Metabolite-mediated antagonism of endophytic bacteria against Curvularia lunata in banana revealed by multi-analytical profiling

Metabolite-mediated antagonism of endophytic bacteria against Curvularia lunata in banana revealed by multi-analytical profiling | 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 Metabolite-mediated antagonism of endophytic bacteria against Curvularia lunata in banana revealed by multi-analytical profiling Papan Chowhan, Shambhu Swarnakar, Sumi Paul, Arka Pratim Chakraborty This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8590626/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 14 You are reading this latest preprint version Abstract Curvularia lunata (MLP-01; GenBank accession ON246070) is an important phytopathogen that causes leaf spot disease in Musa paradisiaca , resulting in significant yield losses. In this study, two potent plant growth-promoting endophytic bacteria, Enterobacter cloacae (MRH-06; GenBank accession ON955844) and Achromobacter xylosoxidans (MRH-11; GenBank accession ON955872), isolated from healthy Musa sp. root tissues, were evaluated for their antifungal activity against C. lunata . Dual culture assays showed strong antagonism, with E. cloacae inhibiting 75% and A. xylosoxidans inhibiting 83% of mycelial growth. Extracellular metabolites were extracted with chloroform and ethyl acetate and analysed by gas chromatography–mass spectrometry (GC–MS) and Fourier transform infrared (FT-IR) spectroscopy. GC–MS profiling revealed a diverse range of bioactive metabolites, including phenolic compounds, fatty acids, long-chain hydrocarbons, esters, and nitrogen-containing heterocycles. FT-IR analysis confirmed the presence of hydroxyl, aliphatic, alkene, ester, ether, and amine functional groups, strongly supporting GC–MS-based compound identification. Nuclear Magnetic Resonance (NMR) was also used to identify the compounds. Along with transmission electron microscopy, confocal and fluorescence microscopic images were also taken to locate endophytic bacteria within the root tissue of Musa paradisiaca . Biolog GEN III phenotypic profile of strains MRH-06 and MRH-11 showed a high degree of similarity to Enterobacter cloacae and Achromobacterxylosoxidans ,respectively. HPLC quantification of trypthphan showed higher production of trypthophan by both the bacterial isolate which helps in Indole-3-Acetic Acid (IAA) production and growth promotion. The integrated analytical approach highlights the superior biocontrol potential of A. xylosoxidans compared with E. cloacae and demonstrates the role of endophytic bacterial metabolites as eco-friendly alternatives for the management of C. lunata in banana. Curvularia lunata Achromobacter xylosoxidans Enterobacter cloacae Endophytes GC–MS NMR FT-IR BIOLOG Confocal TEM Fluroscence HPLC Biocontrol Full Text Additional Declarations No competing interests reported. Cite Share Download PDF Status: Under Review Version 1 posted Editorial decision: Revision requested 27 Apr, 2026 Reviews received at journal 12 Apr, 2026 Reviews received at journal 10 Apr, 2026 Reviewers agreed at journal 06 Apr, 2026 Reviewers agreed at journal 02 Apr, 2026 Reviewers agreed at journal 19 Mar, 2026 Reviews received at journal 20 Feb, 2026 Reviewers agreed at journal 19 Feb, 2026 Reviewers agreed at journal 07 Feb, 2026 Reviewers invited by journal 05 Feb, 2026 Editor invited by journal 03 Feb, 2026 Editor assigned by journal 28 Jan, 2026 Submission checks completed at journal 28 Jan, 2026 First submitted to journal 28 Jan, 2026 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. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-8590626","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":587468089,"identity":"4e0d686b-beda-4f2c-9bad-ef8edbc393ca","order_by":0,"name":"Papan Chowhan","email":"","orcid":"","institution":"Raiganj University","correspondingAuthor":false,"prefix":"","firstName":"Papan","middleName":"","lastName":"Chowhan","suffix":""},{"id":587468090,"identity":"53367f60-29df-49e0-8034-03e20be1bac9","order_by":1,"name":"Shambhu Swarnakar","email":"","orcid":"","institution":"Raiganj University","correspondingAuthor":false,"prefix":"","firstName":"Shambhu","middleName":"","lastName":"Swarnakar","suffix":""},{"id":587468091,"identity":"ddf07c27-228c-48db-a9a6-98fbd0de28a1","order_by":2,"name":"Sumi Paul","email":"","orcid":"","institution":"Raiganj University","correspondingAuthor":false,"prefix":"","firstName":"Sumi","middleName":"","lastName":"Paul","suffix":""},{"id":587468092,"identity":"d26591d0-15b0-4513-8155-ed4e467c5389","order_by":3,"name":"Arka Pratim Chakraborty","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABAUlEQVRIiWNgGAWjYFACNgYGxgY4zwaIGRsPEKNFAspLA2lpIEnLYTCJVwv/7GOJHxh32NTxz+4x+/i17bzd2vbDQFtqbKJxaZE4l3ZYgvFMmoTEnTPGs2XbbidvO5MI1HIsLbcBl54z7A0SjG2HJRhu5BgzSwK1mB0AamFsOIxTi/wZ9uYfIC3yEC3nks3OP8SvxeAM2zGwLQZALYwf2w7Ymd0gYIvhGbY0i8QzaZIbb6QVMzOcS04wuwG0JQGPX+TOsBnf+LjDhl/uRvJmxh9ldvZm59MfPvhQY4Pb+yCQACY5DJh5GBgSGxAiBAH7A8YfDAz2xCkeBaNgFIyCkQQANtJjQ3R0smYAAAAASUVORK5CYII=","orcid":"","institution":"Raiganj University","correspondingAuthor":true,"prefix":"","firstName":"Arka","middleName":"Pratim","lastName":"Chakraborty","suffix":""}],"badges":[],"createdAt":"2026-01-13 10:24:09","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-8590626/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-8590626/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":102297445,"identity":"c5546d41-dcc1-4b4b-980e-b9b7008ca73a","added_by":"auto","created_at":"2026-02-10 10:27:27","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":11861902,"visible":true,"origin":"","legend":"","description":"","filename":"finalpaper.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8590626/v1_covered_df9f9499-8924-4ee2-8b40-fe1dd8fd6e4f.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Metabolite-mediated antagonism of endophytic bacteria against Curvularia lunata in banana revealed by multi-analytical profiling","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"discover-plants","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"Learn more about [Discover Plants](https://link.springer.com/journal/44372)","snPcode":"44372","submissionUrl":"https://submission.springernature.com/new-submission/44372/3","title":"Discover Plants","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Discover Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Curvularia lunata, Achromobacter xylosoxidans, Enterobacter cloacae, Endophytes, GC–MS, NMR, FT-IR, BIOLOG, Confocal, TEM, Fluroscence, HPLC, Biocontrol","lastPublishedDoi":"10.21203/rs.3.rs-8590626/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8590626/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e \u003cem\u003eCurvularia lunata\u003c/em\u003e (MLP-01; GenBank accession ON246070) is an important phytopathogen that causes leaf spot disease in \u003cem\u003eMusa paradisiaca\u003c/em\u003e, resulting in significant yield losses. 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FT-IR analysis confirmed the presence of hydroxyl, aliphatic, alkene, ester, ether, and amine functional groups, strongly supporting GC\u0026ndash;MS-based compound identification. Nuclear Magnetic Resonance (NMR) was also used to identify the compounds. Along with transmission electron microscopy, confocal and fluorescence microscopic images were also taken to locate endophytic bacteria within the root tissue of \u003cem\u003eMusa paradisiaca\u003c/em\u003e. Biolog GEN III phenotypic profile of strains MRH-06 and MRH-11 showed a high degree of similarity to \u003cem\u003eEnterobacter cloacae\u003c/em\u003e and \u003cem\u003eAchromobacterxylosoxidans\u003c/em\u003e,respectively. HPLC quantification of trypthphan showed higher production of trypthophan by both the bacterial isolate which helps in Indole-3-Acetic Acid (IAA) production and growth promotion. 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