Inoculation of wheat bran with nonpathogenic Fusarium species an effective biological control method for Fusarium crown and root rot in asparagus

Inoculation of wheat bran with nonpathogenic Fusarium species an effective biological control method for Fusarium crown and root rot in asparagus | 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 Inoculation of wheat bran with nonpathogenic Fusarium species an effective biological control method for Fusarium crown and root rot in asparagus Yosuke Maeda, Seishi Akino, Norio Kondo This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6986679/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 18 May, 2026 Read the published version in Microbes and Environments → Version 1 posted You are reading this latest preprint version Abstract Fusarium crown and root rot (FCRR), a disease caused by the fungus Fusarium oxysporum f. sp. asparagi, is one of the most destructive diseases affecting asparagus worldwide. Despite some evidence of the suppression of FCRR by several nonpathogenic F. oxysporum sensu lato strains in laboratory experiments, biological control trials in FCRR-infested fields have not consistently demonstrated the effectiveness of these strains. Therefore, we developed and assessed a novel method of using nonpathogenic F. oxsyporum strains for the biological control of FCRR in the field. Two strains (SM007 and FAoc5) effectively suppressed FCRR in young asparagus seedlings were cultured on wheat bran medium. Then, the material was then inoculated into mature second-year seedlings before transplanting them into infested soil. Both strains successfully suppressed FCRR. Next, we evaluated the impact of irrigation and on this system in greenhouse experiments utilizing SM007. We found that the density of the fungus colonizing in the wheat bran played a pivotal role in disease suppression. Both strains utilized in this study also exhibited the capacity to impede chlamydospore germination of the pathogen when glucose was incorporated into soil. This suggests that nutrient competition between nonpathogenic strains and the pathogen plays a crucial role in disease suppression, highlighting the importance of maintaining an optimal balance between these microorganisms in organic matter-based applications of biocontrol agents. Fusarium oxysporum Asparagus Biological control Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Full Text Supplementary Files SupplementaryMaterials.docx Cite Share Download PDF Status: Published Journal Publication published 18 May, 2026 Read the published version in Microbes and Environments → Version 1 posted 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. 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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-6986679","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":481791367,"identity":"071e418b-9279-427e-9f91-05cf4548abc0","order_by":0,"name":"Yosuke Maeda","email":"data:image/png;base64,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","orcid":"","institution":"Nogyo Shokuhin Sangyo Gijutsu Sogo Kenkyu Kiko","correspondingAuthor":true,"prefix":"","firstName":"Yosuke","middleName":"","lastName":"Maeda","suffix":""},{"id":481791368,"identity":"8034de15-572c-49f4-a1fb-f171efbb5bfe","order_by":1,"name":"Seishi Akino","email":"","orcid":"","institution":"Hokkaido University School of Agriculture Graduate School of Agriculture Research Faculty of Agriculture: Hokkaido Daigaku Nogakubu Daigakuin Nogaku Kenkyuin Nogakuin","correspondingAuthor":false,"prefix":"","firstName":"Seishi","middleName":"","lastName":"Akino","suffix":""},{"id":481791369,"identity":"66c6811a-99a7-4a38-ac7e-4a695f45d0e8","order_by":2,"name":"Norio Kondo","email":"","orcid":"","institution":"Hokkaido University: Hokkaido Daigaku","correspondingAuthor":false,"prefix":"","firstName":"Norio","middleName":"","lastName":"Kondo","suffix":""}],"badges":[],"createdAt":"2025-06-26 23:09:09","currentVersionCode":1,"declarations":{"humanSubjects":false,"vertebrateSubjects":false,"conflictsOfInterestStatement":false,"humanSubjectEthicalGuidelines":false,"humanSubjectConsent":false,"humanSubjectClinicalTrial":false,"humanSubjectCaseReport":false,"vertebrateSubjectEthicalGuidelines":false},"doi":"10.21203/rs.3.rs-6986679/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6986679/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1264/jsme2.ME25063","type":"published","date":"2026-05-19T00:00:00+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":86358177,"identity":"bbeb517a-866f-449b-96f2-fc57626ae646","added_by":"auto","created_at":"2025-07-09 17:46:30","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":466896,"visible":true,"origin":"","legend":"\u003cp\u003eDisease incidence of FCRR in asparagus seedlings after transplanting them into pathogen-infested soil. Each treatment involved the application of a nonpathogenic strain through bud-cell suspension irrigation and /or inoculated wheat bran incorporation. Each plot represents the mean value of each treatment at each dpi, except for NC. * and *** indicate the significant differences (\u003cem\u003eP \u003c/em\u003e\u0026lt; \u003cem\u003e0.05 \u003c/em\u003eand \u003cem\u003eP \u003c/em\u003e\u0026lt; \u003cem\u003e0.001\u003c/em\u003e, respectively) from PC, based on Dunnett’s test for AUDPC values.\u003c/p\u003e","description":"","filename":"Fig1.png","url":"https://assets-eu.researchsquare.com/files/rs-6986679/v1/925b4b71227b5d8ca0c65afb.png"},{"id":86358179,"identity":"3b61292c-faa5-4192-9582-82921349d835","added_by":"auto","created_at":"2025-07-09 17:46:30","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":749785,"visible":true,"origin":"","legend":"\u003cp\u003eDisease incidence of FCRR after the transplantation of asparagus seedlings into the pathogen-infested soil following the application of treated wheat bran (10 g/pot). Each plot represents the mean disease incidence for each treatment at each dpi. Results from two identical experiments are shown separately due to differences in the density of nonpathogenic strain SM007 colonized in wheat bran. Different characters indicate significant differences (\u003cem\u003eP \u003c/em\u003e\u0026lt; \u003cem\u003e0.05\u003c/em\u003e) based on Tukey-kramer’s test for AUDPC values (\u003cem\u003en \u003c/em\u003e= 4).\u003c/p\u003e","description":"","filename":"Fig2.png","url":"https://assets-eu.researchsquare.com/files/rs-6986679/v1/57446a485db5ee8e3b772b42.png"},{"id":86359123,"identity":"bfe48de6-5dda-4a79-a65c-d83768784f76","added_by":"auto","created_at":"2025-07-09 17:54:30","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":537717,"visible":true,"origin":"","legend":"\u003cp\u003eDisease incidence of FCRR following the transplantation of asparagus seedlings into pathogen-infested soil after inoculation with nonpathogenic strain SM007, either by irrigating bud-cell suspension or incorporating treated wheat bran. Each plot represents the mean disease incidence for each treatment at each dpi (\u003cem\u003en \u003c/em\u003e= 8).\u003c/p\u003e","description":"","filename":"Fig3.png","url":"https://assets-eu.researchsquare.com/files/rs-6986679/v1/4191cd3fa11804f4ec9f59ae.png"},{"id":86358184,"identity":"25bd5001-3ef2-4a1e-bcd0-9019943eed7b","added_by":"auto","created_at":"2025-07-09 17:46:30","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":398434,"visible":true,"origin":"","legend":"\u003cp\u003eChlamydospore germination rate of the pathogen (1.0 × 10\u003csup\u003e3\u003c/sup\u003e chlamydospores/g soil) when treated with 1 ml of bud-cell suspension of nonpathogenic strains (1.0 × 10\u003csup\u003e6\u003c/sup\u003e bud-cells/ml) at varying glucose levels. Each plot represents the mean germination rate, with error bars indicating standard errors. Significant interactions between strain treatment and glucose content (\u003cem\u003eP \u003c/em\u003e\u0026lt; \u003cem\u003e0.01\u003c/em\u003e) were detected based on logistic regression analysis.\u003c/p\u003e","description":"","filename":"Fig4.png","url":"https://assets-eu.researchsquare.com/files/rs-6986679/v1/b85bc2b43646eb0e1eaea2db.png"},{"id":86358186,"identity":"d519f955-ba54-494f-bd59-cb8c5eecda85","added_by":"auto","created_at":"2025-07-09 17:46:30","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":743057,"visible":true,"origin":"","legend":"\u003cp\u003eField experiments. (a) Above-ground disease incidence in each plot. (b) Ratio of plants exhibiting root lesion affecting more than 66% of the root system after cultivating mature plants in the Hokkaido University field following transplantation into pathogen-infested field soil and incorporation of wheat bran inoculated with each nonpathogenic strain. Different characters indicate significant differences (\u003cem\u003eP \u003c/em\u003e\u0026lt; \u003cem\u003e0.05\u003c/em\u003e) based on Tukey-kramer’s test for AUDPC values (\u003cem\u003en \u003c/em\u003e= 3). + and ** indicate significant effects (\u003cem\u003eP \u003c/em\u003e\u0026lt; \u003cem\u003e0.10 \u003c/em\u003eand \u003cem\u003eP \u003c/em\u003e\u0026lt; \u003cem\u003e0.01\u003c/em\u003e, respectively) on the number of severely diseased plants, as determined by logistic regression. NC and PC repesent negative and positive control, respectively.\u003c/p\u003e","description":"","filename":"Fig5.png","url":"https://assets-eu.researchsquare.com/files/rs-6986679/v1/dca656e6a691c6279ee8dd35.png"},{"id":109792686,"identity":"dae6d18a-8ac0-412b-a566-14a37ca74631","added_by":"auto","created_at":"2026-05-22 13:51:00","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":3294977,"visible":true,"origin":"","legend":"","description":"","filename":"Manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6986679/v1_covered_270028b6-d37a-4410-8086-581f4efe4554.pdf"},{"id":86358190,"identity":"8cd76ced-e782-40e9-9f74-a88de4e57970","added_by":"auto","created_at":"2025-07-09 17:46:30","extension":"docx","order_by":9,"title":"","display":"","copyAsset":false,"role":"supplement","size":98722,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementaryMaterials.docx","url":"https://assets-eu.researchsquare.com/files/rs-6986679/v1/e21b063c454cbc27638d25ea.docx"}],"financialInterests":"","formattedTitle":"\u003cp\u003eInoculation of wheat bran with nonpathogenic \u003cem\u003eFusarium \u003c/em\u003especies an effective biological control method for Fusarium crown and root rot in asparagus\u003c/p\u003e","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Fusarium oxysporum, Asparagus, Biological control","lastPublishedDoi":"10.21203/rs.3.rs-6986679/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6986679/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"Fusarium crown and root rot (FCRR), a disease caused by the fungus Fusarium oxysporum f. sp. asparagi, is one of the most destructive diseases affecting asparagus worldwide. Despite some evidence of the suppression of FCRR by several nonpathogenic F. oxysporum sensu lato strains in laboratory experiments, biological control trials in FCRR-infested fields have not consistently demonstrated the effectiveness of these strains. Therefore, we developed and assessed a novel method of using nonpathogenic F. oxsyporum strains for the biological control of FCRR in the field. Two strains (SM007 and FAoc5) effectively suppressed FCRR in young asparagus seedlings were cultured on wheat bran medium. Then, the material was then inoculated into mature second-year seedlings before transplanting them into infested soil. Both strains successfully suppressed FCRR. Next, we evaluated the impact of irrigation and on this system in greenhouse experiments utilizing SM007. We found that the density of the fungus colonizing in the wheat bran played a pivotal role in disease suppression. Both strains utilized in this study also exhibited the capacity to impede chlamydospore germination of the pathogen when glucose was incorporated into soil. This suggests that nutrient competition between nonpathogenic strains and the pathogen plays a crucial role in disease suppression, highlighting the importance of maintaining an optimal balance between these microorganisms in organic matter-based applications of biocontrol agents.","manuscriptTitle":"Inoculation of wheat bran with nonpathogenic Fusarium species an effective biological control method for Fusarium crown and root rot in asparagus","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-07-09 17:46:26","doi":"10.21203/rs.3.rs-6986679/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"4fcdea44-17e4-407e-925e-99d3c630c79d","owner":[],"postedDate":"July 9th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2026-05-22T13:50:53+00:00","versionOfRecord":{"articleIdentity":"rs-6986679","link":"https://doi.org/10.1264/jsme2.ME25063","journal":{"identity":"microbes-and-environments","isVorOnly":true,"title":"Microbes and Environments"},"publishedOn":"2026-05-19 00:00:00","publishedOnDateReadable":"May 19th, 2026"},"versionCreatedAt":"2025-07-09 17:46:26","video":"","vorDoi":"10.1264/jsme2.ME25063","vorDoiUrl":"https://doi.org/10.1264/jsme2.ME25063","workflowStages":[]},"version":"v1","identity":"rs-6986679","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6986679","identity":"rs-6986679","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}