Establishment of an efficient callus induction and in vitro plant regeneration protocol in okra (Abelmoschus esculentus(L.) Moench)

Establishment of an efficient callus induction and in vitro plant regeneration protocol in okra (Abelmoschus esculentus(L.) Moench) | 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 Establishment of an efficient callus induction and in vitro plant regeneration protocol in okra (Abelmoschus esculentus(L.) Moench) Manu A.N, Anshul Sharma, Ramesh Kumar Yadav, J. K. Ranjan, Krishan Kumar, and 7 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-9357079/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 7 You are reading this latest preprint version Abstract In the present investigation, a reproducible in vitro regeneration protocol was established for three okra cultivars— Pusa Sawani, Pusa Bhindi-5 and Pusa Lal Bhindi-1 -using hypocotyl and cotyledonary leaf explants. The explants were cultured on Murashige and Skoog medium (MS) comprising various auxin-cytokinin concentrations. MS mediumcontaining 2.0 mg/L 6-benzylaminopurine (BAP), 0.5 mg/L α-Naphthalene acetic acid (NAA) and 200 mg/L activated charcoal (AC) was the most responsive,showing a mean organogenic callus induction frequency of 73.79% from hypocotyl explants across all three cultivars. This was followed by MS medium having1.0 mg/LBAP, 0.5 mg/L NAA,200 mg/L AC (66.50%) and 2.0 mg/L Zeatin,0.2 mg/L NAA,200 mg/LAC (65.29%). Cotyledonary leaf explants exhibiteda maximum callus induction frequency of 66.73% under the same hormonal combination. The lowest callus induction (23.18–24.28%) was observed on MS medium containing1.0 TDZ, 0.1 NAA,200 AC for both theexplants.The highest shoot induction (71.17%) with the earliest shoot bud emergence (13.25 days) was achieved on MS medium augmented with 2.0mg/L BAP, 0.5 mg/L NAA, and 200 mg/L AC. Optimal rooting was obtained on MS medium containing 0.2 mg/L NAA and 200 mg/L AC, resulting in 91.14% rooting, the earliest root initiation (14.82 days), and the longest roots (4.96 cm) in cultivar Pusa Lal Bhindi-1. As per our information, this is the first report detailing a robust and reproducible in vitro callus induction and regeneration protocol for the okra cultivars Pusa Sawani, Pusa Bhindi-5 and Pusa Lal Bhindi-1. Callus induction Hypocotyl Cotyledon Somatic Embryogenesis Regeneration Okra Abelmoschus esculentus Full Text Additional Declarations No competing interests reported. Cite Share Download PDF Status: Under Review Version 1 posted Reviewers agreed at journal 09 May, 2026 Reviewers agreed at journal 05 May, 2026 Reviewers invited by journal 04 May, 2026 Editor assigned by journal 29 Apr, 2026 Editor invited by journal 17 Apr, 2026 Submission checks completed at journal 17 Apr, 2026 First submitted to journal 17 Apr, 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. We do this by developing innovative software and high quality services for the global research community. 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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-9357079","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":632086749,"identity":"76d0d7ab-520b-4db1-a71e-37342a1d8b30","order_by":0,"name":"Manu A.N","email":"","orcid":"","institution":"Indian Agricultural Research Institute","correspondingAuthor":false,"prefix":"","firstName":"Manu","middleName":"","lastName":"A.N","suffix":""},{"id":632086750,"identity":"4208b0ab-b3e8-439a-bbbe-a3e8d804583b","order_by":1,"name":"Anshul Sharma","email":"","orcid":"","institution":"Indian Agricultural Research Institute","correspondingAuthor":false,"prefix":"","firstName":"Anshul","middleName":"","lastName":"Sharma","suffix":""},{"id":632086751,"identity":"e1ced08b-e706-4436-928b-4c47404b3ac4","order_by":2,"name":"Ramesh Kumar Yadav","email":"","orcid":"","institution":"Indian Agricultural Research Institute","correspondingAuthor":false,"prefix":"","firstName":"Ramesh","middleName":"Kumar","lastName":"Yadav","suffix":""},{"id":632086752,"identity":"cf346cc5-f0ad-437d-b100-63c3ebf2562a","order_by":3,"name":"J. 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The explants were cultured on Murashige and Skoog medium (MS) comprising various auxin-cytokinin concentrations. MS mediumcontaining 2.0 mg/L 6-benzylaminopurine (BAP), 0.5 mg/L α-Naphthalene acetic acid (NAA) and 200 mg/L activated charcoal (AC) was the most responsive,showing a mean organogenic callus induction frequency of 73.79% from hypocotyl explants across all three cultivars. This was followed by MS medium having1.0 mg/LBAP, 0.5 mg/L NAA,200 mg/L AC (66.50%) and 2.0 mg/L Zeatin,0.2 mg/L NAA,200 mg/LAC (65.29%). Cotyledonary leaf explants exhibiteda maximum callus induction frequency of 66.73% under the same hormonal combination. The lowest callus induction (23.18\u0026ndash;24.28%) was observed on MS medium containing1.0 TDZ, 0.1 NAA,200 AC for both theexplants.The highest shoot induction (71.17%) with the earliest shoot bud emergence (13.25 days) was achieved on MS medium augmented with 2.0mg/L BAP, 0.5 mg/L NAA, and 200 mg/L AC. 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