Concentration-dependent effects of melatonin: physiological and metabolic processes in Pfaffia glomerata plants grown in vitro

Concentration-dependent effects of melatonin: physiological and metabolic processes in Pfaffia glomerata plants grown in vitro | 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 Concentration-dependent effects of melatonin: physiological and metabolic processes in Pfaffia glomerata plants grown in vitro José V. S. Silva, Tatiane D. Silva, Sérgio H. S. Felipe, Diego S. Batista, and 7 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5271110/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 01 May, 2025 Read the published version in Plant Cell, Tissue and Organ Culture (PCTOC) → Version 1 posted 5 You are reading this latest preprint version Abstract Melatonin is a multifunctional molecule active in various metabolic processes, influencing the accumulation of bioactive metabolites in medicinal plants such as Pfaffia glomerata. This species has pharmacological and medicinal properties, mainly due to the 20-hydroxyecdysone (20-E) production. This study aimed to evaluate the effect of exogenous melatonin on the growth and development, physiology, and content of 20-E in P. glomerata plants. To achieve this, P. glomerata was grown in vitro for 35 days in a culture medium supplemented with 0, 10, 100, 500, and 1000 μM melatonin. Morphophysiological parameters, antioxidant defenses, and 20-E production were subsequently assessed. Melatonin reduced pigment content, with higher concentrations (≥ 500 µM) impairing plant growth and biomass accumulation. However, melatonin also triggered antioxidant response mechanisms, leading to increased antioxidant enzymes activity and flavonoid contents, while simultaneously reducing lipid peroxidation in leaves and H2O2 accumulation in roots. Moreover, melatonin increased the levels of amino acids, proline and proteins in the roots, while also promoting the production of 20-E (at 10 µM) and increasing the contents of zeatin, ACC, and methyl jasmonate, which are important stress-related hormones. Our findings provide further insight into the role of melatonin as a regulatory molecule in plants, with its effects being concentration-dependent. Brazilian-ginseng 20-hydroxyecdysone hydrogen peroxide antidoxidant activity Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Full Text Cite Share Download PDF Status: Published Journal Publication published 01 May, 2025 Read the published version in Plant Cell, Tissue and Organ Culture (PCTOC) → Version 1 posted Editorial decision: Major revisions 16 Feb, 2025 Reviewers agreed at journal 30 Oct, 2024 Reviewers invited by journal 30 Oct, 2024 Editor assigned by journal 17 Oct, 2024 First submitted to journal 15 Oct, 2024 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-5271110","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":372034462,"identity":"3372bc3d-9887-40da-a08f-e3d86d9630f3","order_by":0,"name":"José V. 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Batista","email":"","orcid":"","institution":"Paraiba Federal University: Universidade Federal da Paraiba","correspondingAuthor":false,"prefix":"","firstName":"Diego","middleName":"S.","lastName":"Batista","suffix":""},{"id":372034466,"identity":"35a89fb4-a63e-4156-9ba8-db6766b90be1","order_by":4,"name":"Juliane M. Henschel","email":"","orcid":"","institution":"Paraiba Federal University: Universidade Federal da Paraiba","correspondingAuthor":false,"prefix":"","firstName":"Juliane","middleName":"M.","lastName":"Henschel","suffix":""},{"id":372034467,"identity":"bd66f4a9-0bf8-47c1-b768-6cb004cdaead","order_by":5,"name":"Evandro A. Fortini","email":"","orcid":"","institution":"North Dakota State University","correspondingAuthor":false,"prefix":"","firstName":"Evandro","middleName":"A.","lastName":"Fortini","suffix":""},{"id":372034468,"identity":"4e33aa17-6496-48d4-933f-3f033a3caed7","order_by":6,"name":"Letícia M. Farias","email":"","orcid":"","institution":"Federal University of Vicosa: Universidade Federal de Vicosa","correspondingAuthor":false,"prefix":"","firstName":"Letícia","middleName":"M.","lastName":"Farias","suffix":""},{"id":372034469,"identity":"87fd91ed-a31e-4899-86b1-d931b34106df","order_by":7,"name":"João P. V. Leite","email":"","orcid":"","institution":"Federal University of Vicosa: Universidade Federal de Vicosa","correspondingAuthor":false,"prefix":"","firstName":"João","middleName":"P. V.","lastName":"Leite","suffix":""},{"id":372034470,"identity":"8c0da288-2e2e-4a19-9191-3061e894cbae","order_by":8,"name":"Michelle M. V. Almeida","email":"","orcid":"","institution":"Federal University of Vicosa: Universidade Federal de Vicosa","correspondingAuthor":false,"prefix":"","firstName":"Michelle","middleName":"M. V.","lastName":"Almeida","suffix":""},{"id":372034471,"identity":"a883e395-bf1e-45ae-83fa-b17cf40236ea","order_by":9,"name":"Kleiton L. G. Machado","email":"data:image/png;base64,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","orcid":"https://orcid.org/0000-0002-7935-4439","institution":"UFV: Universidade Federal de Vicosa","correspondingAuthor":true,"prefix":"","firstName":"Kleiton","middleName":"L. G.","lastName":"Machado","suffix":""},{"id":372034472,"identity":"51786ede-06b9-4388-a8ff-e3355e9053ec","order_by":10,"name":"Wagner C. Otoni","email":"","orcid":"","institution":"Federal University of Vicosa: Universidade Federal de Vicosa","correspondingAuthor":false,"prefix":"","firstName":"Wagner","middleName":"C.","lastName":"Otoni","suffix":""}],"badges":[],"createdAt":"2024-10-15 19:57:21","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-5271110/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-5271110/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s11240-025-03071-w","type":"published","date":"2025-05-01T15:57:42+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":68680967,"identity":"0e80252f-bd04-47db-a014-cf668d610db4","added_by":"auto","created_at":"2024-11-11 03:38:07","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":230559,"visible":true,"origin":"","legend":"\u003cp\u003eBiometric assessments of Pfaffia glomerata grown 35 days under different \u0026nbsp;melatonin concentrations (from 0 to 1000 μM). Bar: 5 cm. Vertical bars represent \u0026nbsp;standard errors. A. Plant stem and leaves architecture, and roots growth pattern; B. Stem \u0026nbsp;length; C. Root length; D. Leaf dry weight; E. Stem dry weight; F. Root dry weight; G. \u0026nbsp;Number of leaves; H. Leaf area; Groups sharing the same letters are deemed statistically \u0026nbsp;homogeneous, as established by Duncan's test (P ≤ 0.05). Bars represent the \u0026nbsp;mean ± standard error (n = 4).\u003c/p\u003e","description":"","filename":"Figure1.png","url":"https://assets-eu.researchsquare.com/files/rs-5271110/v1/8526718b174348ae91d2b509.png"},{"id":68681767,"identity":"a883e3e8-c5fe-4067-990d-d0f0e941546d","added_by":"auto","created_at":"2024-11-11 03:46:06","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":85936,"visible":true,"origin":"","legend":"\u003cp\u003ePigment quantification on P. glomerata plants submitted to different melatonin \u0026nbsp;concentrations. A. Chlorophyll a, B. Chlorophyll b, C. Chlorophyll a/b ratio, D. \u0026nbsp;Carotenoids, E. Anthocyanin content. Bars represent the mean ± standard error (n = 4). \u0026nbsp;Groups sharing the same letters are deemed statistically homogeneous, as established by \u0026nbsp;Duncan's test (P ≤ 0.05).\u003c/p\u003e","description":"","filename":"Figure2.png","url":"https://assets-eu.researchsquare.com/files/rs-5271110/v1/c53079ae012028d1549fa42c.png"},{"id":68680963,"identity":"b65889ae-6714-40df-90c5-07ba1894185f","added_by":"auto","created_at":"2024-11-11 03:38:06","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":110090,"visible":true,"origin":"","legend":"\u003cp\u003eSoluble sugar and starch quantity on leaves and roots of P. glomerata plants \u0026nbsp;grown under different melatonin concentrations. A. Soluble sugars and B. Starch contents \u0026nbsp;on leaves; C. Soluble sugars and D. Starch levels on roots. Bars represent the \u0026nbsp;mean ± standard error (n = 4). Groups sharing the same letters are deemed statistically \u0026nbsp;homogeneous, as established by Duncan's test (P ≤ 0.05).\u003c/p\u003e","description":"","filename":"Figure3.png","url":"https://assets-eu.researchsquare.com/files/rs-5271110/v1/c4433485081bb95bc8e2cecc.png"},{"id":68680966,"identity":"f71f4981-0e86-48f2-9691-fe927fb6b826","added_by":"auto","created_at":"2024-11-11 03:38:07","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":81786,"visible":true,"origin":"","legend":"\u003cp\u003eProtein metabolism in P. glomerata plants grown under different melatonin \u0026nbsp;concentrations. A. Protein, B. Aminoacids, and C. Proline content in leaves; D. Protein, \u0026nbsp;E. Aminoacids, and F. Proline levels in roots. Bars represent the mean ± standard error (n \u0026nbsp;= 4). Groups sharing the same letters are deemed statistically homogeneous, as \u0026nbsp;established by Duncan's test (P ≤ 0.05).\u003c/p\u003e","description":"","filename":"Figure4.png","url":"https://assets-eu.researchsquare.com/files/rs-5271110/v1/eef5f195b78b80da08a23513.png"},{"id":68680971,"identity":"420d0eed-a46b-49be-b159-0ad80fe636fd","added_by":"auto","created_at":"2024-11-11 03:38:08","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":83430,"visible":true,"origin":"","legend":"\u003cp\u003eOxidative metabolism enzymes activity in P. glomerata plants submitted to \u0026nbsp;melatonin concentrations. A. Catalase (CAT), B. Ascorbate Peroxidase (APX), C. \u0026nbsp;Peroxidase Oxidoreductase (POD), and D. Superoxide Dismutase (SOD) activities in \u0026nbsp;leaves; E. CAT, F. APX, G. POD, and H. SOD activities in roots. Bars represent the \u0026nbsp;mean ± standard error (n = 4). Groups sharing the same letters are deemed statistically \u0026nbsp;homogeneous, as established by Duncan's test (P ≤ 0.05).\u003c/p\u003e","description":"","filename":"Figure5.png","url":"https://assets-eu.researchsquare.com/files/rs-5271110/v1/e11df85492c01627a5b67dfb.png"},{"id":68680965,"identity":"0be73604-72ee-4b3c-b864-1e642f3f0ac3","added_by":"auto","created_at":"2024-11-11 03:38:07","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":84066,"visible":true,"origin":"","legend":"\u003cp\u003ePlant stress-related metabolites on 615 P. glomerata leaves and roots grown under \u0026nbsp;different melatonin concentrations. A. Malondialdehyde (MDA), B. Free Hydrogen \u0026nbsp;Peroxide (H2O2), C. Phenols and D. Flavonoids levels in leaves; E. MDA, F. Free H2O2, \u0026nbsp;G. Phenols, and H. Flavonoids content in roots. Bars represent the mean ± standard error \u0026nbsp;(n = 4). Groups sharing the same letters are deemed statistically homogeneous, as \u0026nbsp;established by Duncan's test (P ≤ 0.05).\u003c/p\u003e","description":"","filename":"Figure6.png","url":"https://assets-eu.researchsquare.com/files/rs-5271110/v1/5f74ec166f045d0f6cbd693e.png"},{"id":68680968,"identity":"006d9b5d-4ea9-49d8-b6cb-144113bbd53d","added_by":"auto","created_at":"2024-11-11 03:38:08","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":65357,"visible":true,"origin":"","legend":"\u003cp\u003ePhytohormones and 20-hydroxyecdysone content in Pfaffia glomerata leaves \u0026nbsp;after 35 days of in vitro cultivation, under different concentrations of melatonin (0; 10; \u0026nbsp;100; 500 and 1000 μM). A. Abscisic acid (ABA); B. Indole-3-acetic acid (IAA); C. 1- \u0026nbsp;aminocyclopropane-1-carboxylic acid (ACC); D. zeatin (Zea); E. Methyl jasmonate \u0026nbsp;(MeJa); F. Gibberellic acid (GA3); G. Brassinosteroids (BR); H. 20-hydroxyecdysone \u0026nbsp;(20-E). Bars represent the mean ± standard error (n = 4). Groups sharing the same letters \u0026nbsp;are deemed statistically homogeneous, as established by Duncan's test (P ≤ 0.05).\u003c/p\u003e","description":"","filename":"Figure7.png","url":"https://assets-eu.researchsquare.com/files/rs-5271110/v1/d514ff401ca3cb5a22dfd4dd.png"},{"id":81987809,"identity":"659268a9-d8c8-4b7a-9996-7424b963f7ac","added_by":"auto","created_at":"2025-05-05 16:06:11","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":968376,"visible":true,"origin":"","legend":"","description":"","filename":"PglomerataPCTOCFINAL2024.pdf","url":"https://assets-eu.researchsquare.com/files/rs-5271110/v1_covered_0a1df59d-414f-48bf-80ff-5e13b6a761a8.pdf"}],"financialInterests":"","formattedTitle":"Concentration-dependent effects of melatonin: physiological and metabolic processes in Pfaffia glomerata plants grown in vitro","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":"plant-cell-tissue-and-organ-culture-pctoc","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"pcto","sideBox":"Learn more about [Plant Cell, Tissue and Organ Culture (PCTOC)](https://www.springer.com/journal/11240)","snPcode":"11240","submissionUrl":"https://submission.nature.com/new-submission/11240/3","title":"Plant Cell, Tissue and Organ Culture (PCTOC)","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Brazilian-ginseng, 20-hydroxyecdysone, hydrogen peroxide, antidoxidant activity","lastPublishedDoi":"10.21203/rs.3.rs-5271110/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5271110/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"Melatonin is a multifunctional molecule active in various metabolic processes, influencing the accumulation of bioactive metabolites in medicinal plants such as Pfaffia glomerata. This species has pharmacological and medicinal properties, mainly due to the 20-hydroxyecdysone (20-E) production. This study aimed to evaluate the effect of exogenous melatonin on the growth and development, physiology, and content of 20-E in P. glomerata plants. To achieve this, P. glomerata was grown in vitro for 35 days in a culture medium supplemented with 0, 10, 100, 500, and 1000 μM melatonin. Morphophysiological parameters, antioxidant defenses, and 20-E production were subsequently assessed. Melatonin reduced pigment content, with higher concentrations (≥ 500 µM) impairing plant growth and biomass accumulation. However, melatonin also triggered antioxidant response mechanisms, leading to increased antioxidant enzymes activity and flavonoid contents, while simultaneously reducing lipid peroxidation in leaves and H2O2 accumulation in roots. Moreover, melatonin increased the levels of amino acids, proline and proteins in the roots, while also promoting the production of 20-E (at 10 µM) and increasing the contents of zeatin, ACC, and methyl jasmonate, which are important stress-related hormones. Our findings provide further insight into the role of melatonin as a regulatory molecule in plants, with its effects being concentration-dependent.","manuscriptTitle":"Concentration-dependent effects of melatonin: physiological and metabolic processes in Pfaffia glomerata plants grown in vitro","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-11-11 03:38:02","doi":"10.21203/rs.3.rs-5271110/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Major revisions","date":"2025-02-17T02:59:22+00:00","index":"","fulltext":""},{"type":"reviewerAgreed","content":"","date":"2024-10-30T05:30:20+00:00","index":0,"fulltext":""},{"type":"reviewersInvited","content":"","date":"2024-10-30T05:14:56+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-10-17T09:23:45+00:00","index":"","fulltext":""},{"type":"submitted","content":"Plant Cell, Tissue and Organ Culture (PCTOC)","date":"2024-10-15T15:57:00+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"plant-cell-tissue-and-organ-culture-pctoc","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"pcto","sideBox":"Learn more about [Plant Cell, Tissue and Organ Culture (PCTOC)](https://www.springer.com/journal/11240)","snPcode":"11240","submissionUrl":"https://submission.nature.com/new-submission/11240/3","title":"Plant Cell, Tissue and Organ Culture (PCTOC)","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"2aac5ecf-d746-4157-a857-60cac53a91b6","owner":[],"postedDate":"November 11th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2025-05-05T16:01:45+00:00","versionOfRecord":{"articleIdentity":"rs-5271110","link":"https://doi.org/10.1007/s11240-025-03071-w","journal":{"identity":"plant-cell-tissue-and-organ-culture-pctoc","isVorOnly":false,"title":"Plant Cell, Tissue and Organ Culture (PCTOC)"},"publishedOn":"2025-05-01 15:57:42","publishedOnDateReadable":"May 1st, 2025"},"versionCreatedAt":"2024-11-11 03:38:02","video":"","vorDoi":"10.1007/s11240-025-03071-w","vorDoiUrl":"https://doi.org/10.1007/s11240-025-03071-w","workflowStages":[]},"version":"v1","identity":"rs-5271110","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5271110","identity":"rs-5271110","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}