Description of Prasinibacter corallicola gen. nov., sp. nov., a zeaxanthin-producing bacterium isolated from stony coral Porites lutea

Description of Prasinibacter corallicola gen. nov., sp. nov., a zeaxanthin-producing bacterium isolated from stony coral Porites lutea | 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 Description of Prasinibacter corallicola gen. nov., sp. nov., a zeaxanthin-producing bacterium isolated from stony coral Porites lutea Kefu Yu, Guanghua Wang, Jianfeng Liu, Yuanjin Li, Jin Li, Jixin Luo, and 4 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-1105606/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 4 You are reading this latest preprint version Abstract Thermal stress is considered one of the main causes of mass scleractinian coral degradation; however, it is still unknown how corals can adapt to future global warming. In this study, ten genera of coral-associated flavobacteria were shown to produce zeaxanthin, a carotenoid antioxidant, which may help coral holobionts to alleviate thermal stress. In addition, a novel zeaxanthin-producing flavobacterium, designated R38 T , was identified using polyphasic taxonomy. Although strain R38 T shared a maximum 16S rRNA gene sequence similarity of 93% with Mesoflavibacter aestuarii KYW614 T , phylogenetic analyses based on whole genome and 16S rRNA gene sequences revealed that strain R38 T forms a distinct branch in a robust cluster composed of strain R38 T and Leptobacterium flavescens KCTC 22160 T under the family Flavobacteriaceae . Strain R38 T exhibited average nucleotide identities of 70.2% and 72.5% for M. aestuarii KYW614 T and L. flavescens KCTC 22160 T , respectively. The only detected respiratory quinone was menaquinone 6 (MK-6). The genomic DNA G + C content was 33.2 mol%. The major polar lipids were phosphatidylmethylethanolamine, phosphatidylethanolamine, one unidentified ninhydrin phospholipid, three unidentified ninhydrin-positive lipids, and three unidentified lipids. The major cellular fatty acids were iso-C 15 : 0, iso-C 15 : 0 ω6c, C 16:2 DMA, and C 13:1 ω3c. The distinct biochemical, chemotaxonomic, phylogenetic, and phylogenomic differences from validly published taxa suggest that strain R38 T represents a new species of a new genus, for which Prasinibacter corallicola gen. nov., sp. nov. is proposed. The type strain R38 T (= MCCC 1K03889 T = KCTC 72444 T ). Prasinibacter corallicola polyphasic taxonomy 16S rRNA gene zeaxanthin Figures Figure 1 Figure 2 Introduction Global coral reefs have degraded dramatically in recent decades (Hughes et al. 2003 ; Bellwood et al. 2004 ) and massive corals have replaced branched corals as the dominant assemblages (Perry et al. 2015 ; Yu et al. 2019 ). Thermal stress is one of the main causes of scleractinian coral degradation (Gardner et al. 2003 ; Hughes et al. 2017 ; Hughes et al. 2019 ). Recently, it was reported that the phycosphere bacteria Muricauda sp. GF1 protects coral endosymbionts from thermal stress by producing zeaxanthin (Motone et al. 2020 ). Zeaxanthin is usually synthesized in flavobacteria from terpenoids by the combinations of phytoene synthase (CrtB), phytoene dehydrogenase (CrtI), lycopene cyclase (CrtY), and β-carotene hydroxylase (CrtZ) (Zhang et al. 2018 ). According to the List of Prokaryotic names with Standing in Nomenclature ( https://lpsn.dsmz.de/ ), to date, up to 151 validly published genera have been assigned to the family Flavobacteriaceae . Genomic and physiological analyses have indicated that flavobacteria can degrade a diverse range of carbohydrates and proteins (Bauer et al. 2006 ; Qin et al. 2010 ; Gavriilidou et al. 2020 ), and a few are animal pathogens (Duchaud et al. 2007 ; Loch and Faisal 2015 ; Adamek et al. 2018 ). To determine the roles of flavobacteria in coral health, pure cultures were isolated from thermal tolerant corals: Porites lutea , Galaxea fascicularis , and Favia sp. The taxonomic position of a new coral-associated flavobacterium, strain R38 T , was identified using polyphasic identification, and the results indicated that strain R38 T and other coral-associated flavobacteria can also produce zeaxanthin. Therefore, coral-associated flavobacteria may help alleviate host thermal stress caused by global warming. Materials And Methods Isolation, cultivation, and maintenance Massive coral P. lutea was collected from Weizhou Island (109º 08′ 35′′ E, 21º 03′ 42′′ N), in the Beibu Gulf, China. G. fascicularis and Favia sp . were collected from Daya Bay (114º 38′ 32 ′′E, 22º 34′ 32′′ N), China. The coral pieces (approximately 1 cm × 1 cm) were washed twice using sterile natural seawater, then the coral tissue was homogenized using silica beads on vortex mixer after scraping with scissors. The homogenate was diluted by ten-fold using sterile seawater, and 100 μl of each dilution was spread on modified R2A plate (marine R2A plate, R2A agar was obtained from BD, which was dissolved in natural seawater, pH7.6). Bacteria were incubated at 25 °C for two weeks. Colonies were picked and purified on marine R2A plates, and stored at -70 °C in R2A broth (Haibo, China)/glycerol (4:1, v/v). Preliminary identification using 16S rRNA gene sequence indicated that strain R38 T (from P. lutea ) may represent a new species in a new genus, therefore, polyphasic identification was performed to identify its exact taxonomic position. Type strains Leptobacterium flavescens KCTC 22160 T and Spongiivirga citrea KCTC 32990 T , obtained from the Korean Collection for Type Cultures (KCTC), were used as references. Both the new isolate and the reference type strains grew well on marine agar 2216 (BD). Physiological and biochemical analyses Cellular morphology was observed using an optical microscope (Olympus BX53) and transmission electron microscope (Tecnai G2 F30 S-TWIN/X-MAX 80) after incubation for 2-3 days in marine broth 2216 (BD) at 30 °C. Cell mobility was tested using the hanging drop technique (Bernardet et al. 2002). The Gram reaction was determined as described by Gerhardt et al. (1994). Catalase activity was determined by observing bubble production in a 3 % (v/v) hydrogen peroxide solution, and oxidase activity was determined using oxidase test strips (Huankai, China). The ability to form endospores was examined as described by Dong & Cai (2001). NaCl requirement and tolerance were tested at 30 °C for 7 days in R2A liquid medium (Haibo, China) with NaCl concentrations ranging from 0-16% (w/v), namely 0, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, and 16 % (w/v). Growth at different pH, adjusted with different buffers (1 interval, pH 4-5, 0.1 M citrate; pH 6-8, 0.1 M phosphate; pH 9-11, 0.1 M carbonate), was tested in R2A liquid medium [including 1% NaCl (w/v)] at 30 °C for 7 days. The optimal growth temperature for bacteria on marine agar 2216 (BD) plates was determined after 7-30 days of growth at temperatures of 4, 10, 15, 20, 25, 30, 33, 37, and 40 °C. Metabolism ability was characterized using API 20NE strips, API ZYM strips, and Biolog Gen III microplates according to the manufacturer’s protocols. Anaerobic fermentation was determined using API 50CH strips according to the manufacturer’s protocol with the inoculum medium being marine broth 2216 (BD). Chemotaxonomic characterization The biomass of strain R38 T and the reference strain for cellular fatty acid analysis were acquired from the third quadrant of the streaked marine agar 2216 plate incubated at 28 °C. Cellular fatty acid composition was analyzed by gas chromatography (Agilent G6890N) and identified using the Sherlock Microbial Identification System (Version 6.0) according to the manufacturer’s instructions. Biomass for the analyses of quinones and polar lipids was obtained from marine broth 2216 after 3 days of incubation at 30 °C. Respiratory quinones were extracted as described by Collins (1994) and analyzed using reversed-phase high-performance liquid chromatography (HPLC) (Komagata & Suzuki, 1987). The isoprenoid quinones were eluted using a mixture of methanol/2-propanol (2:1, v/v) and flow rate of 1 ml/min at room temperature and detected by UV absorbance at 270 nm. Polar lipids were extracted as described by Kamekura (1993) and identified by spraying with ethanolic molybdophosphoric acid, molybdenum blue, ninhydrin, α-naphthol/sulfuric acid, and Dragendorff's reagent after two-dimensional thin layer chromatography (TLC) (Tindall 1990). Phylogenetic and phylogenomic analysis The 16S rRNA gene of strain R38 T was obtained using PCR amplification with the universal primers 27F and 1492R (Lane 1991) and sequenced using the Sanger method. The 16S rRNA gene sequence similarities were determined using the EzBioCloud (Yoon et al. 2017) and NCBI database. Alignment of 16S rRNA gene sequences was performed using the SINA software package (Pruesse et al. 2012) and the SILVA rRNA database. Phylogenetic trees were constructed using the maximum likelihood (Felsenstein 1981), neighbor-joining (Saitou & Nei 1987), and maximum-parsimony (Swofford 1993) algorithms in the software package MEGA version 7.0 (Kumar et al. 2016). The phylogenetic distance matrices were estimated using the Kimura two-parameter model (Kimura, 1980). The topology of the phylogenetic tree was analyzed using the bootstrap resampling method of Felsenstein (1985) with 1000 replicates. Whole-genome sequencing was performed using an Illumina HiSeq PE150 platform. Library construction was performed by PCR amplification of a 350 bp insert with A-tail ligated to paired-end adaptors, at the Beijing Novogene Bioinformatics Technology Co., Ltd. Good-quality paired reads were assembled into a number of scaffolds using the SOAPdenovo (Li et al. 2008; Li et al. 2010). Genomic information was extracted as described by Chun et al. (Chun et al. 2018). The phylogenetic tree was reconstructed using the up-to-date bacterial core gene set (UBCG v.3) according to the manual (Na et al. 2018). The average nucleotide identity (ANI) was calculated using the online ANI calculator (Yoon et al. 2017). The average amino acid identity (AAI) was calculated using the EzAAI (Kim et al. 2021). Carbohydrate-active enzyme annotation was performed using the dbCAN meta server (Zhang et al. 2018). Peptidases were annotated using the Hotpep-protease method (Bush, 2020) based on the Merops database. Zeaxanthin detection The production of zeaxanthin by bacteria from the family Flavobacteriaceae was assessed using both genome annotation and HPLC assay. The existence of coding genes for zeaxanthin biosynthesis enzymes such as phytoene desaturase, lycopene beta-cyclase, and beta-carotene 3-hydroxylase was checked using the Joint Genome Institute online server ( https://genome.jgi.doe.gov/portal/ ) and the NCBI genome server ( https://www.ncbi.nlm.nih.gov/genome/ ). Zeaxanthin was extracted from approximately 10 mg wet weight bacteria grown on an R2A plate using methanol and glass beads on a vortex for 30 seconds, and then analyzed using HPLC (Thermal Ultimate 3000). Samples were separated using an Agilent ZORBAX Eclipse XDB-C18 (250 mm, 5 µm particle size) at a column temperature of 35 °C, and the mobile phase comprised 90% (vol/vol) methanol containing 0.1% (v/v) formic acid at a flow rate of 1 ml/min (Motone et al. 2020). Zeaxanthin was checked using a photodiode array detector. Standard zeaxanthin was purchased from the Resource Platform of Standard Material (China). Results And Discussion Zeaxanthin production Coral bleaching caused by thermal stress is becoming increasingly serious (Hughes et al. 2018). In the last decade, coral bleaching has occurred at significantly higher sea surface temperatures (SSTs) (∼0.5 °C) than in the previous decade, suggesting that thermally susceptible genotypes may have declined and/or adapted (Sully et al. 2019). Zeaxanthin is a carotenoid antioxidant that has been shown to protect endosymbiotic Symbiodiniaceae algae, isolated from coral Galaxea fascicularis, from thermal and light stress (Motone et al. 2020). Coding genes for Zeaxanthin biosynthesis enzymes phytoene desaturase, lycopene beta-cyclase, and beta-carotene 3-hydroxylase were examined in 115 of the 177 total genera in Flavobacteriaceae , and over 50% of these genera (62) had all three enzymes (Table S1). Of note, all 11 strains of coral-associated flavobacteria (from approximately ten genera), including strain R38 T , were able to produce zeaxanthin according to HPLC analysis (Table 1). These results indicate that the Flavobacteriaceae family contains important zeaxanthin producers, and corals may benefit from these symbiotic flavobacteria when confronting thermal stress (Motone et al. 2020). However, zeaxanthin in coral ( Porites lutea and Acropora sp.) or Symbiodinium was below the limit of HPLC detection (Venn et al. 2006 and this study). Zeaxanthin is an intermediate product of the algal accessory photosynthetic pigments fucoxanthin and peridinin (Dautermann et al. 2020), and may be transformed immediately following synthesis in the coral holobiont. This is suggested by the detection of its precursor, β-carotene, and the downstream product, peridinin, in coral or Symbiodinium (Venn et al. 2006). Although zeaxanthin exchange between bacteria and the endosymbiont Symbiodinium has not been shown, similar effects of pure culture Muricauda sp. GF1 and zeaxanthin supplementation to cultured Symbiodiniaceae conducted by Motone et al. (2020) strongly supports this exchange. Therefore, flavobacterial zeaxanthin may support the biosynthesis of algal accessory photosynthetic pigments in the coral endosymbiont Symbiodinium . Finally, carotenoids can function as antioxidants via epoxidase/de-epoxidase reaction, regardless of the type of carotenoid in the final product (Krinsky 1989; Lacour et al. 2020). Morphological, physiological, and biochemical analyses Cells of bacterial strain R38 T were gram-negative, non-spore-forming, non-motile, aerobic rods. Cells were usually 0.3-0.5 μm wide and 0.9-2.0 μm long (Fig.1), being narrower than that of L. flavescens KCTC 22160 T , S. citrea KCTC 32990 T and Fulvibacter tottoriensis MTT-39 T , while being wider than that of Mesoflavibacter aestuarii KYW614 T (Table 2). Cells of strain R38 T could reduce nitrate to nitrogen, L. flavescens KCTC 22160 T and S. citrea KCTC 32990 T could only reduce nitrate to nitrite, whereas M. aestuarii KYW614 T (Lee et al. 2014) and F. tottoriensis MTT-39 T (Khan et al. 2008) could not reduce nitrate. Enzyme characterization of strain R38 T using API ZYM strips showed a spectrum similar to that of M. aestuarii KYW614 T and F. tottoriensis MTT-39 T with the absence of β-galactosidase, β-glucuronidase, α-glucosidase, β-glucosidase, N-acetyl-β-glucosaminidase, and α-mannosidase (Table 2) (Lee et al. 2014; Yoon et al. 2013). These enzyme results differed from those of L. flavescens KCTC 22160 T and S. citrea KCTC 32990 T (Table 2). Other characteristics of strain R38 T are listed in Table 2 and the species description. Chemotaxonomic characteristics The only respiratory quinone detected in strain R38 T was menaquinone 6 (MK-6), similar to L. flavescens KCTC 22160 T (Mitra et al. 2009), S. citrea KCTC 32990 T (Yoon et al. 2015), F. tottoriensis MTT-39 T (Khan et al. 2008), and Mesoflavibacter aestuarii KYW614 T (Lee et al. 2014). Strain R38 T contained iso-C15 : 0 (42.4 %), iso-C15 : 0ω6c (15.6 %), C 16:2 DMA (7.8 %), and C 13:1 ω3c (5.2 %) as the major cellular fatty acids, this profile was highly similar to that of L. flavescens KCTC 22160 T , although the proportion of individual components varied (Table S2). However, the individual fatty acid content of S. citrea KCTC 32990 T differed from that of strains R38 T and L. flavescens KCTC 22160 T (Table S2). The major fatty acids of strain R38 T , L. flavescens KCTC 22160 T , and S. citrea KCTC 32990 T were highly different from those of M. aestuarii KYW614 T and F. tottoriensis MTT-39 T (Table 2) (Lee et al. 2014; Yoon et al. 2013). The major polar lipids of strain R38 T were phosphatidyl-N-methylethanolamine, phosphatidylethanolamine, one unidentified ninhydrin phospholipid, three unidentified ninhydrin-positive lipids, and three unidentified lipids (Supplementary Fig.S1). This polar lipid profile was similar to that of L. flavescens KCTC 22160 T , but highly different from that of S. citrea KCTC 32990 T , which contains few types of polar lipids (Supplementary Fig.S1). However, strain R38 T could still be distinguished from L. flavescens KCTC 22160 T by its unidentified ninhydrin phospholipid (Supplementary Fig.S1). Furthermore, no phosphatidyl-N-methylethanolamine has been reported for M. aestuarii KYW614 T and F. tottoriensis MTT-39 T (Lee et al. 2014; Yoon et al. 2013). Therefore, the polar lipid profile distinguishes strain R38 T from any validly published taxon. Molecular characterization and phylogenetic analysis A nearly complete 16S rRNA gene sequence (1383 nt) of strain R38 T was obtained by Sanger sequencing and deposited in GenBank under accession number MN908337. Global alignment using the EzBioCloud database indicated that the most closely related neighbor of strain R38 T is Mesoflavibacter aestuarii KYW614 T , with a 16S rRNA gene similarity of 93%. The next most similar members were of Bizionia , Sabulilitoribacter , Gaetbulibacter , and Algibacter genera which showed 92.7-92.9% sequence similarity. However, 16S rRNA gene phylogenetic analysis based on the maximum-likelihood algorithm indicated that strain R38 T forms a distinct branch in a stable cluster composed of strain R38 T and L. flavescens KCTC 22160 T (Fig. 2). Neighbor-joining clustering also supports this tree topology (Supplementary Fig. S2). The close relationship between strain R38 T and L. flavescens KCTC 22160 T was also represented in the maximum-parsimony analysis, although a robust cluster was not formed (Supplementary Fig.S3). Genome properties and comparison The genome sequencing depth of strain R38 T was 333×, and the N50 was 750154 bp. A total of 11 contigs were obtained, the obtained genome size was 3.94 Mb, and the genomic DNA G+C content was 33.2 mol%. The genome sequencing depth of L. flavescens KCTC 22160 T was 154×, the N50 was 1032064 bp, a total of 9 contigs were obtained, the obtained genome size was 4.21 Mb, and the genomic DNA G+C content was 40.9 mol%. The genome sequencing depth of S. citrea KCTC 32990 T was 155×, the N50 was 637254 bp, a total of 17 contigs were obtained, the obtained genome size was 4.15 Mb, and the genomic DNA G+C content was 36.3 mol%. The genomes of closely related type strains were 3.05-5.03 MB, with G+C content of 33.2-55.3 mol% (Supplementary Table S3). Thus, strain R38 T has the lowest genomic G+C content (33.2 mol%). The complete 16S rRNA gene of strain R38 T obtained by genome sequencing was 1508 nt, and showed two nucleotide differences compared to the sequence obtained by Sanger sequencing. Meanwhile, the result obtained by Sanger sequencing indicated there were at least two copies of the 16S rRNA gene in cells of strain R38 T . The ANI of strain R38 T to L. flavescens KCTC 22160 T , S. citrea KCTC 32990 T , and M. aestuarii KYW614 T were 72.5%, 69.6%, and 70.2%, respectively. The AAI of strain R38 T to L. flavescens KCTC 22160 T was 74.6%, and for other type strains, the indices were lower than 70%. Similar to the 16S rRNA gene-based phylogenetic results, phylogenomic analysis based on 92 genes also indicated that strain R38 T forms a distinct branch in a stable cluster composed of strain R38 T and L. flavescens KCTC 22160 T (Supplementary Fig.S4). Approximately 37 families of carbohydrate-active enzymes were detected in strain R38 T , while in closely related type strains this number was between 39-100 (Supplementary Table S3). A limited quantity of glycoside hydrolases (16 vs 20-63) indicates that strain R38 T is weak in carbohydrate utilization (Supplementary Table S3). Approximately 71 families of peptidases were detected in strain R38 T . This quantity is higher than in most of the closely related type strains (Supplementary Table S3), indicating that strain R38 T is versatile in protein utilization. This carbohydrate and protein utilization pattern might have resulted from long-term bacteria-animal association. Taxonomic conclusion Based on phylogenetic analyses, strain R38 T was found to be associated with the family Flavobacteriaceae . The ANI of strain R38 T to closely related type strains (≤72.5%) indicates that strain R38 T belongs to a novel species (Chun et al. 2018), and both biochemical and chemotaxonomic characteristics (Table 2) support this species-level assignment. Furthermore, the low 16S rRNA gene similarities (≤93%) of strain R38 T to closely related type strains indicate that strain R38 T represents a new genus (Yarza et al. 2014), which is also supported by the differences in polar lipid profile (Table 2 & Supplementary Fig.S1). Therefore, strain R38 T represents a new species in a new genus under the family Flavobacteriaceae , for which Prasinibacter corallicola gen. nov., sp. nov. is proposed. Description of Prasinibacter gen. nov. Prasinibacter (Pra.si.ni.bac′ter. L. masc. adj. prasinus , yellowish-green; N.L. masc. bacter , rod: N.L. masc. n. Parisinibacter , a translucent yellowish green rod.) Cells are gram-negative, non-spore-forming, non-motile, aerobic rods. Catalase- and oxidase-positive. Nitrate is reduced to nitrogen. The only menaquinone is MK-6. The major polar lipids are phosphatidyl-N-methylethanolamine, phosphatidylethanolamine, one unidentified ninhydrin phospholipid, three unidentified ninhydrin-positive lipids, and three unidentified lipids. The type species is Prasinibacter corallicola . Member of the family Flavobacteriaceae . Description of Prasibacter corallicola sp. nov. Prasinibacter corallicola (co.ral.li.co′la. L. neut. n. corallum , coral; L. masc. suff. - cola , inhabitant dweller; N.L. n. corallicola , coral-dweller) The description is as for the genus with the following additional properties: Cells are usually 0.3-0.5 μm wide and 0.9-2.0 μm long. Colonies are yellow-green, circular, and smooth on marine agar 2216. Cells can grow at 15-33 °C (optimum 25-30°C), pH 5-10 (optimum 7-8) in 3-6% (w/v) NaCl (optimum 3-4%) in R2A liquid medium. Zeaxanthin is produced. Production of H 2 S does not occur. Starch is hydrolyzed. In the API 20NE test, nitrate reduction and protease are positive. In the API ZYM test, alkaline phosphatase, esterase (C4), esterase lipase (C8) and (C14), leucine arylamidase, valine arylamidase, cysteine arylamidase, trypsin, α-chymotrypsin, acid phosphatase, naphthol-AS-BI phosphohydrolase, and α-galactosidase are positive. In the Biolog Gen III microplate, dextrin, D-maltose, α-D-glucose, D-mannose, L-alanine, L-glutamic acid, L-histidine, α-keto-glutaric acid, L-malic acid, bromo-succinic acid, Tween 40, acetoacetic acid, and acetic acid are oxidized. The major fatty acids are iso-C15 : 0, iso-C15 : 0ω6c, C 16:2 DMA, and C 13:1 ω3c. The genomic DNA G+C ratio is 33.2 mol%. The type strain, R38 T (=MCCC 1K03889 T =KCTC 72444 T ) was isolated from stony coral Porites lutea collected from Weizhou Island in the Beibu Gulf, China. The GenBank accession number of the 16S rRNA gene sequence of the type strain is MN908337. Abbreviations KCTC The Korean Collection for Type Cultures MCCC The Marine Culture Collection of China ANI Average nucleotide identity Q Ubiquinone DPG Diphosphatidylglycerol PG Phosphatidylglycerol PME phosphatidylmethylethanolamine PE Phosphatidylethanolamine NL/AL Unidentified ninhydrin positive lipid NPL Unidentified ninhydrin positive phospholipid PL Unidentified phospholipid L Unidentified lipid Declarations Author contributions Guanghua Wang, Jianfeng Liu and Yuanjin Li isolated strains, performed experiments and wrote the manuscript. Biao Chen and Zhiheng Liao collected samples. Hongfei Su and Jiayuan Liang gave advices about bacteria cultivation. Kefu Yu conceived and designed the experiments and approved the final manuscript. Funding This research was supported by the National Natural Sciences Foundation of China (41866004, 42090041 and 42030502); National Key R & D Program of China (2018YFD0900803); Science and Technology Project of Guangxi (AD17129063, AA17204074 and AA18242026). Declarations Conflicts of interest We have no conflict of interest to declare. Ethical approval Not applicable. References Adamek M, Teitge F, Jung-Schroers V, Heling M, Gela D, Piackova V, Kocour M, Steinhagen D. (2018) Flavobacteria as secondary pathogens in carp suffering from koi sleepy disease. J Fish Dis 41(11):1631-1642. Bauer, M., Kube, M., Teeling, H., Richter, M., Lombardot, T., Allers, E., Würdemann, C.A., Quast, C., Kuhl, H., Knaust, F., Woebken, D., Bischof, K., Mussmann, M., Choudhuri, J.V., Meyer, F., Reinhardt, R., Amann, R.I. and Glöckner, F.O. (2006), Whole genome analysis of the marine Bacteroidetes ‘Gramella forsetii’ reveals adaptations to degradation of polymeric organic matter. 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Tables Table 1 Coral associated bacterial ability of zeaxanthin production R & B were from Porites lutea ; F was from Favia sp.; and K was from Galaxea fascicularis. *About 500 bp 16S rRNA gene sequence was used in the 27f end. +, positive; w, weak positive. Strains (accession No.) Taxonomic neighbor Identity (%) Zeaxanthin R38 T (MN908337) Mesoflavibacter aestuarii KYW614 T 93 + R33 T (MN908336) Poritiphilus flavus R33 T 100 + BMA10(MZ779023) Fulvivirga lutimaris TM-6 T 92.3 + BMA12(MZ779024) Fulvivirga lutimaris TM-6 T 90.5 w B25(MZ779016) Ulvibacter litoralis KMM 3912 T 94.9* + F16(MZ779017) Tenacibaculum skagerrakense D30 T 96.9* + F26(MZ779019) Aquimarina atlantica 22II-S11-z7 T 100* w F45(MZ779020) ‘Gramella jeungdoensis’ HMD3159 96.9* + F53(MZ779021) ‘Gaetbulibacter jejuensis’ CNURIC 14 100* + K6(MZ779018) Mesoflavibacter sabulilitoris GJMS-9 T 99.7* + K18(MZ779022) Tenacibaculum mesophilum NBRC 16307 T 95.1* + Table 2 Phenotypic characteristics of strain R38 T and related type strains Strains: 1, R38 T ; 2, Leptobacterium flavescens KCTC 22160 T ; 3, Spongiivirga citrea KCTC 32990 T ; 4, Mesoflavibacter aestuarii KYW614 T (data from Lee et al. 2014); 5, Fulvibacter tottoriensis MTT-39 T (data from Khan et al. 2008; Yoon et al. 2013). +, positive; -, negative; ND, unknown. Characteristics 1 2 3 4 † 5 ɸ Habitat Coral Marine sponge‡ Marine sponge† Seawater marine sediment Cell size (μm) 0.3-0.5×0.9-2.0 0.5-0.6×8.5-9.0‡ 0.4-0.5×2.0-3.0† 0.2×1.2-1.5 0.5-0.7×5-14 Colony color Yellow-green Pale-yellow Lemon-yellow Orange yellowish brown Temperature range (℃) 15-33 15-35 15-35 4-35 10-37 NaCl tolerance (%, w/v) 3-6 0.5-6 1-5 1-9 1-5 pH range 5-10 6-10 8-9 6-8 6-10 Nitrate reduction + + + - - API ZYM test β-galactosidase - + + - - β-glucuronidase - + + - - α-glucosidase - + + - - β-glucosidase - + + - - N-acetyl-b-glucosaminidase - + + - - α-mannosidase - + + - - Genome size (Mb) 3.94 4.21 4.15 2.92 ND ANI of strain R38 T to 100% 72.5% 69.6% 70.2% ND G+C mol% 33.2 40.9 36.3 32.1 35 Major polar lipid PME,PE,NLs,NPL,Ls PME,PE,NLs, Ls PME,NL,L PE, 2ALs, 3Ls PL,NLs,Ls Major cellular fatty acids iso-C15 : 0, iso-C15 : 0ω6c, C 16:2 DMA, C 13:1 ω3c iso-C15 : 0, iso-C15 : 0ω6c, C 16:2 DMA, C16 : 1ω6c iso-C15 : 0, iso-C15 : 0ω6c, C 16:2 DMA, C 15:0 C15 : 0, iso-C15 : 0, iso-C16 : 0 3-OH, iso-C17 : 0 3-OH, iso-C15 : 1 G, C16 : 1 ω7c/ω6c iso-C 15 : 1 ,C 16:0 3-OH, iso-C 17 : 0 3-OH, iso-C 15 :0 3-OH, iso-C 15 : 0 . ‡Data from Mitra et al. (2009); † Data from Yoon et al. (2015). Supplementary Files R38supplementmateria.docx SupplemetaryTableS3.xlsx Cite Share Download PDF Status: Under Review Version 1 posted Reviews received at journal 21 Mar, 2022 Reviewers invited by journal 19 Mar, 2022 Editor assigned by journal 03 Mar, 2022 First submitted to journal 02 Mar, 2022 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-1105606","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":92086639,"identity":"1b56d9c6-50f2-4808-9ebd-b3ebe7ddcf46","order_by":0,"name":"Kefu Yu","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAy0lEQVRIiWNgGAWjYBACCQbGB0DKBogZG4jVwmwApNJI13KYBIdJtjezSfO2nbfn7z/c+IChxo6BfzYB26R5DoO03E6ccSOx2YDhWDKDxJ0D+LXISeQfA2lJMJBgbJNgYDvAYCCRQECL/GOQLefsDfgPArX8I0KLtAQzSMsBxg0MiW1Ai4jQItmTzGw551wyxC+Jfck8EjcIaJE4fpjxxpsyO2CIHX/44MM3Ozn+GQS0gAATD4wFVMyDRyECMP4gStkoGAWjYBSMWAAADcY6dDoZ8VgAAAAASUVORK5CYII=","orcid":"https://orcid.org/0000-0003-3409-9945","institution":"South China Sea Institute of Oceanology Chinese Academy of Sciences","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Kefu","middleName":"","lastName":"Yu","suffix":""},{"id":92086630,"identity":"9717fb0e-c52c-4cc9-a1ed-da64288a3876","order_by":1,"name":"Guanghua Wang","email":"","orcid":"","institution":"South China Sea Institute of Oceanology Chinese Academy of Sciences","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Guanghua","middleName":"","lastName":"Wang","suffix":""},{"id":92086631,"identity":"e97eb50a-f219-45ac-b3a1-a920b00a830d","order_by":2,"name":"Jianfeng Liu","email":"","orcid":"","institution":"South China Sea Institute of Oceanology Chinese Academy of Sciences","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Jianfeng","middleName":"","lastName":"Liu","suffix":""},{"id":92086632,"identity":"baa90dc1-aa36-4cf6-b26d-8bf4a6e53211","order_by":3,"name":"Yuanjin Li","email":"","orcid":"","institution":"South China Sea Institute of Oceanology Chinese Academy of Sciences","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Yuanjin","middleName":"","lastName":"Li","suffix":""},{"id":92086633,"identity":"087a6c0a-15de-404c-b223-b14e1b1f4897","order_by":4,"name":"Jin Li","email":"","orcid":"","institution":"South China Sea Institute of Oceanology Chinese Academy of Sciences","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Jin","middleName":"","lastName":"Li","suffix":""},{"id":92086634,"identity":"f14e8ccb-48db-4c9d-8337-5a47abb48064","order_by":5,"name":"Jixin Luo","email":"","orcid":"","institution":"South China Sea Institute of Oceanology Chinese Academy of Sciences","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Jixin","middleName":"","lastName":"Luo","suffix":""},{"id":92086635,"identity":"c1709fbf-4d83-4c7e-ad4f-6989dadfbc2a","order_by":6,"name":"Biao Chen","email":"","orcid":"","institution":"South China Sea Institute of Oceanology Chinese Academy of Sciences","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Biao","middleName":"","lastName":"Chen","suffix":""},{"id":92086636,"identity":"a93b485d-3e2e-442c-9637-b68e351d7618","order_by":7,"name":"Zhiheng Liao","email":"","orcid":"","institution":"South China Sea Institute of Oceanology Chinese Academy of Sciences","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Zhiheng","middleName":"","lastName":"Liao","suffix":""},{"id":92086637,"identity":"9b0e3d41-c957-41eb-9f58-15f96fd67780","order_by":8,"name":"Hongfei Su","email":"","orcid":"","institution":"South China Sea Institute of Oceanology Chinese Academy of Sciences","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Hongfei","middleName":"","lastName":"Su","suffix":""},{"id":92086638,"identity":"2a7ee2c9-c792-4595-a608-73ef94e6cdda","order_by":9,"name":"Jiayuan Liang","email":"","orcid":"","institution":"South China Sea Institute of Oceanology Chinese Academy of Sciences","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Jiayuan","middleName":"","lastName":"Liang","suffix":""}],"badges":[],"createdAt":"2021-11-23 04:12:39","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-1105606/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-1105606/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":19496269,"identity":"4a7af44c-592b-441a-a3e9-0b803b9c925c","added_by":"auto","created_at":"2022-03-22 18:41:43","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":254901,"visible":true,"origin":"","legend":"\u003cp\u003eTransmission electronic microscopy of strain R38\u003csup\u003eT\u003c/sup\u003e. Bar, 0.5 µm.\u003c/p\u003e\u003cp\u003e\u003cbr\u003e\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-1105606/v1/1f0f599397eec42b52943979.png"},{"id":19496270,"identity":"36c08cd2-bfbb-404a-b73c-8ebbe4c0428c","added_by":"auto","created_at":"2022-03-22 18:41:43","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":38961,"visible":true,"origin":"","legend":"\u003cp\u003eMaximum likelihood phylogenetic tree based on the 16S rRNA gene sequences of strain R38\u003csup\u003eT\u003c/sup\u003e and related taxa. Sequences from the \u003cem\u003eEscherichia coli \u003c/em\u003ewas used as out group. Numbers at nodes indicate percentages of 1000 bootstrap resamplings; only values above 50% are shown. Bar, 0.05 substitutions per nucleotide position.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-1105606/v1/355316e2701f5e3c3e087046.png"},{"id":19496362,"identity":"68f90075-7089-4938-8749-08b9e02e2b67","added_by":"auto","created_at":"2022-03-22 18:44:47","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":663685,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-1105606/v1/9dcd8547-0b1b-49a7-a2ff-8f0df54bbd33.pdf"},{"id":19496361,"identity":"5f9685bd-b952-4366-99df-ac25c0959c34","added_by":"auto","created_at":"2022-03-22 18:44:43","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":313440,"visible":true,"origin":"","legend":"","description":"","filename":"R38supplementmateria.docx","url":"https://assets-eu.researchsquare.com/files/rs-1105606/v1/5f932f65261315365d608095.docx"},{"id":19496271,"identity":"5dfe5ff7-addf-4ad5-b41f-1d764d9b20ee","added_by":"auto","created_at":"2022-03-22 18:41:43","extension":"xlsx","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":12144,"visible":true,"origin":"","legend":"","description":"","filename":"SupplemetaryTableS3.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-1105606/v1/be56f8c7f8b8899601b85301.xlsx"}],"financialInterests":"","formattedTitle":"Description of Prasinibacter corallicola gen. nov., sp. nov., a zeaxanthin-producing bacterium isolated from stony coral Porites lutea","fulltext":[{"header":"Introduction","content":"\u003cp\u003eGlobal coral reefs have degraded dramatically in recent decades (Hughes et al. \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2003\u003c/span\u003e; Bellwood et al. \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2004\u003c/span\u003e) and massive corals have replaced branched corals as the dominant assemblages (Perry et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Yu et al. \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Thermal stress is one of the main causes of scleractinian coral degradation (Gardner et al. \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2003\u003c/span\u003e; Hughes et al. \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Hughes et al. \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Recently, it was reported that the phycosphere bacteria \u003cem\u003eMuricauda\u003c/em\u003e sp. GF1 protects coral endosymbionts from thermal stress by producing zeaxanthin (Motone et al. \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Zeaxanthin is usually synthesized in flavobacteria from terpenoids by the combinations of phytoene synthase (CrtB), phytoene dehydrogenase (CrtI), lycopene cyclase (CrtY), and β-carotene hydroxylase (CrtZ) (Zhang et al. \u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). According to the List of Prokaryotic names with Standing in Nomenclature (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://lpsn.dsmz.de/\u003c/span\u003e\u003cspan address=\"https://lpsn.dsmz.de/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e), to date, up to 151 validly published genera have been assigned to the family \u003cem\u003eFlavobacteriaceae\u003c/em\u003e. Genomic and physiological analyses have indicated that flavobacteria can degrade a diverse range of carbohydrates and proteins (Bauer et al. \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; Qin et al. \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Gavriilidou et al. \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2020\u003c/span\u003e), and a few are animal pathogens (Duchaud et al. \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2007\u003c/span\u003e; Loch and Faisal \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Adamek et al. \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). To determine the roles of flavobacteria in coral health, pure cultures were isolated from thermal tolerant corals: \u003cem\u003ePorites lutea\u003c/em\u003e, \u003cem\u003eGalaxea fascicularis\u003c/em\u003e, and \u003cem\u003eFavia\u003c/em\u003e sp. The taxonomic position of a new coral-associated flavobacterium, strain R38\u003csup\u003eT\u003c/sup\u003e, was identified using polyphasic identification, and the results indicated that strain R38\u003csup\u003eT\u003c/sup\u003e and other coral-associated flavobacteria can also produce zeaxanthin. Therefore, coral-associated flavobacteria may help alleviate host thermal stress caused by global warming.\u003c/p\u003e"},{"header":"Materials And Methods","content":"\u003ch2\u003eIsolation, cultivation, and maintenance\u003c/h2\u003e\n\u003cp\u003eMassive coral \u003cem\u003eP. lutea\u003c/em\u003e was collected from Weizhou Island (109\u0026ordm; 08\u0026prime; 35\u0026prime;\u0026prime; E, 21\u0026ordm; 03\u0026prime; 42\u0026prime;\u0026prime; N), in the Beibu Gulf, China.\u0026nbsp;\u003cem\u003eG. fascicularis\u0026nbsp;\u003c/em\u003eand \u003cem\u003eFavia\u003c/em\u003e sp\u003cem\u003e.\u003c/em\u003e were collected from Daya Bay (114\u0026ordm; 38\u0026prime; 32 \u0026prime;\u0026prime;E, 22\u0026ordm; 34\u0026prime; 32\u0026prime;\u0026prime; N), China. The coral\u0026nbsp;pieces (approximately 1 cm\u0026nbsp;\u0026times;\u0026nbsp;1 cm) were washed twice using sterile natural seawater,\u0026nbsp;then the coral tissue was homogenized using silica beads on vortex mixer after\u0026nbsp;scraping with scissors. The homogenate was diluted by ten-fold using sterile seawater, and 100 \u0026mu;l of each dilution was spread on\u0026nbsp;modified R2A plate (marine R2A plate, R2A agar was obtained from BD, which was dissolved in natural seawater, pH7.6). Bacteria\u0026nbsp;were incubated at 25 \u0026deg;C for two weeks. Colonies were picked and purified on marine R2A plates, and stored at -70 \u0026deg;C in R2A broth (Haibo, China)/glycerol (4:1, v/v).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003ePreliminary identification using 16S rRNA gene sequence indicated\u0026nbsp;that\u0026nbsp;strain R38\u003csup\u003eT\u003c/sup\u003e (from \u003cem\u003eP. lutea\u003c/em\u003e)\u003cem\u003e\u0026nbsp;\u003c/em\u003emay\u003cem\u003e\u0026nbsp;\u003c/em\u003erepresent a new species in a new genus, therefore, polyphasic identification was performed to identify its exact taxonomic position. Type strains\u003cem\u003e\u0026nbsp;Leptobacterium flavescens\u0026nbsp;\u003c/em\u003eKCTC 22160\u003csup\u003eT\u003c/sup\u003e and \u003cem\u003eSpongiivirga citrea\u003c/em\u003e KCTC 32990\u003csup\u003eT\u003c/sup\u003e, obtained from the Korean Collection for Type Cultures (KCTC), were used as references. Both\u0026nbsp;the new isolate and the reference type strains grew well on marine agar 2216 (BD).\u003c/p\u003e\n\u003ch2\u003ePhysiological and biochemical analyses\u003c/h2\u003e\n\u003cp\u003eCellular morphology was observed using\u0026nbsp;an optical microscope (Olympus BX53) and transmission electron microscope (Tecnai G2 F30 S-TWIN/X-MAX 80) after incubation for 2-3 days in marine broth 2216 (BD) at 30 \u0026deg;C. Cell mobility was tested using the hanging drop technique (Bernardet et al. 2002). The Gram reaction was determined as described by Gerhardt et al. (1994). Catalase activity was determined by observing bubble production in a 3 % (v/v) hydrogen peroxide solution, and oxidase activity was determined using oxidase test strips (Huankai, China). The ability to form\u0026nbsp;endospores was examined as described by Dong \u0026amp; Cai (2001). NaCl requirement and tolerance were tested at 30 \u0026deg;C for 7 days in R2A liquid medium (Haibo, China) with NaCl concentrations ranging from 0-16% (w/v), namely 0, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, and 16 % (w/v). Growth at different pH, adjusted with different buffers (1\u0026nbsp;interval, pH 4-5, 0.1 M citrate; pH 6-8, 0.1 M phosphate; pH 9-11, 0.1 M carbonate),\u0026nbsp;was tested in R2A liquid medium [including 1% NaCl (w/v)] at 30 \u0026deg;C for 7 days.\u0026nbsp;The\u0026nbsp;optimal growth temperature for bacteria on marine agar 2216 (BD) plates was determined after 7-30 days of growth at temperatures\u0026nbsp;of 4, 10, 15, 20, 25, 30, 33, 37, and 40 \u0026deg;C. Metabolism ability was characterized\u0026nbsp;using API 20NE strips, API ZYM strips, and Biolog Gen III\u0026nbsp;microplates according to the manufacturer\u0026rsquo;s protocols. Anaerobic fermentation was determined using API 50CH strips according to the manufacturer\u0026rsquo;s protocol with the inoculum medium being marine broth 2216 (BD).\u003c/p\u003e\n\u003ch2\u003eChemotaxonomic characterization\u003c/h2\u003e\n\u003cp\u003eThe\u0026nbsp;biomass of strain R38\u003csup\u003eT\u003c/sup\u003e and\u0026nbsp;the reference strain for cellular fatty acid analysis\u0026nbsp;were acquired from the third quadrant of the streaked marine agar 2216 plate incubated at\u0026nbsp;28 \u0026deg;C. Cellular fatty acid composition was analyzed by gas chromatography (Agilent G6890N) and identified using the Sherlock Microbial Identification System (Version 6.0) according to the manufacturer\u0026rsquo;s instructions. Biomass for the analyses of quinones and polar lipids was obtained from marine broth 2216 after 3 days of incubation at 30 \u0026deg;C. Respiratory quinones were extracted as described by Collins\u0026nbsp;(1994) and analyzed using reversed-phase high-performance liquid chromatography (HPLC)\u0026nbsp;(Komagata \u0026amp; Suzuki, 1987).\u0026nbsp;The isoprenoid quinones were eluted using a mixture of methanol/2-propanol (2:1, v/v) and flow rate of 1 ml/min at room temperature and detected by UV absorbance at 270 nm.\u0026nbsp;Polar lipids were extracted as described by Kamekura (1993) and identified by spraying with ethanolic molybdophosphoric acid, molybdenum blue, ninhydrin, \u0026alpha;-naphthol/sulfuric acid, and Dragendorff\u0026apos;s reagent after two-dimensional thin layer chromatography (TLC) (Tindall\u0026nbsp;1990).\u0026nbsp;\u003c/p\u003e\n\u003ch2\u003ePhylogenetic and phylogenomic analysis\u003c/h2\u003e\n\u003cp\u003eThe 16S rRNA gene of strain R38\u003csup\u003eT\u003c/sup\u003e was obtained using PCR amplification with the universal primers 27F and 1492R (Lane 1991) and sequenced using the Sanger method. The 16S rRNA gene sequence similarities were determined using the EzBioCloud (Yoon et al.\u0026nbsp;2017) and NCBI database. Alignment of 16S rRNA gene sequences was performed using the SINA software package (Pruesse et al.\u0026nbsp;2012) and\u0026nbsp;the SILVA rRNA database. Phylogenetic trees were constructed using the maximum likelihood (Felsenstein 1981), neighbor-joining (Saitou \u0026amp; Nei 1987), and maximum-parsimony (Swofford\u0026nbsp;1993) algorithms in the software package MEGA version 7.0 (Kumar et al.\u0026nbsp;2016). The phylogenetic distance matrices were estimated using the Kimura two-parameter model (Kimura,\u0026nbsp;1980). The topology of the phylogenetic tree was analyzed using the bootstrap resampling method of Felsenstein (1985) with 1000 replicates. Whole-genome sequencing was performed using an Illumina HiSeq PE150 platform. Library construction was performed by\u0026nbsp;PCR amplification of a 350 bp insert\u0026nbsp;with A-tail ligated to paired-end adaptors,\u0026nbsp;at the Beijing Novogene Bioinformatics Technology Co., Ltd. Good-quality paired reads were assembled into a number of scaffolds using the SOAPdenovo (Li et al. 2008; Li et al.\u0026nbsp;2010). Genomic information was extracted as described by Chun et al. (Chun et al.\u0026nbsp;2018). The phylogenetic tree was reconstructed using the up-to-date bacterial core gene set (UBCG v.3) according to the manual (Na et al.\u0026nbsp;2018). The average nucleotide identity (ANI) was calculated using the online ANI calculator (Yoon et al. 2017). The average amino acid identity (AAI) was calculated using\u0026nbsp;the EzAAI (Kim et al.\u0026nbsp;2021). Carbohydrate-active enzyme annotation was performed using the dbCAN meta server (Zhang et al. 2018). Peptidases were annotated using the Hotpep-protease method (Bush, 2020) based on the Merops database.\u0026nbsp;\u003c/p\u003e\n\u003ch2\u003eZeaxanthin detection\u003c/h2\u003e\n\u003cp\u003eThe production of\u0026nbsp;zeaxanthin by bacteria from the family \u003cem\u003eFlavobacteriaceae\u003c/em\u003e was assessed using both genome annotation and HPLC assay. The existence of coding genes for zeaxanthin biosynthesis enzymes such as phytoene desaturase, lycopene beta-cyclase, and beta-carotene 3-hydroxylase was checked using the Joint Genome Institute online server (\u003ca href=\"https://genome.jgi.doe.gov/portal/\"\u003ehttps://genome.jgi.doe.gov/portal/\u003c/a\u003e) and the NCBI genome server (\u003ca href=\"https://www.ncbi.nlm.nih.gov/genome/\"\u003ehttps://www.ncbi.nlm.nih.gov/genome/\u003c/a\u003e). Zeaxanthin was extracted from approximately 10 mg wet weight bacteria grown on an R2A plate using methanol and glass beads on a vortex for 30 seconds, and then analyzed using HPLC (Thermal Ultimate 3000). Samples were separated using an Agilent ZORBAX Eclipse XDB-C18 (250 mm, 5 \u0026micro;m particle size) at a column temperature of 35 \u0026deg;C, and the mobile phase comprised 90% (vol/vol) methanol containing 0.1% (v/v) formic acid at a flow rate of 1 ml/min (Motone et al. 2020). Zeaxanthin was checked using a photodiode array detector. Standard zeaxanthin was purchased from the Resource Platform of Standard Material (China). \u003c/p\u003e"},{"header":"Results And Discussion","content":"\u003ch2\u003e\u003cstrong\u003eZeaxanthin production\u003c/strong\u003e\u003c/h2\u003e\n\u003cp\u003eCoral bleaching caused by thermal stress is becoming increasingly serious (Hughes et al.\u0026nbsp;2018). In the last decade, coral bleaching has occurred at significantly higher sea surface temperatures (SSTs) (\u0026sim;0.5 \u0026deg;C) than in the previous decade, suggesting that thermally susceptible genotypes may have declined and/or adapted (Sully et al.\u0026nbsp;2019). Zeaxanthin is a carotenoid antioxidant that has been shown to protect endosymbiotic\u003cem\u003e\u0026nbsp;Symbiodiniaceae\u003c/em\u003e algae, isolated from coral \u003cem\u003eGalaxea fascicularis,\u003c/em\u003e from thermal and light stress (Motone et al. 2020). Coding genes for Zeaxanthin biosynthesis enzymes\u0026nbsp;phytoene desaturase, lycopene beta-cyclase,\u0026nbsp;and beta-carotene 3-hydroxylase\u0026nbsp;were examined in 115 of the 177 total genera in \u003cem\u003eFlavobacteriaceae\u003c/em\u003e, and over 50% of these genera (62) had all three enzymes (Table S1). Of note, all 11 strains of coral-associated flavobacteria (from approximately ten genera), including strain R38\u003csup\u003eT\u003c/sup\u003e, were able to produce zeaxanthin according to HPLC analysis (Table\u0026nbsp;1). These results indicate that the \u003cem\u003eFlavobacteriaceae\u003c/em\u003e family contains important zeaxanthin producers, and corals may benefit from these symbiotic flavobacteria when confronting thermal stress (Motone et al.\u0026nbsp;2020). However, zeaxanthin in coral (\u003cem\u003ePorites lutea\u003c/em\u003e and \u003cem\u003eAcropora\u003c/em\u003e sp.) or \u003cem\u003eSymbiodinium\u003c/em\u003e was below the limit of HPLC detection (Venn et al.\u0026nbsp;2006\u0026nbsp;and this study). Zeaxanthin is an intermediate product of\u0026nbsp;the algal accessory\u0026nbsp;photosynthetic pigments fucoxanthin and peridinin (Dautermann et al.\u0026nbsp;2020), and may be transformed immediately following synthesis in\u0026nbsp;the coral holobiont. This is suggested by the detection of\u0026nbsp;its precursor,\u0026nbsp;\u0026beta;-carotene, and the downstream product, peridinin, in coral or \u003cem\u003eSymbiodinium\u003c/em\u003e (Venn et al.\u0026nbsp;2006). Although zeaxanthin exchange between bacteria and\u0026nbsp;the endosymbiont\u0026nbsp;\u003cem\u003eSymbiodinium\u0026nbsp;\u003c/em\u003ehas not been shown, similar effects of pure culture \u003cem\u003eMuricauda\u003c/em\u003e sp. GF1 and zeaxanthin supplementation to cultured \u003cem\u003eSymbiodiniaceae\u003c/em\u003e conducted by Motone et al. (2020) strongly supports this exchange. Therefore, flavobacterial zeaxanthin may support\u0026nbsp;the\u0026nbsp;biosynthesis of algal accessory photosynthetic pigments in\u0026nbsp;the coral endosymbiont\u0026nbsp;\u003cem\u003eSymbiodinium\u003c/em\u003e. Finally, carotenoids can function as antioxidants via epoxidase/de-epoxidase reaction, regardless of the type of carotenoid\u0026nbsp;in\u0026nbsp;the final product (Krinsky\u0026nbsp;1989;\u0026nbsp;Lacour et al.\u0026nbsp;2020).\u003c/p\u003e\n\u003ch2\u003e\u003cstrong\u003eMorphological, physiological, and biochemical analyses\u003c/strong\u003e\u003c/h2\u003e\n\u003cp\u003eCells of bacterial strain R38\u003csup\u003eT\u003c/sup\u003e were gram-negative, non-spore-forming, non-motile, aerobic rods. Cells were usually 0.3-0.5 \u0026mu;m wide and 0.9-2.0 \u0026mu;m long (Fig.1), being narrower than that of \u003cem\u003eL. flavescens\u0026nbsp;\u003c/em\u003eKCTC 22160\u003csup\u003eT\u003c/sup\u003e, \u003cem\u003eS. citrea\u003c/em\u003e KCTC 32990\u003csup\u003eT\u003c/sup\u003e and \u003cem\u003eFulvibacter tottoriensis\u003c/em\u003e MTT-39\u003csup\u003eT\u003c/sup\u003e, while being wider than that of \u003cem\u003eMesoflavibacter aestuarii\u003c/em\u003e KYW614\u003csup\u003eT\u0026nbsp;\u003c/sup\u003e(Table 2). Cells of strain R38\u003csup\u003eT\u003c/sup\u003e could reduce nitrate to nitrogen, \u003cem\u003eL. flavescens\u0026nbsp;\u003c/em\u003eKCTC 22160\u003csup\u003eT\u003c/sup\u003eand \u003cem\u003eS. citrea\u003c/em\u003e KCTC 32990\u003csup\u003eT\u003c/sup\u003e could only reduce nitrate to nitrite, whereas\u003cem\u003e\u0026nbsp;M. aestuarii\u003c/em\u003e KYW614\u003csup\u003eT\u0026nbsp;\u003c/sup\u003e(Lee et al.\u0026nbsp;2014) and \u003cem\u003eF. tottoriensis\u003c/em\u003e MTT-39\u003csup\u003eT\u0026nbsp;\u003c/sup\u003e(Khan et al.\u0026nbsp;2008) could not reduce nitrate. Enzyme characterization of strain R38\u003csup\u003eT\u003c/sup\u003e using API ZYM strips showed a spectrum similar to that of \u003cem\u003eM. aestuarii\u003c/em\u003e KYW614\u003csup\u003eT\u003c/sup\u003e and \u003cem\u003eF. tottoriensis\u003c/em\u003e MTT-39\u003csup\u003eT\u003c/sup\u003e with the absence of\u0026nbsp;\u0026beta;-galactosidase, \u0026beta;-glucuronidase, \u0026alpha;-glucosidase, \u0026beta;-glucosidase, N-acetyl-\u0026beta;-glucosaminidase, and \u0026alpha;-mannosidase (Table 2) (Lee et al. 2014; Yoon et al.\u0026nbsp;2013). These enzyme results differed from those of\u0026nbsp;\u003cem\u003eL. flavescens\u0026nbsp;\u003c/em\u003eKCTC 22160\u003csup\u003eT\u0026nbsp;\u003c/sup\u003eand \u003cem\u003eS. citrea\u003c/em\u003e KCTC 32990\u003csup\u003eT\u0026nbsp;\u003c/sup\u003e(Table\u0026nbsp;2). Other characteristics of strain R38\u003csup\u003eT\u003c/sup\u003e are listed in Table\u0026nbsp;2 and the species description.\u003c/p\u003e\n\u003ch2\u003e\u003cstrong\u003eChemotaxonomic characteristics\u003c/strong\u003e\u003c/h2\u003e\n\u003cp\u003eThe only respiratory quinone detected in strain R38\u003csup\u003eT\u003c/sup\u003e was menaquinone 6 (MK-6), similar to \u003cem\u003eL. flavescens\u003c/em\u003e KCTC 22160\u003csup\u003eT\u0026nbsp;\u003c/sup\u003e(Mitra et al.\u0026nbsp;2009), \u003cem\u003eS. citrea\u003c/em\u003e KCTC 32990\u003csup\u003eT\u0026nbsp;\u003c/sup\u003e(Yoon et al.\u0026nbsp;2015), \u003cem\u003eF. tottoriensis\u003c/em\u003e MTT-39\u003csup\u003eT\u0026nbsp;\u003c/sup\u003e(Khan et al. 2008), and \u003cem\u003eMesoflavibacter aestuarii\u003c/em\u003e KYW614\u003csup\u003eT\u0026nbsp;\u003c/sup\u003e(Lee et al. 2014). Strain R38\u003csup\u003eT\u003c/sup\u003e contained\u0026nbsp;iso-C15 : 0\u0026nbsp;(42.4 %),\u0026nbsp;iso-C15 : 0\u0026omega;6c\u0026nbsp;(15.6 %),\u0026nbsp;C\u003csub\u003e16:2\u003c/sub\u003e DMA (7.8 %), and C\u003csub\u003e13:1\u003c/sub\u003e\u0026omega;3c (5.2 %) as the major cellular fatty acids, this profile was highly similar to that of\u0026nbsp;\u003cem\u003eL. flavescens\u0026nbsp;\u003c/em\u003eKCTC 22160\u003csup\u003eT\u003c/sup\u003e, although the proportion of individual components varied (Table S2). However, the individual fatty acid content of \u003cem\u003eS. citrea\u003c/em\u003e KCTC 32990\u003csup\u003eT\u003c/sup\u003e differed from that of strains R38\u003csup\u003eT\u003c/sup\u003e and \u003cem\u003eL. flavescens\u0026nbsp;\u003c/em\u003eKCTC 22160\u003csup\u003eT\u0026nbsp;\u003c/sup\u003e(Table\u0026nbsp;S2). The major fatty acids of strain R38\u003csup\u003eT\u003c/sup\u003e, \u003cem\u003eL. flavescens\u0026nbsp;\u003c/em\u003eKCTC 22160\u003csup\u003eT\u003c/sup\u003e,\u0026nbsp;and \u003cem\u003eS. citrea\u003c/em\u003e KCTC 32990\u003csup\u003eT\u003c/sup\u003e were highly different from those of \u003cem\u003eM. aestuarii\u003c/em\u003e KYW614\u003csup\u003eT\u003c/sup\u003e and \u003cem\u003eF. tottoriensis\u003c/em\u003e MTT-39\u003csup\u003eT\u0026nbsp;\u003c/sup\u003e(Table\u0026nbsp;2) (Lee et al.\u0026nbsp;2014; Yoon et al. 2013). The major polar lipids of strain R38\u003csup\u003eT\u003c/sup\u003e were phosphatidyl-N-methylethanolamine, phosphatidylethanolamine, one unidentified ninhydrin phospholipid, three unidentified ninhydrin-positive lipids, and three unidentified lipids (Supplementary Fig.S1). This polar lipid profile was similar to that of \u003cem\u003eL. flavescens\u0026nbsp;\u003c/em\u003eKCTC 22160\u003csup\u003eT\u003c/sup\u003e, but highly different from that of \u003cem\u003eS. citrea\u003c/em\u003e KCTC 32990\u003csup\u003eT\u003c/sup\u003e, which contains few types of polar lipids (Supplementary Fig.S1). However, strain R38\u003csup\u003eT\u003c/sup\u003e could still be distinguished from \u003cem\u003eL. flavescens\u0026nbsp;\u003c/em\u003eKCTC 22160\u003csup\u003eT\u003c/sup\u003e by its unidentified ninhydrin phospholipid (Supplementary Fig.S1). Furthermore, no phosphatidyl-N-methylethanolamine\u0026nbsp;has been reported for\u0026nbsp;\u003cem\u003eM. aestuarii\u003c/em\u003e KYW614\u003csup\u003eT\u003c/sup\u003e and \u003cem\u003eF. tottoriensis\u003c/em\u003e MTT-39\u003csup\u003eT\u0026nbsp;\u003c/sup\u003e(Lee et al. 2014; Yoon et al.\u0026nbsp;2013). Therefore,\u0026nbsp;the polar lipid profile distinguishes strain R38\u003csup\u003eT\u003c/sup\u003e from any validly published taxon.\u0026nbsp;\u003c/p\u003e\n\u003ch2\u003e\u003cstrong\u003eMolecular characterization and phylogenetic analysis\u003c/strong\u003e\u003c/h2\u003e\n\u003cp\u003eA\u0026nbsp;nearly complete 16S rRNA gene sequence (1383 nt) of strain R38\u003csup\u003eT\u003c/sup\u003e was obtained by Sanger sequencing and deposited in GenBank under accession number MN908337. Global alignment using\u0026nbsp;the EzBioCloud database indicated that the\u0026nbsp;most closely related neighbor of strain R38\u003csup\u003eT\u003c/sup\u003e is \u003cem\u003eMesoflavibacter aestuarii\u003c/em\u003e KYW614\u003csup\u003eT\u003c/sup\u003e, with a 16S rRNA gene similarity of 93%. The next most similar members were of \u003cem\u003eBizionia\u003c/em\u003e, \u003cem\u003eSabulilitoribacter\u003c/em\u003e, \u003cem\u003eGaetbulibacter\u003c/em\u003e,\u0026nbsp;and \u003cem\u003eAlgibacter\u003c/em\u003e genera which showed 92.7-92.9% sequence similarity. However, 16S rRNA gene phylogenetic analysis based on\u0026nbsp;the maximum-likelihood algorithm indicated that strain R38\u003csup\u003eT\u003c/sup\u003e forms a distinct branch in a stable cluster composed of strain R38\u003csup\u003eT\u003c/sup\u003e and \u003cem\u003eL. flavescens\u0026nbsp;\u003c/em\u003eKCTC 22160\u003csup\u003eT\u0026nbsp;\u003c/sup\u003e(Fig. 2). Neighbor-joining clustering also supports this tree topology (Supplementary Fig. S2). The close relationship between strain R38\u003csup\u003eT\u003c/sup\u003e and \u003cem\u003eL. flavescens\u0026nbsp;\u003c/em\u003eKCTC 22160\u003csup\u003eT\u003c/sup\u003e was also represented in the maximum-parsimony analysis, although a robust cluster was not formed (Supplementary Fig.S3).\u0026nbsp;\u003c/p\u003e\n\u003ch2\u003e\u003cstrong\u003eGenome properties and comparison\u003c/strong\u003e\u003c/h2\u003e\n\u003cp\u003eThe genome sequencing depth of strain R38\u003csup\u003eT\u003c/sup\u003e was 333\u0026times;,\u0026nbsp;and the N50 was 750154 bp. A total of 11 contigs were obtained, the obtained genome size was 3.94 Mb, and the\u0026nbsp;genomic DNA G+C content was 33.2 mol%. The genome sequencing depth of \u003cem\u003eL. flavescens\u003c/em\u003e KCTC 22160\u003csup\u003eT\u003c/sup\u003e was 154\u0026times;, the N50 was 1032064 bp, a total of 9 contigs were obtained, the obtained genome size was 4.21 Mb, and the genomic DNA G+C content was 40.9 mol%. The genome sequencing depth of \u003cem\u003eS. citrea\u003c/em\u003e KCTC 32990\u003csup\u003eT\u003c/sup\u003e was 155\u0026times;, the N50 was 637254 bp, a total of 17 contigs were obtained, the obtained genome size was 4.15 Mb, and the genomic DNA G+C content was 36.3 mol%.\u0026nbsp;The\u0026nbsp;genomes of closely related type strains were 3.05-5.03 MB, with G+C content of 33.2-55.3 mol% (Supplementary Table S3). Thus, strain R38\u003csup\u003eT\u003c/sup\u003e has the lowest genomic G+C content (33.2 mol%). The complete 16S rRNA gene of strain R38\u003csup\u003eT\u003c/sup\u003e obtained by genome sequencing was 1508 nt, and showed two nucleotide differences\u0026nbsp;compared to the sequence obtained by Sanger sequencing. Meanwhile, the result obtained by Sanger sequencing indicated there were at least two copies of the 16S rRNA gene in cells of strain R38\u003csup\u003eT\u003c/sup\u003e. The ANI of strain R38\u003csup\u003eT\u003c/sup\u003e to \u003cem\u003eL. flavescens\u003c/em\u003e KCTC 22160\u003csup\u003eT\u003c/sup\u003e,\u003cem\u003e\u0026nbsp;S. citrea\u003c/em\u003e KCTC 32990\u003csup\u003eT\u003c/sup\u003e,\u0026nbsp;and \u003cem\u003eM. aestuarii\u003c/em\u003e KYW614\u003csup\u003eT\u003c/sup\u003e were 72.5%, 69.6%, and 70.2%, respectively.\u0026nbsp;The AAI of strain R38\u003csup\u003eT\u003c/sup\u003e to \u003cem\u003eL. flavescens\u003c/em\u003e KCTC 22160\u003csup\u003eT\u0026nbsp;\u003c/sup\u003ewas 74.6%, and for other type strains, the indices were lower than 70%. Similar to the 16S rRNA gene-based phylogenetic results, phylogenomic analysis based on 92 genes also indicated\u0026nbsp;that strain R38\u003csup\u003eT\u003c/sup\u003e forms a distinct branch in a stable cluster composed of strain R38\u003csup\u003eT\u003c/sup\u003e and \u003cem\u003eL. flavescens\u003c/em\u003e KCTC 22160\u003csup\u003eT\u0026nbsp;\u003c/sup\u003e(Supplementary Fig.S4).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eApproximately 37 families of carbohydrate-active enzymes were detected in strain R38\u003csup\u003eT\u003c/sup\u003e, while in closely related type strains this number was between 39-100 (Supplementary Table S3). A limited quantity of glycoside hydrolases (16 vs 20-63) indicates\u0026nbsp;that strain R38\u003csup\u003eT\u0026nbsp;\u003c/sup\u003eis weak in carbohydrate utilization (Supplementary Table S3). Approximately 71 families of peptidases were detected in strain R38\u003csup\u003eT\u003c/sup\u003e. This quantity is higher than in most of the closely related type strains (Supplementary Table S3), indicating\u0026nbsp;that strain R38\u003csup\u003eT\u003c/sup\u003e is versatile in protein utilization. This carbohydrate and protein utilization pattern might have resulted from long-term bacteria-animal association.\u003c/p\u003e\n\u003ch2\u003e\u003cstrong\u003eTaxonomic conclusion\u003c/strong\u003e\u003c/h2\u003e\n\u003cp\u003eBased on phylogenetic analyses, strain R38\u003csup\u003eT\u0026nbsp;\u003c/sup\u003ewas found to be associated with the family \u003cem\u003eFlavobacteriaceae\u003c/em\u003e. The ANI of strain R38\u003csup\u003eT\u003c/sup\u003e to closely related type strains (\u0026le;72.5%) indicates\u0026nbsp;that strain R38\u003csup\u003eT\u003c/sup\u003e belongs to a novel species (Chun et al.\u0026nbsp;2018),\u0026nbsp;and both biochemical and chemotaxonomic characteristics (Table 2) support this species-level assignment. Furthermore, the low 16S rRNA gene similarities (\u0026le;93%) of strain R38\u003csup\u003eT\u003c/sup\u003e to closely related type strains indicate\u0026nbsp;that strain R38\u003csup\u003eT\u003c/sup\u003e represents a new genus (Yarza et al.\u0026nbsp;2014), which is also supported by the differences in polar lipid profile (Table\u0026nbsp;2\u0026nbsp;\u0026amp; Supplementary Fig.S1). Therefore, strain R38\u003csup\u003eT\u003c/sup\u003e represents a new species in a new genus under the family \u003cem\u003eFlavobacteriaceae\u003c/em\u003e, for which\u0026nbsp;\u003cem\u003ePrasinibacter corallicola\u0026nbsp;\u003c/em\u003egen. nov., sp. nov. is proposed.\u003c/p\u003e\n\u003ch2\u003e\u003cstrong\u003eDescription of\u003cem\u003e\u0026nbsp;Prasinibacter\u0026nbsp;\u003c/em\u003egen. nov.\u0026nbsp;\u003c/strong\u003e\u003c/h2\u003e\n\u003cp\u003e\u003cem\u003ePrasinibacter\u003c/em\u003e (Pra.si.ni.bac\u0026prime;ter. L. masc. adj. \u003cem\u003eprasinus\u003c/em\u003e, yellowish-green; N.L. masc. \u003cem\u003ebacter\u003c/em\u003e, rod: N.L. masc. n. \u003cem\u003eParisinibacter\u003c/em\u003e, a translucent yellowish green rod.)\u003c/p\u003e\n\u003cp\u003eCells are gram-negative, non-spore-forming, non-motile, aerobic rods. Catalase- and\u0026nbsp;oxidase-positive. Nitrate is reduced to nitrogen. The only menaquinone is MK-6. The major polar lipids are phosphatidyl-N-methylethanolamine, phosphatidylethanolamine, one unidentified ninhydrin phospholipid, three unidentified ninhydrin-positive lipids, and three unidentified lipids.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe type species is \u003cem\u003ePrasinibacter corallicola\u003c/em\u003e. Member of the family \u003cem\u003eFlavobacteriaceae\u003c/em\u003e.\u003c/p\u003e\n\u003ch2\u003e\u003cstrong\u003eDescription of \u003cem\u003ePrasibacter corallicola\u0026nbsp;\u003c/em\u003esp. nov.\u003c/strong\u003e\u003c/h2\u003e\n\u003cp\u003e\u003cem\u003ePrasinibacter corallicola\u003c/em\u003e (co.ral.li.co\u0026prime;la.\u0026nbsp;L. neut. n. \u003cem\u003ecorallum\u003c/em\u003e, coral; L. masc. suff. -\u003cem\u003ecola\u003c/em\u003e, inhabitant dweller; N.L. n. \u003cem\u003ecorallicola\u003c/em\u003e, coral-dweller)\u003c/p\u003e\n\u003cp\u003eThe description is as for the genus with the following additional properties: Cells are usually 0.3-0.5 \u0026mu;m wide and 0.9-2.0 \u0026mu;m long. Colonies are yellow-green, circular, and smooth on marine agar 2216. Cells can grow at 15-33 \u0026deg;C (optimum 25-30\u0026deg;C), pH 5-10 (optimum 7-8) in 3-6% (w/v) NaCl (optimum 3-4%) in R2A liquid medium. Zeaxanthin is produced. Production of H\u003csub\u003e2\u003c/sub\u003eS does not occur. Starch is hydrolyzed. In the API 20NE test, nitrate reduction and protease are positive. In the API ZYM test, alkaline phosphatase, esterase (C4), esterase lipase (C8) and (C14), leucine arylamidase, valine arylamidase, cysteine arylamidase, trypsin, \u0026alpha;-chymotrypsin, acid phosphatase, naphthol-AS-BI phosphohydrolase,\u0026nbsp;and \u0026alpha;-galactosidase\u0026nbsp;are positive. In the Biolog Gen III microplate, dextrin, D-maltose,\u0026nbsp;\u0026alpha;-D-glucose, D-mannose, L-alanine, L-glutamic acid, L-histidine,\u0026nbsp;\u0026alpha;-keto-glutaric acid, L-malic acid, bromo-succinic acid, Tween 40, acetoacetic acid, and acetic acid are oxidized.\u0026nbsp;The major fatty acids are\u0026nbsp;iso-C15 : 0,\u0026nbsp;iso-C15 : 0\u0026omega;6c,\u0026nbsp;C\u003csub\u003e16:2\u003c/sub\u003e DMA, and C\u003csub\u003e13:1\u003c/sub\u003e\u0026omega;3c. The genomic DNA G+C ratio is 33.2 mol%.\u003c/p\u003e\n\u003cp\u003eThe type strain, R38\u003csup\u003eT\u003c/sup\u003e (=MCCC 1K03889\u003csup\u003eT\u003c/sup\u003e=KCTC 72444\u003csup\u003eT\u003c/sup\u003e) was isolated from stony coral \u003cem\u003ePorites lutea\u003c/em\u003e collected from Weizhou Island in the Beibu Gulf, China. The GenBank accession number of the 16S rRNA gene sequence of the type strain is MN908337.\u003c/p\u003e\n\u003ch2\u003e\u0026nbsp;\u003c/h2\u003e"},{"header":"Abbreviations","content":"\u003ctable border=\"1\" cellpadding=\"0\" cellspacing=\"0\" width=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"15.370705244122966%\"\u003e\n \u003cp\u003eKCTC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"84.62929475587704%\"\u003e\n \u003cp\u003eThe Korean Collection for Type Cultures\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"15.370705244122966%\"\u003e\n \u003cp\u003eMCCC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"84.62929475587704%\"\u003e\n \u003cp\u003eThe Marine Culture Collection of China\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"15.370705244122966%\"\u003e\n \u003cp\u003eANI\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"84.62929475587704%\"\u003e\n \u003cp\u003eAverage nucleotide identity\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"15.370705244122966%\"\u003e\n \u003cp\u003eQ\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"84.62929475587704%\"\u003e\n \u003cp\u003eUbiquinone\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"15.370705244122966%\"\u003e\n \u003cp\u003eDPG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"84.62929475587704%\"\u003e\n \u003cp\u003eDiphosphatidylglycerol\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"15.370705244122966%\"\u003e\n \u003cp\u003ePG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"84.62929475587704%\"\u003e\n \u003cp\u003ePhosphatidylglycerol\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"15.370705244122966%\"\u003e\n \u003cp\u003ePME\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"84.62929475587704%\"\u003e\n \u003cp\u003ephosphatidylmethylethanolamine\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"15.370705244122966%\"\u003e\n \u003cp\u003ePE\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"84.62929475587704%\"\u003e\n \u003cp\u003ePhosphatidylethanolamine\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"15.370705244122966%\"\u003e\n \u003cp\u003eNL/AL\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"84.62929475587704%\"\u003e\n \u003cp\u003eUnidentified ninhydrin positive lipid\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"15.370705244122966%\"\u003e\n \u003cp\u003eNPL\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"84.62929475587704%\"\u003e\n \u003cp\u003eUnidentified ninhydrin positive phospholipid\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"15.370705244122966%\"\u003e\n \u003cp\u003ePL\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"84.62929475587704%\"\u003e\n \u003cp\u003eUnidentified phospholipid\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"15.370705244122966%\"\u003e\n \u003cp\u003eL\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"84.62929475587704%\"\u003e\n \u003cp\u003eUnidentified lipid\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAuthor contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eGuanghua Wang, Jianfeng Liu and Yuanjin Li isolated strains, performed experiments and wrote the manuscript. Biao Chen and Zhiheng Liao collected samples. \u0026nbsp;Hongfei Su and Jiayuan Liang gave advices about bacteria cultivation. Kefu Yu conceived and designed the experiments and approved the final manuscript.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research was supported by the\u0026nbsp;National Natural Sciences Foundation of China (41866004, 42090041 and 42030502);\u0026nbsp;National Key R \u0026amp; D Program of China (2018YFD0900803); Science and Technology Project of Guangxi (AD17129063, AA17204074 and AA18242026).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDeclarations\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflicts of interest\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe have no conflict of interest to declare.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthical approval\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n \u003cli\u003eAdamek M, Teitge F, Jung-Schroers V, Heling M, Gela D, Piackova V, Kocour M, Steinhagen D. 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Glob Chang Biol\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e21(3): 1153-1164.\u003c/li\u003e\n \u003cli\u003ePruesse E, Peplies J, Gl\u0026ouml;ckner FO (2012) \u0026nbsp;SINA: accurate high-throughput multiple sequence alignment of ribosomal RNA genes. Bioinformatics\u003cstrong\u003e\u003cem\u003e\u0026nbsp;\u003c/em\u003e\u003c/strong\u003e28:1823-1829.\u003c/li\u003e\n \u003cli\u003eQin QL, Zhang XY, Wang XM, Liu GM, Chen XL, Xie BB, Dang HY, Zhou BC, Yu J, Zhang YZ. (2010) The complete genome of \u003cem\u003eZunongwangia profunda\u003c/em\u003e SM-A87 reveals its adaptation to the deep-sea environment and ecological role in sedimentary organic nitrogen degradation. BMC Genomics 11:247.\u003c/li\u003e\n \u003cli\u003eRosado PM, Leite DCA, Duarte GAS, Chaloub RM, Jospin G, Nunes da Rocha U, P Saraiva J, Dini-Andreote F, Eisen JA, Bourne DG, Peixoto RS (2019) Marine probiotics: increasing coral resistance to bleaching through microbiome manipulation.\u003cem\u003e\u0026nbsp;\u003c/em\u003eISME J 13(4): 921-936.\u003c/li\u003e\n \u003cli\u003eSaitou N \u0026amp; Nei M (1987)\u0026nbsp;The neighbor-joining method: a new method for reconstructing phylogenetic tree. Mol Biol Evol\u003cem\u003e\u0026nbsp;\u003c/em\u003e4: 406-425.\u003c/li\u003e\n \u003cli\u003eSully S, Burkepile DE, Donovan MK, Hodgson G, van Woesik R (2019) A global analysis of coral bleaching over the past two decades. Nat Commun 10(1): 1264.\u003c/li\u003e\n \u003cli\u003eSwofford DL (1993) PAUP: Phylogenetic analysis using parsimony, version 3.1.1. Champaign, IL: Illinois Natural History Survey.\u003c/li\u003e\n \u003cli\u003eTindall BJ\u0026nbsp;(1990)\u0026nbsp;Lipid composition of \u003cem\u003eHalobacterium lacusprofundi\u003c/em\u003e. FEMS Microbiol Lett\u003cem\u003e\u0026nbsp;\u003c/em\u003e66: 199-202.\u003c/li\u003e\n \u003cli\u003eVenn AA, Wilson MA, Trapido-Rosenthal HG, Keely BJ, Douglas AE (2006) The impact of coral bleaching on the pigment profile of the symbiotic alga, Symbiodinium. Plant Cell Environ 29(12): 2133-42\u003c/li\u003e\n \u003cli\u003eYarza P, Yilmaz P, Pruesse E, Gl\u0026ouml;ckner FO, Ludwig W, Schleifer KH, Whitman WB, Euz\u0026eacute;by J, Amann R, Rossell\u0026oacute;-M\u0026oacute;ra R (2014) Uniting the classification of cultured and uncultured bacteria and archaea using 16S rRNA gene sequences. 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Curr Microbiol 70(1): 51-57.\u003c/li\u003e\n \u003cli\u003eYoon JH, Lee MH, Jung YT (2013) \u003cem\u003ePseudofulvibacter geojedonensis\u0026nbsp;\u003c/em\u003egen. nov., sp. nov., a polysaccharide-degrading member of the family \u003cem\u003eFlavobacteriaceae\u003c/em\u003e isolated from seawater, and emended description of the genus \u003cem\u003eFulvibacter\u003c/em\u003e.\u0026nbsp;Int J Syst Evol Microbiol\u0026nbsp;63: 1696-1701.\u003c/li\u003e\n \u003cli\u003eYoon SH, Ha SM, Kwon S, Lim J, Kim Y, Seo H, Chun J (2017) Introducing EzBioCloud: A taxonomically united database of 16S rRNA and whole genome assemblies.\u0026nbsp;Int J Syst Evol Microbiol\u003cem\u003e\u0026nbsp;\u003c/em\u003e67: 1613-1617.\u003c/li\u003e\n \u003cli\u003eYoon SH, Ha SM, Lim JM, Kwon SJ and Chun J (2017) A large-scale evaluation of algorithms to calculate average nucleotide identity. Antonie van Leeuwenhoek. 110: 1281-1286.\u0026nbsp;\u003c/li\u003e\n \u003cli\u003eYu W, Wang W, Yu K, Wang Y, Huang X, Huang R, Liao Z, Xu S, Chen X (2019) Rapid decline of a relatively high latitude coral assemblage at Weizhou Island, northern South China Sea. Biodiversity and Conservation 28: 3925-3949.\u003c/li\u003e\n \u003cli\u003eZhang H, Yohe T, Huang L, Entwistle S, Wu P, Yang Z, et al. (2018) dbCAN2: a meta server for automated carbohydrate-active enzyme annotation. Nucleic Acids Res\u003cem\u003e\u0026nbsp;\u003c/em\u003e46(W1): W95-W101.\u003c/li\u003e\n\u003c/ol\u003e"},{"header":"Tables","content":"\u003cp style=\"text-align: center;\"\u003e\u003cstrong\u003eTable 1\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp style=\"text-align: center;\"\u003eCoral associated bacterial ability of zeaxanthin production\u0026nbsp;\u003c/p\u003e\n\u003cp style=\"text-align: center;\"\u003eR \u0026amp; B were from\u003cem\u003e\u0026nbsp;Porites lutea\u003c/em\u003e; F was from \u003cem\u003eFavia\u003c/em\u003e sp.; and K was from \u003cem\u003eGalaxea fascicularis.\u003c/em\u003e *About 500 bp 16S rRNA gene sequence was used in the 27f end. +, positive; w, weak positive.\u003c/p\u003e\n\u003ctable border=\"1\" cellpadding=\"0\" cellspacing=\"0\" style=\"border-collapse: collapse; margin: 0px auto;\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd width=\"27.305605786618443%\"\u003e\n \u003cp\u003eStrains (accession No.)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"42.67631103074141%\"\u003e\n \u003cp\u003eTaxonomic neighbor\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.370705244122966%\"\u003e\n \u003cp\u003eIdentity (%)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.64737793851718%\"\u003e\n \u003cp\u003eZeaxanthin\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"27.305605786618443%\"\u003e\n \u003cp\u003eR38\u003csup\u003eT\u003c/sup\u003e (MN908337)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"42.67631103074141%\"\u003e\n \u003cp\u003e\u003cem\u003eMesoflavibacter aestuarii\u003c/em\u003e KYW614\u003csup\u003eT\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.370705244122966%\"\u003e\n \u003cp\u003e93\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.64737793851718%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"27.305605786618443%\"\u003e\n \u003cp\u003eR33\u003csup\u003eT\u0026nbsp;\u003c/sup\u003e(MN908336)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"42.67631103074141%\"\u003e\n \u003cp\u003e\u003cem\u003ePoritiphilus flavus\u003c/em\u003e R33\u003csup\u003eT\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.370705244122966%\"\u003e\n \u003cp\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.64737793851718%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"27.305605786618443%\"\u003e\n \u003cp\u003eBMA10(MZ779023)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"42.67631103074141%\"\u003e\n \u003cp\u003e\u003cem\u003eFulvivirga lutimaris\u003c/em\u003e TM-6\u003csup\u003eT\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.370705244122966%\"\u003e\n \u003cp\u003e92.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.64737793851718%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"27.305605786618443%\"\u003e\n \u003cp\u003eBMA12(MZ779024)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"42.67631103074141%\"\u003e\n \u003cp\u003e\u003cem\u003eFulvivirga lutimaris\u003c/em\u003e TM-6\u003csup\u003eT\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.370705244122966%\"\u003e\n \u003cp\u003e90.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.64737793851718%\"\u003e\n \u003cp\u003ew\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"27.305605786618443%\"\u003e\n \u003cp\u003eB25(MZ779016)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"42.67631103074141%\"\u003e\n \u003cp\u003e\u003cem\u003eUlvibacter litoralis\u003c/em\u003e KMM 3912\u003csup\u003eT\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.370705244122966%\"\u003e\n \u003cp\u003e94.9*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.64737793851718%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"27.305605786618443%\"\u003e\n \u003cp\u003eF16(MZ779017)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"42.67631103074141%\"\u003e\n \u003cp\u003e\u003cem\u003eTenacibaculum skagerrakense\u003c/em\u003e D30\u003csup\u003eT\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.370705244122966%\"\u003e\n \u003cp\u003e96.9*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.64737793851718%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"27.305605786618443%\"\u003e\n \u003cp\u003eF26(MZ779019)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"42.67631103074141%\"\u003e\n \u003cp\u003e\u003cem\u003eAquimarina atlantica\u003c/em\u003e 22II-S11-z7\u003csup\u003eT\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.370705244122966%\"\u003e\n \u003cp\u003e100*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.64737793851718%\"\u003e\n \u003cp\u003ew\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"27.305605786618443%\"\u003e\n \u003cp\u003eF45(MZ779020)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"42.67631103074141%\"\u003e\n \u003cp\u003e\u0026lsquo;Gramella jeungdoensis\u0026rsquo; HMD3159\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.370705244122966%\"\u003e\n \u003cp\u003e96.9*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.64737793851718%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"27.305605786618443%\"\u003e\n \u003cp\u003eF53(MZ779021)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"42.67631103074141%\"\u003e\n \u003cp\u003e\u0026lsquo;Gaetbulibacter jejuensis\u0026rsquo; CNURIC 14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.370705244122966%\"\u003e\n \u003cp\u003e100*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.64737793851718%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"27.305605786618443%\"\u003e\n \u003cp\u003eK6(MZ779018)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"42.67631103074141%\"\u003e\n \u003cp\u003e\u003cem\u003eMesoflavibacter sabulilitoris\u003c/em\u003e GJMS-9\u003csup\u003eT\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.370705244122966%\"\u003e\n \u003cp\u003e99.7*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.64737793851718%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"27.305605786618443%\"\u003e\n \u003cp\u003eK18(MZ779022)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"42.67631103074141%\"\u003e\n \u003cp\u003e\u003cem\u003eTenacibaculum mesophilum\u003c/em\u003e NBRC 16307\u003csup\u003eT\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.370705244122966%\"\u003e\n \u003cp\u003e95.1*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.64737793851718%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cstrong\u003eTable 2\u003c/strong\u003e\u003c/p\u003e\n\u003cp style=\"text-align: center;\"\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003ePhenotypic characteristics of strain R38\u003csup\u003eT\u003c/sup\u003e and related type strains\u003c/p\u003e\n\u003cp style=\"text-align: center;\"\u003eStrains: 1, R38\u003csup\u003eT\u003c/sup\u003e; 2,\u003cem\u003e\u0026nbsp;Leptobacterium flavescens\u0026nbsp;\u003c/em\u003eKCTC 22160\u003csup\u003eT\u003c/sup\u003e; 3, \u003cem\u003eSpongiivirga citrea\u003c/em\u003e KCTC 32990\u003csup\u003eT\u003c/sup\u003e; 4, \u003cem\u003eMesoflavibacter aestuarii\u003c/em\u003e KYW614\u003csup\u003eT\u0026nbsp;\u003c/sup\u003e(data from Lee et al. 2014); 5, \u003cem\u003eFulvibacter tottoriensis\u003c/em\u003e MTT-39\u003csup\u003eT\u0026nbsp;\u003c/sup\u003e(data from Khan et al.\u0026nbsp;2008; Yoon et al.\u0026nbsp;2013). +, positive; -, negative; ND, unknown.\u003c/p\u003e\n\u003cdiv align=\"center\"\u003e\n \u003ctable border=\"1\" cellpadding=\"0\" cellspacing=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd width=\"18.81188118811881%\"\u003e\n \u003cp\u003eCharacteristics\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.933521923620933%\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.548797736916548%\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.275813295615276%\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e4\u003csup\u003e\u0026dagger;\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.144271570014144%\"\u003e\n \u003cp\u003e5\u003csup\u003eɸ\u003c/sup\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"18.81188118811881%\"\u003e\n \u003cp\u003eHabitat\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.933521923620933%\"\u003e\n \u003cp\u003eCoral\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.548797736916548%\"\u003e\n \u003cp\u003eMarine sponge\u0026Dagger;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.275813295615276%\"\u003e\n \u003cp\u003eMarine sponge\u0026dagger;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003eSeawater\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.144271570014144%\"\u003e\n \u003cp\u003emarine sediment\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"18.81188118811881%\"\u003e\n \u003cp\u003eCell size (\u0026mu;m)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.933521923620933%\"\u003e\n \u003cp\u003e0.3-0.5\u0026times;0.9-2.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.548797736916548%\"\u003e\n \u003cp\u003e0.5-0.6\u0026times;8.5-9.0\u0026Dagger;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.275813295615276%\"\u003e\n \u003cp\u003e0.4-0.5\u0026times;2.0-3.0\u0026dagger;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e0.2\u0026times;1.2-1.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.144271570014144%\"\u003e\n \u003cp\u003e0.5-0.7\u0026times;5-14\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"18.81188118811881%\"\u003e\n \u003cp\u003eColony color\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.933521923620933%\"\u003e\n \u003cp\u003eYellow-green\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.548797736916548%\"\u003e\n \u003cp\u003ePale-yellow\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.275813295615276%\"\u003e\n \u003cp\u003eLemon-yellow\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003eOrange\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.144271570014144%\"\u003e\n \u003cp\u003eyellowish brown\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"18.81188118811881%\"\u003e\n \u003cp\u003eTemperature range (℃)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.933521923620933%\"\u003e\n \u003cp\u003e15-33\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.548797736916548%\"\u003e\n \u003cp\u003e15-35\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.275813295615276%\"\u003e\n \u003cp\u003e15-35\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e4-35\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.144271570014144%\"\u003e\n \u003cp\u003e10-37\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"18.81188118811881%\"\u003e\n \u003cp\u003eNaCl tolerance (%, w/v)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.933521923620933%\"\u003e\n \u003cp\u003e3-6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.548797736916548%\"\u003e\n \u003cp\u003e0.5-6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.275813295615276%\"\u003e\n \u003cp\u003e1-5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e1-9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.144271570014144%\"\u003e\n \u003cp\u003e1-5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"18.81188118811881%\"\u003e\n \u003cp\u003epH range\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.933521923620933%\"\u003e\n \u003cp\u003e5-10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.548797736916548%\"\u003e\n \u003cp\u003e6-10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.275813295615276%\"\u003e\n \u003cp\u003e8-9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e6-8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.144271570014144%\"\u003e\n \u003cp\u003e6-10\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"18.81188118811881%\"\u003e\n \u003cp\u003eNitrate reduction\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.933521923620933%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.548797736916548%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.275813295615276%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.144271570014144%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"6\" width=\"100%\"\u003e\n \u003cp\u003e\u003cstrong\u003eAPI ZYM test\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"18.81188118811881%\"\u003e\n \u003cp\u003e\u0026beta;-galactosidase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.933521923620933%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.548797736916548%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.275813295615276%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.144271570014144%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"18.81188118811881%\"\u003e\n \u003cp\u003e\u0026beta;-glucuronidase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.933521923620933%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.548797736916548%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.275813295615276%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.144271570014144%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"18.81188118811881%\"\u003e\n \u003cp\u003e\u0026alpha;-glucosidase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.933521923620933%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.548797736916548%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.275813295615276%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.144271570014144%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"18.81188118811881%\"\u003e\n \u003cp\u003e\u0026beta;-glucosidase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.933521923620933%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.548797736916548%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.275813295615276%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.144271570014144%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"18.81188118811881%\"\u003e\n \u003cp\u003eN-acetyl-b-glucosaminidase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.933521923620933%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.548797736916548%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.275813295615276%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.144271570014144%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"18.81188118811881%\"\u003e\n \u003cp\u003e\u0026alpha;-mannosidase\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.933521923620933%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.548797736916548%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.275813295615276%\"\u003e\n \u003cp\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.144271570014144%\"\u003e\n \u003cp\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"18.81188118811881%\"\u003e\n \u003cp\u003eGenome size (Mb)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.933521923620933%\"\u003e\n \u003cp\u003e3.94\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.548797736916548%\"\u003e\n \u003cp\u003e4.21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.275813295615276%\"\u003e\n \u003cp\u003e4.15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e2.92\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.144271570014144%\"\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"18.81188118811881%\"\u003e\n \u003cp\u003eANI of strain R38\u003csup\u003eT\u0026nbsp;\u003c/sup\u003eto\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.933521923620933%\"\u003e\n \u003cp\u003e100%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.548797736916548%\"\u003e\n \u003cp\u003e72.5%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.275813295615276%\"\u003e\n \u003cp\u003e69.6%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e70.2%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.144271570014144%\"\u003e\n \u003cp\u003eND\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"18.81188118811881%\"\u003e\n \u003cp\u003eG+C mol%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.933521923620933%\"\u003e\n \u003cp\u003e33.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.548797736916548%\"\u003e\n \u003cp\u003e40.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.275813295615276%\"\u003e\n \u003cp\u003e36.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e32.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.144271570014144%\"\u003e\n \u003cp\u003e35\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"18.81188118811881%\"\u003e\n \u003cp\u003eMajor polar lipid\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.933521923620933%\"\u003e\n \u003cp\u003ePME,PE,NLs,NPL,Ls\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.548797736916548%\"\u003e\n \u003cp\u003ePME,PE,NLs, Ls\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.275813295615276%\"\u003e\n \u003cp\u003ePME,NL,L\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003ePE, 2ALs, 3Ls\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.144271570014144%\"\u003e\n \u003cp\u003ePL,NLs,Ls\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd width=\"18.81188118811881%\"\u003e\n \u003cp\u003eMajor cellular fatty acids\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"20.933521923620933%\"\u003e\n \u003cp\u003eiso-C15 : 0, iso-C15 : 0\u0026omega;6c,\u0026nbsp;C\u003csub\u003e16:2\u003c/sub\u003e DMA, C\u003csub\u003e13:1\u003c/sub\u003e\u0026omega;3c\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"16.548797736916548%\"\u003e\n \u003cp\u003eiso-C15 : 0, iso-C15 : 0\u0026omega;6c,\u0026nbsp;C\u003csub\u003e16:2\u003c/sub\u003e DMA, C16 : 1\u0026omega;6c\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.275813295615276%\"\u003e\n \u003cp\u003eiso-C15 : 0, iso-C15 : 0\u0026omega;6c,\u0026nbsp;C\u003csub\u003e16:2\u003c/sub\u003e DMA, C\u003csub\u003e15:0\u003c/sub\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003eC15 : 0, iso-C15 : 0, iso-C16 : 0 3-OH, iso-C17 : 0 3-OH, iso-C15 : 1 G, C16 : 1 \u0026omega;7c/\u0026omega;6c\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.144271570014144%\"\u003e\n \u003cp\u003eiso-C\u003csub\u003e15 : 1\u003c/sub\u003e,C\u003csub\u003e16:0\u003c/sub\u003e3-OH,\u0026nbsp;iso-C\u003csub\u003e17 : 0\u003c/sub\u003e 3-OH, iso-C\u003csub\u003e15 :0\u0026nbsp;\u003c/sub\u003e3-OH, iso-C\u003csub\u003e15 : 0\u003c/sub\u003e.\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u0026Dagger;Data from Mitra et al. (2009); \u0026dagger; Data from Yoon et al. (2015).\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":true,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"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":"Prasinibacter corallicola, polyphasic taxonomy, 16S rRNA gene, zeaxanthin","lastPublishedDoi":"10.21203/rs.3.rs-1105606/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-1105606/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThermal stress is considered one of the main causes of mass scleractinian coral degradation; however, it is still unknown how corals can adapt to future global warming. In this study, ten genera of coral-associated flavobacteria were shown to produce zeaxanthin, a carotenoid antioxidant, which may help coral holobionts to alleviate thermal stress. In addition, a novel zeaxanthin-producing flavobacterium, designated R38\u003csup\u003eT\u003c/sup\u003e, was identified using polyphasic taxonomy. Although strain R38\u003csup\u003eT\u003c/sup\u003e shared a maximum 16S rRNA gene sequence similarity of 93% with \u003cem\u003eMesoflavibacter aestuarii\u003c/em\u003e KYW614\u003csup\u003eT\u003c/sup\u003e, phylogenetic analyses based on whole genome and 16S rRNA gene sequences revealed that strain R38\u003csup\u003eT\u003c/sup\u003e forms a distinct branch in a robust cluster composed of strain R38\u003csup\u003eT\u003c/sup\u003e and \u003cem\u003eLeptobacterium flavescens\u003c/em\u003e KCTC 22160\u003csup\u003eT\u003c/sup\u003e under the family \u003cem\u003eFlavobacteriaceae\u003c/em\u003e. Strain R38\u003csup\u003eT\u003c/sup\u003e exhibited average nucleotide identities of 70.2% and 72.5% for \u003cem\u003eM. aestuarii\u003c/em\u003e KYW614\u003csup\u003eT\u003c/sup\u003e and \u003cem\u003eL. flavescens\u003c/em\u003e KCTC 22160\u003csup\u003eT\u003c/sup\u003e, respectively. The only detected respiratory quinone was menaquinone 6 (MK-6). The genomic DNA G\u0026thinsp;+\u0026thinsp;C content was 33.2 mol%. The major polar lipids were phosphatidylmethylethanolamine, phosphatidylethanolamine, one unidentified ninhydrin phospholipid, three unidentified ninhydrin-positive lipids, and three unidentified lipids. The major cellular fatty acids were iso-C\u003csub\u003e15 : 0,\u003c/sub\u003e iso-C\u003csub\u003e15 : 0\u003c/sub\u003eω6c, C\u003csub\u003e16:2\u003c/sub\u003e DMA, and C\u003csub\u003e13:1\u003c/sub\u003eω3c. The distinct biochemical, chemotaxonomic, phylogenetic, and phylogenomic differences from validly published taxa suggest that strain R38\u003csup\u003eT\u003c/sup\u003e represents a new species of a new genus, for which \u003cem\u003ePrasinibacter corallicola\u003c/em\u003e gen. nov., sp. nov. is proposed. The type strain R38\u003csup\u003eT\u003c/sup\u003e (=\u0026thinsp;MCCC 1K03889\u003csup\u003eT\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;KCTC 72444\u003csup\u003eT\u003c/sup\u003e).\u003c/p\u003e","manuscriptTitle":"Description of Prasinibacter corallicola gen. nov., sp. nov., a zeaxanthin-producing bacterium isolated from stony coral Porites lutea","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2022-03-22 18:41:41","doi":"10.21203/rs.3.rs-1105606/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"editorInvitedReview","content":"","date":"2022-03-21T04:53:36+00:00","index":0,"fulltext":""},{"type":"reviewersInvited","content":"","date":"2022-03-20T03:20:36+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2022-03-03T10:35:06+00:00","index":"","fulltext":""},{"type":"submitted","content":"Antonie van Leeuwenhoek","date":"2022-03-02T07:25:25+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":"62023910-a8d6-45d0-8eff-93c66a87eda6","owner":[],"postedDate":"March 22nd, 2022","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2022-05-07T12:31:29+00:00","versionOfRecord":[],"versionCreatedAt":"2022-03-22 18:41:41","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-1105606","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-1105606","identity":"rs-1105606","version":["v1"]},"buildId":"-HB7Z8yhvgn0wM9Nzuekk","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}