/sup\u003e (11.5%) (Table S7). The polar lipids compositions of strain HH7-29\u003csup\u003eT\u003c/sup\u003e was identified as phosphatidylglycerol (PG), diphosphatidylglycerol (DPG) and two unidentified phospholipids (PL1 and PL2) (Fig. S7). Therein, PG, DPG and PL2 were detected in \u003cem\u003eJ. salarius\u003c/em\u003e KCTC 13257\u003csup\u003eT\u003c/sup\u003e, but no PL1 (Fig. S7). Furthermore, the main respiratory quinones of strain HH7-29\u003csup\u003eT\u003c/sup\u003e were MK-7 and MK-8, in line with all other members of the \u003cem\u003eJeotgalibacillus\u003c/em\u003e representatives.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003eTAXONMIC CONCLUSION\u003c/h2\u003e \u003cp\u003eThe phenotypic, phylogenetic, genomic, physiological and chemotaxonomic features all support the hypothesis that strain HH7-29\u003csup\u003eT\u003c/sup\u003e represents a novel species of the genus \u003cem\u003eJeotgalibacillus\u003c/em\u003e, for which the name \u003cem\u003eJeotgalibacillus haloalkaliphilus\u003c/em\u003e sp. nov. is proposed here.\u003c/p\u003e \u003cp\u003e \u003cb\u003eDESCRIPTION OF\u003c/b\u003e \u003cb\u003eJEOTGALIBACILLUS HALOALKALIPHILUS\u003c/b\u003e \u003cb\u003eSP. NOV.\u003c/b\u003e\u003c/p\u003e \u003cp\u003e \u003cem\u003eJeotgalibacillus haloalkaliphilus\u003c/em\u003e [hal.o.al.ka.li\u0026rsquo;phi.lus. Gr. masc. n. hals, salt; N.L. neut. n. alkali, alkali (indeclinable); Gr. masc. adj. philos, loving; N.L. masc. adj. haloalkaliphilus, loving briny and alkaline media]\u003c/p\u003e \u003cp\u003eCells are Gram-stain positive, aerobic and rod-shaped (0.6\u0026ndash;0.9\u0026times;1.0\u0026ndash;1.6 \u0026micro;m) with polar flagella. Colonies are yellow, circular, raised and approximately 2 mm in diameter on TYS agar after incubation for 2 days. Growth occurs at 15\u0026ndash;40℃ (optimum, 32℃), in 0.5\u0026ndash;24% NaCl (optimum, 2\u0026ndash;9%) and at pH 6.0\u0026ndash;12.0 (optimum, pH 8.0\u0026ndash;8.5). The strain is catalase-positive and oxidase-negative. Casein, milk, Tweens 20, 40, 60 and 80 are hydrolysed, but starch, gelatin, cellulose and sodium alginate are not hydrolysed. In API 20NE tests, positive for nitrate reduction, urease, arginine dihydrolase, hydrolysis of aesculin, acid production from \u003cem\u003eβ\u003c/em\u003e-galactosidase and assimilation of maltose, gluconate, malic acid and citric acid, but negative for indole production, hydrolysis of gelatin, acidification of glucose and assimilation of glucose, arabinose, mannose, mannitol, N-acetylglucosamine, capric acid, adipate and phenylacetic acid. In API ZYM tests, there are positive reactions for alkaline phosphatase, esterase lipase (C8), trypsin, and \u003cem\u003eα\u003c/em\u003e-chymotrypsin, but negative reactions for esterase (C4), lipase (C14), leucine arylamidase, valine aminopeptidase, cystine aminopeptidase, acid phosphatase, naphthol-AS-BI-phosphohydrolase, \u003cem\u003eα\u003c/em\u003e-galactosidase, \u003cem\u003eβ\u003c/em\u003e-galactocosidase, \u003cem\u003eβ\u003c/em\u003e-glucuronidase, \u003cem\u003eα\u003c/em\u003e-glucosidase, \u003cem\u003eβ\u003c/em\u003e-glucosinase, \u003cem\u003eα\u003c/em\u003e-mannosidase, \u003cem\u003eβ\u003c/em\u003e-fucosidase, and N-acetyl-\u003cem\u003eβ\u003c/em\u003e-glucosaminidase. The predominant fatty acids are iso-C\u003csub\u003e15:0\u003c/sub\u003e and anteiso-C\u003csub\u003e15:0\u003c/sub\u003e. The polar lipids consist of phosphatidylglycerol, diphosphatidylglycerol, and two unidentified phospholipids. The major menaquinones are MK-7 and MK-8.\u003c/p\u003e \u003cp\u003eThe type strain, HH7-29\u003csup\u003eT\u003c/sup\u003e (=\u0026thinsp;KCTC 43417\u003csup\u003eT\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;MCCC 1K07541\u003csup\u003eT\u003c/sup\u003e), was isolated from the estuary of the Fenhe River into the Yellow River of Shanxi, China. The genomic DNA G\u0026thinsp;+\u0026thinsp;C content of the type strain is 43.0%. The NCBI GenBank accession numbers for the 16S rRNA gene sequence and the draft genome sequence are OR856029 and JAXQNN000000000, respectively.\u003c/p\u003e \u003c/div\u003e"},{"header":"Abbreviations","content":"\u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd width=\"26.036866359447004%\"\u003e\n \u003cp\u003eMCCC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"73.963133640553%\"\u003e\n \u003cp\u003eMarine culture collection of China\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"26.036866359447004%\"\u003e\n \u003cp\u003eKCTC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"73.963133640553%\"\u003e\n \u003cp\u003eKorean collection for type cultures\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"26.036866359447004%\"\u003e\n \u003cp\u003eMEGA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"73.963133640553%\"\u003e\n \u003cp\u003eMolecular evolutionary genetics analysis\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"26.036866359447004%\"\u003e\n \u003cp\u003eOCs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"73.963133640553%\"\u003e\n \u003cp\u003eOrthologous clusters\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"26.036866359447004%\"\u003e\n \u003cp\u003eANI\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"73.963133640553%\"\u003e\n \u003cp\u003eAverage nucleotide identity\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"26.036866359447004%\"\u003e\n \u003cp\u003edDDH\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"73.963133640553%\"\u003e\n \u003cp\u003eDigital DNA-DNA hybridization\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"26.036866359447004%\"\u003e\n \u003cp\u003eAAI\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"73.963133640553%\"\u003e\n \u003cp\u003eAmino acid identity\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"26.036866359447004%\"\u003e\n \u003cp\u003ePOCP\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"73.963133640553%\"\u003e\n \u003cp\u003ePercentage of conserved proteins\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"26.036866359447004%\"\u003e\n \u003cp\u003eTLC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"73.963133640553%\"\u003e\n \u003cp\u003eThin-layer chromatography\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e"},{"header":"Declarations","content":"\u003cp\u003eThe GenBank accession numbers for the 16S rRNA gene sequence and the genome sequence of strain HH7-29\u003csup\u003eT\u003c/sup\u003e are\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eOR856029 and JAXQNN000000000, respectively.\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eAuthor contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eYFZ and YLP designed the experiments and wrote the manuscript. YHX, BY and MYC performed the genome analysis. ZLZ and LQH revised the manuscript. YL supervised the experiments.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis work was funded by\u0026nbsp;the National Natural Science Foundation of China (32300115),\u0026nbsp;the Basic Research Program of Shanxi Province (20210302124004), by the Scientific and Technological Innovation Programs of Shanxi Agricultural University (2020BQ39), by the Scientific and Technological Innovation Programs of Higher Education Institutions in Shanxi Province (2021L134) and by the Excellent Talents Come to Shanxi to Reward Scientific Research Projects (SXYBKY2019025)..\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData availability\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll data obtained in this study are shown within the manuscript and supplemental materials.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors have no relevant financial or non-financial interests to disclose.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics approval\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors have declared that no ethical issues exist.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAziz RK, Bartels D, Best AA, DeJongh M, Disz T et al (2008) The RAST Server: rapid annotations using subsystems technology. BMC genomics 9:75\u003c/li\u003e\n\u003cli\u003eBurg MB, Ferraris JD (2008) Intracellular organic osmolytes: function and regulation. J Biol Chem 283:7309\u0026ndash;7313\u003c/li\u003e\n\u003cli\u003eChen YG, Peng DJ, Chen QH, Zhang YQ, Tang SK et al (2010) \u003cem\u003eJeotgalibacillus soli \u003c/em\u003esp. nov., isolated from non-saline forest soil, and emended description of the genus \u003cem\u003eJeotgalibacillus\u003c/em\u003e. Antonie van Leeuwenhoek 98(3):415\u0026ndash;421\u003c/li\u003e\n\u003cli\u003eChun J, Oren A, Ventosa A, Christensen H, Arahal DR et al (2018) Proposed minimal standards for the use of genome data for the taxonomy of prokaryotes. Int J Syst Evol Microbiol 68(1):461\u0026ndash;466\u003c/li\u003e\n\u003cli\u003eCollins MD, Jones D (1980) Lipids in the classification and identification of coryneform bacteria containing peptidoglycans based on 2, 4-diaminobutyric acid. Appl Bacteriol 48:459\u0026ndash;470\u003c/li\u003e\n\u003cli\u003eCunha S, Tiago I, Paiva G, Nobre F, da Costa MS et al (2012) \u003cem\u003eJeotgalibacillus soli\u003c/em\u003e sp. nov., a Gram-stain-positive bacterium isolated from soil. Int J Syst Evol Microbiol 62(Pt 3):608\u0026ndash;612\u003c/li\u003e\n\u003cli\u003eEdgar RC (2004) MUSCLE: Multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Res 32:1792-1797\u003c/li\u003e\n\u003cli\u003eFelsenstein J (1981) Evolutionary trees from DNA sequences: a maximum likelihood approach. J Mol Evol 17:368\u0026ndash;376\u003c/li\u003e\n\u003cli\u003eIto M, Xu H, Guffanti AA, Wei Y, Zvi L et al (2004) The voltage-gated Na\u003csup\u003e+\u003c/sup\u003e channel NaVBP has a role in motility, chemotaxis, and pH homeostasis of an alkaliphilic Bacillus. Proc Natl Acad Sci 101(29):10566\u0026ndash;10571\u003c/li\u003e\n\u003cli\u003eJackman SD, Vandervalk BP, Mohamadi H, Chu J, Yeo S et al (2017) ABySS 2.0: resource-efficient assembly of large genomes using a Bloom filter. Genome Res 27:768\u0026ndash;777\u003c/li\u003e\n\u003cli\u003eJiang HN, Yun ST, Wang BX, Zhang MJ, Ma Y et al (2022) \u003cem\u003eJeotgalibacillus aurantiacus\u003c/em\u003e sp. nov., a novel orange-pigmented species with a carotenoid biosynthetic gene cluster, isolated from wetland soil. Antonie van Leeuwenhoek 115(6):773\u0026ndash;782\u003c/li\u003e\n\u003cli\u003eJones DM (1981) Manual of Methods for General Bacteriology. J Clin Pathol 34(9):1069\u003c/li\u003e\n\u003cli\u003eJones DT, Taylor WR, Thornton JM (1992) The rapid generation of mutation data matrices from protein sequences. Comput Appl Biosci 8:275\u0026ndash;282 \u003c/li\u003e\n\u003cli\u003eKim M, Oh HS, Park SC, Chun J (2014) Towards a taxonomic coherence between average nucleotide identity and 16S rRNA gene sequence similarity for species demarcation of prokaryotes. Int J Syst Evol Microbiol 64(Pt 2):346\u0026ndash;351\u003c/li\u003e\n\u003cli\u003eKomagata K, Suzuki KI (1987) Lipid and cell wall analysis in bacterial systematics. Methods Microbiol 19:161-207\u003c/li\u003e\n\u003cli\u003eKoichiro T, Glen S, Sudhir K (2021) MEGA11: molecular evolutionary genetics analysis version 11. Mol Biol Evol 38:3022\u0026ndash;3027\u003c/li\u003e\n\u003cli\u003eKrulwich TA, Hicks DB, Ito M (2009) Cation/proton antiporter complements of bacteria: why so large and diverse? Mol Microbiol 74(2):257\u0026ndash;260\u003c/li\u003e\n\u003cli\u003eLechner M, Findeiss S, Steiner L, Marz M, Stadler PF et al (2011) Proteinortho: Detection of (Co-)orthologs in large-scale analysis. BMC bioinformatics 12:124\u003c/li\u003e\n\u003cli\u003eLiew KJ, Lim L, Woo HY, Chan KG, Shamsir MS et al (2018) Purification and characterization of a novel GH1 beta-glucosidase from \u003cem\u003eJeotgalibacillus malaysiensis\u003c/em\u003e. Int J Syst Evol Microbiol 115:1094\u0026ndash;1102\u003c/li\u003e\n\u003cli\u003eLi Y, Zhang Z, Xu Z, Fang D, Wang ET et al (2018) \u003cem\u003eJeotgalibacillus proteolyticus\u003c/em\u003e sp. nov., a protease-producing bacterium isolated from ocean sediments. Int J Syst Evol Microbiol 68(12):3790\u0026ndash;3795\u003c/li\u003e\n\u003cli\u003eMedema MH, Blin K, Cimermancic P, de Jager V, Zakrzewski P et al (2011) antiSMASH: rapid identification, annotation and analysis of secondary metabolite biosynthesis gene clusters in bacterial and fungal genome sequences. Nucleic Acids Res 39:W339\u0026ndash;W346\u003c/li\u003e\n\u003cli\u003eMeier-Kolthoff JP, Auch AF, Klenk HP, G\u0026ouml;ker M （2013） Genome sequence-based species delimitation with confidence intervals and improved distance functions. BMC Bioinformatics 14:60\u003c/li\u003e\n\u003cli\u003eMongodin EF, Nelson KE, Daugherty S, Deboy RT, Wister J et al (2005) The genome of \u003cem\u003eSalinibacter ruber\u003c/em\u003e: convergence and gene exchange among hyperhalophilic bacteria and archaea. Proc Natl Acad Sci USA 102:18147\u0026ndash;18152\u003c/li\u003e\n\u003cli\u003ePardi F, Guillemot S, Gascuel O (2010) Robustness of phylogenetic inference based on minimum evolution. Bull Math Biol 72:1820\u0026ndash;1839\u003c/li\u003e\n\u003cli\u003eQin QL, Xie BB, Zhang XY, Chen XL, Zhou BC et al (2014) A proposed genus boundary for the prokaryotes based on genomic insights. J Bacteriol 196(12):2210\u0026ndash;2215\u003c/li\u003e\n\u003cli\u003eRüger HJ, Richter G (1979) \u003cem\u003eBacillus globisporus\u003c/em\u003e subsp. \u003cem\u003emarinus\u003c/em\u003e subsp. nov. Int J Syst Evol Microbiol 29:196-203\u003c/li\u003e\n\u003cli\u003eSaitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406\u0026ndash;425\u003c/li\u003e\n\u003cli\u003eSmibert RM, Krieg NR (1981) Manual of methods for general bacteriology. American Society for Microbiology, Washington, DC, pp 409\u0026ndash;443\u003c/li\u003e\n\u003cli\u003eSoliev AB, Hosokawa K, Enomoto K (2011) Bioactive pigments from marine bacteria: applications and physiological roles. Evid Based Complement Alternat Med 2011:1\u0026ndash;17\u003c/li\u003e\n\u003cli\u003eSrinivas A, Divyasree B, Sasikala C, Tushar L, Bharti D et al (2016) Description of \u003cem\u003eJeotgalibacillus alkaliphilus\u003c/em\u003e sp. nov., isolated from a solar salt pan, and \u003cem\u003eJeotgalibacillus terrae\u003c/em\u003e sp. nov., a name to replace \u0026apos;\u003cem\u003eJeotgalibacillus soli\u003c/em\u003e\u0026apos; Chen \u003cem\u003eet al\u003c/em\u003e. 2010. Int J Syst Evol Microbiol 66(12):5167\u0026ndash;5172\u003c/li\u003e\n\u003cli\u003eTatusova T, DiCuccio M, Badretdin A, Chetvernin V, Nawrocki EP et al (2016) NCBI prokaryotic genome annotation pipeline. Nucleic Acids Res 44:6614\u0026ndash;6624\u003c/li\u003e\n\u003cli\u003eThompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673\u0026ndash;4680\u003c/li\u003e\n\u003cli\u003eVargas C, Argando\u0026ntilde;a M, ReinaBueno M, Rodr\u0026iacute;guez-Moya J, Fern\u0026aacute;ndez-Auni\u0026oacute;n C et al (2008) Unravelling the adaptation responses to osmotic and temperature stress in \u003cem\u003eChromohalobacter salexigens\u003c/em\u003e, a bacterium with broad salinity tolerance. Saline Syst 4:14\u003c/li\u003e\n\u003cli\u003eWargo MJ (2013) Homeostasis and catabolism of choline and glycine betaine: lessons from Pseudomonas aeruginosa. Appl Environ Microbiol 79:2112\u0026ndash;2120\u003c/li\u003e\n\u003cli\u003eWeisburg WG, Barns SM, Pelletier DA, Lane DJ (1991) 16S ribosomal DNA amplification for phylogenetic study. J Bacteriol 173:697\u0026ndash;703\u003c/li\u003e\n\u003cli\u003eYaakop AS, Chan KG, Ee R, Kahar UM, Kon WC et al (2015) Isolation of \u003cem\u003eJeotgalibacillus malaysiensis\u003c/em\u003e sp. nov. from a sandy beach, and emended description of the genus \u003cem\u003eJeotgalibacillus\u003c/em\u003e. Int J Syst Evol Microbiol 65(7):2215\u0026ndash;2221\u003c/li\u003e\n\u003cli\u003eYaakop AS, Chan KG, Gan HM, Goh KM (2015) Draft genome of \u003cem\u003eJeotgalibacillus campisalis\u003c/em\u003e SF-57(T), a moderate halophilic bacterium isolated from marine saltern. Marine genomics 23:59\u0026ndash;60\u003c/li\u003e\n\u003cli\u003eYoon JH, Kang SJ, Schumann P, Oh TK (2010) \u003cem\u003eJeotgalibacillus salarius\u003c/em\u003e sp. nov., isolated from a marine saltern, and reclassification of \u003cem\u003eMarinibacillus marinus\u003c/em\u003e and \u003cem\u003eMarinibacillus campisalis\u003c/em\u003e as \u003cem\u003eJeotgalibacillus marinus\u003c/em\u003e comb. nov. and \u003cem\u003eJeotgalibacillus campisalis\u003c/em\u003e comb. nov., respectively. Int J Syst Evol Microbiol 60(Pt 1):15\u0026ndash;20\u003c/li\u003e\n\u003cli\u003eYoon JH, Kim IG, Schumann P, Oh TK, Park YH (2004) \u003cem\u003eMarinibacillus campisalis\u003c/em\u003e sp. nov., a moderate halophile isolated from a marine solar saltern in Korea, with emended description of the genus \u003cem\u003eMarinibacillus\u003c/em\u003e. Int J Syst Evol Microbiol 54(Pt 4):1317\u0026ndash;1321\u003c/li\u003e\n\u003cli\u003eYoon JH, Weiss N, Lee KC, Lee IS, Kang KH et al (2001) \u003cem\u003eJeotgalibacillus alimentarius\u003c/em\u003e gen. nov., sp. nov., a novel bacterium isolated from jeotgal with L-lysine in the cell wall, and reclassification of \u003cem\u003eBacillus marinus\u003c/em\u003e R\u0026uuml;ger 1983. as \u003cem\u003emMrinibacillus marinus\u003c/em\u003e gen nov., comb. nov. Int J Syst Evol Microbiol 51(Pt 6):2087\u0026ndash;2093\u003c/li\u003e\n\u003cli\u003eYoon SH, Ha SM, Lim JM, Kwon SJ, Chun J (2017) A large-scale evaluation of algorithms to calculate average nucleotide identity. Antonie van Leeuwenhoek. Int J Syst Evol Microbiol 110:1281\u0026ndash;1286\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"antonie-van-leeuwenhoek","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"anto","sideBox":"Learn more about [Antonie van Leeuwenhoek](https://www.springer.com/journal/10482)","snPcode":"10482","submissionUrl":"https://submission.nature.com/new-submission/10482/3","title":"Antonie van Leeuwenhoek","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Jeotgalibacillus haloalkaliphilus, polyphasic taxonomy, estuary, alkaliphilic and halotolerant","lastPublishedDoi":"10.21203/rs.3.rs-3880322/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-3880322/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eA Gram-stain positive, aerobic, alkaliphilic and halotolerant bacterium, designated HH7-29\u003csup\u003eT\u003c/sup\u003e, was isolated from the estuary of the Fenhe River into the Yellow River in Shanxi Province, PR China. Growth occurred at pH 6.0\u0026ndash;12.0 (optimum, pH 8.0\u0026ndash;8.5) and 15\u0026ndash;40℃ (optimum, 32℃) with 0.5\u0026ndash;24% NaCl (optimum, 2\u0026ndash;9%). The predominant fatty acids (\u0026gt;\u0026thinsp;10.0%) were iso-C\u003csub\u003e15:0\u003c/sub\u003e and anteiso-C\u003csub\u003e15:0\u003c/sub\u003e. The major menaquinones were MK-7 and MK-8. The polar lipids were phosphatidylglycerol, diphosphatidylglycerol and two unidentified phospholipids. Phylogenetic analyses based on the 16S rRNA gene sequence revealed that strain HH7-29\u003csup\u003eT\u003c/sup\u003e was a member of the genus \u003cem\u003eJeotgalibacillus\u003c/em\u003e, exhibiting high sequence similarity to the 16S rRNA gene sequences of \u003cem\u003eJeotgalibacillus alkaliphilus\u003c/em\u003e JC303\u003csup\u003eT\u003c/sup\u003e (98.4%), \u003cem\u003eJeotgalibacillus salarius\u003c/em\u003e ASL-1\u003csup\u003eT\u003c/sup\u003e (98.1%) and \u003cem\u003eJeotgalibacillus alimentarius\u003c/em\u003e YKJ-13\u003csup\u003eT\u003c/sup\u003e (98.1%). The genomic DNA G\u0026thinsp;+\u0026thinsp;C content was 43.0%. Gene annotation showed that strain HH7-29\u003csup\u003eT\u003c/sup\u003e had lower protein isoelectric points (pIs) and possessed genes related to ion transport and organic osmoprotectant uptake, implying its potential tolerance to salt and alkali. The average nucleotide identity, digital DNA\u0026ndash;DNA hybridization values, amino acid identity values, and percentage of conserved proteins values between strain HH7-29\u003csup\u003eT\u003c/sup\u003e and its related species were 71.1\u0026ndash;83.8%, 19.5\u0026ndash;27.4%, 66.5\u0026ndash;88.4% and 59.8\u0026ndash;76.6%, respectively. Based on the analyses of phenotypic, chemotaxonomic, phylogenetic and genomic features, strain HH7-29\u003csup\u003eT\u003c/sup\u003e represents a novel species of the genus \u003cem\u003eJeotgalibacillus\u003c/em\u003e, for which the name \u003cem\u003eJeotgalibacillus haloalkaliphilus\u003c/em\u003e sp. nov. is proposed. The type strain is HH7-29\u003csup\u003eT\u003c/sup\u003e (=\u0026thinsp;KCTC 43417\u003csup\u003eT\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;MCCC 1K07541\u003csup\u003eT\u003c/sup\u003e).\u003c/p\u003e","manuscriptTitle":"Jeotgalibacillus haloalkaliphilus sp. nov., a novel alkaliphilic and halotolerant bacterium, isolated from the estuary of the Fenhe River into the Yellow River","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-01-23 13:32:44","doi":"10.21203/rs.3.rs-3880322/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2024-01-22T15:02:13+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-01-22T14:58:44+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2024-01-20T07:01:09+00:00","index":"","fulltext":""},{"type":"submitted","content":"Antonie van Leeuwenhoek","date":"2024-01-20T01:49:01+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"antonie-van-leeuwenhoek","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"anto","sideBox":"Learn more about [Antonie van Leeuwenhoek](https://www.springer.com/journal/10482)","snPcode":"10482","submissionUrl":"https://submission.nature.com/new-submission/10482/3","title":"Antonie van Leeuwenhoek","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"830ff997-9ff9-4d57-b3ea-8aca6fb4bd17","owner":[],"postedDate":"January 23rd, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2024-05-07T20:04:48+00:00","versionOfRecord":{"articleIdentity":"rs-3880322","link":"https://doi.org/10.1007/s10482-024-01968-8","journal":{"identity":"antonie-van-leeuwenhoek","isVorOnly":false,"title":"Antonie van Leeuwenhoek"},"publishedOn":"2024-05-01 19:58:30","publishedOnDateReadable":"May 1st, 2024"},"versionCreatedAt":"2024-01-23 13:32:44","video":"","vorDoi":"10.1007/s10482-024-01968-8","vorDoiUrl":"https://doi.org/10.1007/s10482-024-01968-8","workflowStages":[]},"version":"v1","identity":"rs-3880322","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-3880322","identity":"rs-3880322","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}