Optimizationof In-Vitro Regeneration Protocol for GI Tagged Aromatic Rice Landrace Chinnor

Optimizationof In-Vitro Regeneration Protocol for GI Tagged Aromatic Rice Landrace Chinnor | 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 Article Optimizationof In-Vitro Regeneration Protocol for GI Tagged Aromatic Rice Landrace Chinnor Poonam Kumari, Radheshyam Sharma, Shashank Bhargava, Stuti Sharma, and 6 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8876299/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 Chinnor, a traditional aromatic GI tag rice landrace, indigenous to Madhya Pradesh, is renowned for its superior grain quality and fragrance. However, its cultivation is limited to due to tall suture of the plant, occurrence of more insect and pest and less yield compared to commercial hybrids. Therefore, present study was undertaken to establish a reliable and efficient in-vitro plant regeneration protocol for chinnor land race for further improvement through gene editing and molecular breeding. To develop the in-vitro protocol, dehusked seeds were used as explant and inoculated in the basel MS medium. Callus induction was successfully achieved on Murashige and Skoog (MS) medium supplemented with 2.5 mg/l 2,4-D and 0.5 mg/L NAA, resulted a maximum callus induction rate of 90% after 28 days of culture. Subsequent shoot and root regeneration were optimized using a combination of auxins and cytokinins, with the highest regeneration rate (88%) observed on medium containing 2.0 mg/l Kinetin, 2.0 mg/l BAP, and 0.5 mg/l NAA. A stepwise hardening protocol was standardize to acclimatize the regenerated plantlets, with the cocopeat and farmyard manure (FYM)mixture (1:1) resulted the maximum survival rate of 86.67% under greenhouse conditions. This optimized protocol provides a robust platform for future biotechnological interventions through modification the traits of this GI tagged landrace. The findings significantly advance tissue culture methodologies for traditional rice genotypes and promote sustainable agricultural practices through the preservation of indigenous genetic resources. Biological sciences/Biological techniques Biological sciences/Biotechnology Biological sciences/Plant sciences Chinnor rice GI tag landrace In-vitro regeneration Callus induction Molecular breeding Tissue culture protocol Genetic fidelity 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-8876299","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":604936610,"identity":"e772bcce-b4cc-4cab-bdaa-2f9b7b5c7a80","order_by":0,"name":"Poonam Kumari","email":"","orcid":"","institution":"Biotechnology Centre, Jawaharlal Nehru Krishi Vishwa Vidyalaya (JNKVV)","correspondingAuthor":false,"prefix":"","firstName":"Poonam","middleName":"","lastName":"Kumari","suffix":""},{"id":604936611,"identity":"8414ad42-128b-4621-b92a-c829589730ad","order_by":1,"name":"Radheshyam Sharma","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABA0lEQVRIie3PMUsDMRTA8RyB3JLi+kqlfoWIkFOQ61fJEbipm6CZ5KBwXdr9/CKlY8uB09N+gIKcy00dztFBMFfBRYIZRfKfMuTHe4+QUOhvJklUQP9gpDPXlHBfYj+yqMLcm5AjoYOyJr+SZF6vmrf15XgSz9rXCndxwlECMS9p4SCnmN+dPyBccP6YiM7s6dVyYQneaBcBMpWjQQnZAqZsWOGeih3TIiqVdi0GJwc5+ujJ2YFZ++xBwE6JjlN4TzZUPM22jSWpm7S3w2V/C+bSLqapwHrbKFTKvZhewfv6fhzP6xY6k2qBWbHpjJq4yI++TrAjssKXfJ/gPyUUCoX+e59P31RQMJaUIAAAAABJRU5ErkJggg==","orcid":"","institution":"Biotechnology Centre, Jawaharlal Nehru Krishi Vishwa Vidyalaya (JNKVV)","correspondingAuthor":true,"prefix":"","firstName":"Radheshyam","middleName":"","lastName":"Sharma","suffix":""},{"id":604936612,"identity":"d6c4475c-e97d-4b98-b545-806bd83749e0","order_by":2,"name":"Shashank Bhargava","email":"","orcid":"","institution":"Biotechnology Centre, Jawaharlal Nehru Krishi Vishwa Vidyalaya (JNKVV)","correspondingAuthor":false,"prefix":"","firstName":"Shashank","middleName":"","lastName":"Bhargava","suffix":""},{"id":604936613,"identity":"4a263efd-0531-4c4b-a959-f6e12972215a","order_by":3,"name":"Stuti Sharma","email":"","orcid":"","institution":"Department of Genetics \u0026 Plant Breeding , College of Agriculture, Jawaharlal Nehru Krishi Vishwa Vidyalaya (JNKVV),","correspondingAuthor":false,"prefix":"","firstName":"Stuti","middleName":"","lastName":"Sharma","suffix":""},{"id":604936614,"identity":"d62d08d0-092c-46b6-a62e-fc0d4862f418","order_by":4,"name":"Ashish Kumar","email":"","orcid":"","institution":"Department of Genetics \u0026 Plant Breeding , College of Agriculture, Jawaharlal Nehru Krishi Vishwa Vidyalaya (JNKVV),","correspondingAuthor":false,"prefix":"","firstName":"Ashish","middleName":"","lastName":"Kumar","suffix":""},{"id":604936615,"identity":"a95261ce-1738-4587-9534-3e6ae4ce6a9d","order_by":5,"name":"Vikram Singh Gaur","email":"","orcid":"","institution":"College of Agriculture, Balaghat, Jawaharlal Nehru Krishi Vishwa Vidyalaya (JNKVV),","correspondingAuthor":false,"prefix":"","firstName":"Vikram","middleName":"Singh","lastName":"Gaur","suffix":""},{"id":604936616,"identity":"66d529e8-12cb-43f9-a854-a45ef72a99ea","order_by":6,"name":"Uttam Kumar Bisen","email":"","orcid":"","institution":"College of Agriculture, Balaghat, Jawaharlal Nehru Krishi Vishwa Vidyalaya (JNKVV),","correspondingAuthor":false,"prefix":"","firstName":"Uttam","middleName":"Kumar","lastName":"Bisen","suffix":""},{"id":604936617,"identity":"7460cfc8-abd5-48ae-b14d-d5f92f3e59a9","order_by":7,"name":"Soumya Patel","email":"","orcid":"","institution":"Department of Genetics \u0026 Plant Breeding , College of Agriculture, Jawaharlal Nehru Krishi Vishwa Vidyalaya (JNKVV),","correspondingAuthor":false,"prefix":"","firstName":"Soumya","middleName":"","lastName":"Patel","suffix":""},{"id":604936618,"identity":"8ca92306-5e0b-4e01-b869-6c510bbee892","order_by":8,"name":"Yogesh K. Ahlawat","email":"","orcid":"","institution":"Department of Life sciences, Faculty of Allied health sciences, SGT University","correspondingAuthor":false,"prefix":"","firstName":"Yogesh","middleName":"K.","lastName":"Ahlawat","suffix":""},{"id":604936619,"identity":"bb6ce654-6972-40d5-966b-d102cbdb4d32","order_by":9,"name":"Ihab Mohamed Moussa","email":"","orcid":"","institution":"King Saud University","correspondingAuthor":false,"prefix":"","firstName":"Ihab","middleName":"Mohamed","lastName":"Moussa","suffix":""}],"badges":[],"createdAt":"2026-02-14 02:24:17","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-8876299/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-8876299/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":105542636,"identity":"601b611f-1b55-4713-8357-35a81707a358","added_by":"auto","created_at":"2026-03-27 08:27:43","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":642749,"visible":true,"origin":"","legend":"","description":"","filename":"Chinnorrevised.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8876299/v1_covered_f9139f59-4cb1-47c5-b644-0942abc5e1c0.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"\u003cp\u003eOptimizationof In-Vitro Regeneration Protocol for GI Tagged Aromatic Rice Landrace Chinnor\u003c/p\u003e","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":"Chinnor rice, GI tag landrace, In-vitro regeneration, Callus induction, Molecular breeding, Tissue culture protocol, Genetic fidelity","lastPublishedDoi":"10.21203/rs.3.rs-8876299/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8876299/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eChinnor, a traditional aromatic GI tag rice landrace, indigenous to Madhya Pradesh, is \u0026nbsp;renowned for its superior grain quality and fragrance. However, its cultivation is limited to due \u0026nbsp;to tall suture of the plant, occurrence of more insect and pest and less yield compared to \u0026nbsp;commercial hybrids. Therefore, present study was undertaken to establish a reliable and efficient \u0026nbsp;in-vitro plant regeneration protocol for chinnor land race for further improvement through gene \u0026nbsp;editing and molecular breeding. To develop the in-vitro protocol, dehusked seeds were used as \u0026nbsp;explant and inoculated in the basel MS medium. Callus induction was successfully achieved on Murashige and Skoog (MS) medium supplemented with 2.5 mg/l 2,4-D and 0.5 mg/L NAA, resulted a maximum callus induction rate of 90% after 28 days of culture. Subsequent shoot and root regeneration were optimized using a combination of auxins and cytokinins, with the highest regeneration rate (88%) observed on medium containing 2.0 mg/l Kinetin, 2.0 mg/l BAP, and 0.5 \u0026nbsp;mg/l NAA. A stepwise hardening protocol was standardize to acclimatize the regenerated \u0026nbsp;plantlets, with the cocopeat and farmyard manure (FYM)mixture (1:1) resulted the maximum \u0026nbsp;survival rate of 86.67% under greenhouse conditions. This optimized protocol provides a robust platform for future biotechnological interventions through modification the traits of this GI \u0026nbsp;tagged landrace. The findings significantly advance tissue culture methodologies for traditional \u0026nbsp;rice genotypes and promote sustainable agricultural practices through the preservation of \u0026nbsp;indigenous genetic resources.\u003c/p\u003e","manuscriptTitle":"Optimizationof In-Vitro Regeneration Protocol for GI Tagged Aromatic Rice Landrace Chinnor","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-03-13 03:28:18","doi":"10.21203/rs.3.rs-8876299/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":"4a584827-fc7b-4a7f-81e9-92409683911a","owner":[],"postedDate":"March 13th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":64428734,"name":"Biological sciences/Biological techniques"},{"id":64428735,"name":"Biological sciences/Biotechnology"},{"id":64428736,"name":"Biological sciences/Plant sciences"}],"tags":[],"updatedAt":"2026-03-27T08:26:26+00:00","versionOfRecord":[],"versionCreatedAt":"2026-03-13 03:28:18","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8876299","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8876299","identity":"rs-8876299","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}