Unravelling the secrets of wide hybridization among the brassica species

Unravelling the secrets of wide hybridization among the brassica species | 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 Unravelling the secrets of wide hybridization among the brassica species Shikha Joshi, Charu Bisht, Anu Singh, Anil Kumar, A. S. Jeena, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7715506/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Pre-fertilization barriers in Brassica and related species pose a significant challenge in interspecific and intergeneric hybridization, hindering the successful transfer of desirable genetic traits crucial for crop improvement. Pollen germination, pollen tube growth, and the tube's capacity to reach and fertilise the ovule are among multiple stages at which these obstacles may appear. This study focuses on identifying such pre-fertilization barriers and assessing meiotic abnormalities during microsporogenesis in interspecific hybrids of Brassica species. Pre-fertilization parameters such as pollen germination percentage, pollen tube growth, and pollen tube abnormalities were thoroughly examined. In interspecific hybrids, the mean pollen tube germination percentage was highest for B. carinata × B. nigra (33.67 ± 11.35) and lowest for B. carinata × B. napus (18.68 ± 6.62), whereas it was highest for B. napus × B. carinata (33.06 ± 13.41) in reciprocal crosses. B. carinata × B. nigra had the highest mean pollen tube growth (21.23 ± 5.33), followed by B. carinata × B. napus (10.18 ± 3.17), while B. nigra × B. carinata had the lowest mean growth among reciprocals (21.39 ± 7.53). Pollen tube abnormalities such as coiling, bending, hairpin shapes, tube tip swelling, bifurcation, multiple tube emergence, and misdirected growth were observed. The highest incidence of abnormalities occurred when B. carinata was used as a pollen parent with B. napus . Further observations using fluorescent microscopy revealed pre-fertilization incompatibilities in the stylar and ovule regions. Crossability data revealed varying percentages of seed set (ranging from 0% in B. nigra × B. carinata and B. rapa × B. carinata to 25.64% in B. carinata × B. nigra) and siliqua set (from 33% in B. carinata × B. napus to 72% in selfed B. carinata ). Additionally, meiotic analysis revealed abnormalities such as improper alignment during M-I, M-II, A-I, A-II, and Telophase, indicating cytological instability in some hybrids. Correlation study further established the association among all of these traits. This study highlights critical reproductive and cytological barriers affecting hybrid success in Brassica , offering valuable insights for breeders. By identifying suitable parent combinations and understanding hybridization limitations, it supports more efficient transfer of traits and the development of diverse, improved lines for future breeding programs. Interspecific hybridization Pre-fertilization barrier meiotic abnormalities Full Text Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted 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. 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-7715506","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":527069005,"identity":"fe8cd608-e6d6-4bda-866b-e74710b32696","order_by":0,"name":"Shikha Joshi","email":"","orcid":"","institution":"Govind Ballabh Pant University of Agriculture and Technology","correspondingAuthor":false,"prefix":"","firstName":"Shikha","middleName":"","lastName":"Joshi","suffix":""},{"id":527069006,"identity":"b339e055-bb1e-453d-a5c1-44e87089e32a","order_by":1,"name":"Charu Bisht","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA+klEQVRIiWNgGAWjYBACe2YGNiB1AIiZj//4AKTY2AloMWwGajkA1sKWIDkDRDET0GJwAK6Fx0CaByRESIthO/uzxx9q7sibz0hLMLb5tU2ej5mB8cPHHHx+4TE3OHDsmeGcG8kHknP7bhu2MTMwS87chs8vPGwSBxsOM86QSEs4nNtzmxGohY2ZF48Wg8Psz0Ba7GdI5Bg2W/bctidCC4MZSEsiUIsxM8OP24kEtQAdZiZx5tjh5Bk8z9IYextuJ7cxMzbj9Ys9//FnEhU1h21nsCcfY/jx57bt/Pbmgx8+4tGCChjbwGQDsepB4A8pikfBKBgFo2CkAACUsVPDY2IK6gAAAABJRU5ErkJggg==","orcid":"","institution":"Teerthanker Mahaveer University","correspondingAuthor":true,"prefix":"","firstName":"Charu","middleName":"","lastName":"Bisht","suffix":""},{"id":527069007,"identity":"fc29be96-dd18-432b-bd43-9318522c8706","order_by":2,"name":"Anu Singh","email":"","orcid":"","institution":"Govind Ballabh Pant University of Agriculture and Technology","correspondingAuthor":false,"prefix":"","firstName":"Anu","middleName":"","lastName":"Singh","suffix":""},{"id":527069008,"identity":"e8a6bfeb-2d98-474a-9c67-1c37915a6e1c","order_by":3,"name":"Anil Kumar","email":"","orcid":"","institution":"Govind Ballabh Pant University of Agriculture and Technology","correspondingAuthor":false,"prefix":"","firstName":"Anil","middleName":"","lastName":"Kumar","suffix":""},{"id":527069009,"identity":"e482f391-e46d-46c0-b2cd-1bdb03fcf8a7","order_by":4,"name":"A. 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species","fulltext":[],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"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":"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":"Interspecific hybridization, Pre-fertilization barrier, meiotic abnormalities","lastPublishedDoi":"10.21203/rs.3.rs-7715506/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7715506/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003ePre-fertilization barriers in \u003cem\u003eBrassica\u003c/em\u003e and related species pose a significant challenge in interspecific and intergeneric hybridization, hindering the successful transfer of desirable genetic traits crucial for crop improvement. Pollen germination, pollen tube growth, and the tube's capacity to reach and fertilise the ovule are among multiple stages at which these obstacles may appear. This study focuses on identifying such pre-fertilization barriers and assessing meiotic abnormalities during microsporogenesis in interspecific hybrids of \u003cem\u003eBrassica\u003c/em\u003e species. Pre-fertilization parameters such as pollen germination percentage, pollen tube growth, and pollen tube abnormalities were thoroughly examined. In interspecific hybrids, the mean pollen tube germination percentage was highest for \u003cem\u003eB. carinata\u003c/em\u003e \u0026times; \u003cem\u003eB. nigra\u003c/em\u003e (33.67\u0026thinsp;\u0026plusmn;\u0026thinsp;11.35) and lowest for \u003cem\u003eB. carinata\u003c/em\u003e \u0026times; \u003cem\u003eB. napus\u003c/em\u003e (18.68\u0026thinsp;\u0026plusmn;\u0026thinsp;6.62), whereas it was highest for \u003cem\u003eB. napus\u003c/em\u003e \u0026times; \u003cem\u003eB. carinata\u003c/em\u003e (33.06\u0026thinsp;\u0026plusmn;\u0026thinsp;13.41) in reciprocal crosses. \u003cem\u003eB. carinata\u003c/em\u003e \u0026times; \u003cem\u003eB. nigra\u003c/em\u003e had the highest mean pollen tube growth (21.23\u0026thinsp;\u0026plusmn;\u0026thinsp;5.33), followed by \u003cem\u003eB. carinata\u003c/em\u003e \u0026times; \u003cem\u003eB. napus\u003c/em\u003e (10.18\u0026thinsp;\u0026plusmn;\u0026thinsp;3.17), while \u003cem\u003eB. nigra \u0026times; B. carinata\u003c/em\u003e had the lowest mean growth among reciprocals (21.39\u0026thinsp;\u0026plusmn;\u0026thinsp;7.53). Pollen tube abnormalities such as coiling, bending, hairpin shapes, tube tip swelling, bifurcation, multiple tube emergence, and misdirected growth were observed. The highest incidence of abnormalities occurred when \u003cem\u003eB. carinata\u003c/em\u003e was used as a pollen parent with \u003cem\u003eB. napus\u003c/em\u003e. Further observations using fluorescent microscopy revealed pre-fertilization incompatibilities in the stylar and ovule regions. Crossability data revealed varying percentages of seed set (ranging from 0% in \u003cem\u003eB. nigra\u003c/em\u003e \u0026times; \u003cem\u003eB. carinata\u003c/em\u003e and \u003cem\u003eB. rapa\u003c/em\u003e \u0026times; \u003cem\u003eB. carinata\u003c/em\u003e to 25.64% in B. carinata \u0026times; B. nigra) and siliqua set (from 33% in \u003cem\u003eB. carinata\u003c/em\u003e \u0026times; \u003cem\u003eB. napus\u003c/em\u003e to 72% in selfed \u003cem\u003eB. carinata\u003c/em\u003e). Additionally, meiotic analysis revealed abnormalities such as improper alignment during M-I, M-II, A-I, A-II, and Telophase, indicating cytological instability in some hybrids. Correlation study further established the association among all of these traits. This study highlights critical reproductive and cytological barriers affecting hybrid success in \u003cem\u003eBrassica\u003c/em\u003e, offering valuable insights for breeders. By identifying suitable parent combinations and understanding hybridization limitations, it supports more efficient transfer of traits and the development of diverse, improved lines for future breeding programs.\u003c/p\u003e","manuscriptTitle":"Unravelling the secrets of wide hybridization among the brassica species","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-10-10 18:04:51","doi":"10.21203/rs.3.rs-7715506/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":"08c40529-4e80-4406-ba39-5156ac30b7b3","owner":[],"postedDate":"October 10th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-10-19T12:53:39+00:00","versionOfRecord":[],"versionCreatedAt":"2025-10-10 18:04:51","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7715506","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7715506","identity":"rs-7715506","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}