Genome-Wide Identification of GRF and GIF Genes in Larix kaempferi and Their Potential Association with Somatic Embryogenesis, Hormone Responses, and miR396-Mediated Regulation

Genome-Wide Identification of GRF and GIF Genes in Larix kaempferi and Their Potential Association with Somatic Embryogenesis, Hormone Responses, and miR396-Mediated Regulation | 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 Genome-Wide Identification of GRF and GIF Genes in Larix kaempferi and Their Potential Association with Somatic Embryogenesis, Hormone Responses, and miR396-Mediated Regulation Yuqin Huang, Ruxin Zhang, Xin Li, Liwang Qi, Lifeng Zhang This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-9181763/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 14 You are reading this latest preprint version Abstract This study systematically identified and characterized the Growth-regulating factors ( GRFs ) and GRF-interacting factors ( GIFs ) gene families in Larix kaempferi , revealing five LkaGRF and three LkaGIF genes. All LkaGRF proteins contain conserved QLQ and WRC domains, while LkaGIF proteins possess the SSXT domain, confirming their structural conservation across land plants. Phylogenetic analysis placed most LkaGRFs in a gymnosperm-specific clade, indicating ancient lineage diversification, whereas LkaGIFs showed both conserved and gymnosperm-specific branches. Predicted miR396 target sites were detected in all LkaGRF transcripts, and 5′ RLM-RACE further supported miR396-directed cleavage of LkaGRF3 and LkaGRF5. Promoter analysis revealed abundant hormone- and stress-responsive cis-elements. Expression profiling demonstrated that both gene families are significantly upregulated at day 28 during somatic embryogenesis, corresponding to the critical stage of cotyledon differentiation. Hormonal treatments showed that most LkaGRFs are rapidly downregulated by IAA, ABA, GA, and SA, LkaGRF1 showed a generally sustained repressed expression pattern under all five hormone treatments, whereas LkaGRF3 and LkaGRF5 were continuously and significantly downregulated by IAA, ABA, GA, and SA. These results support the potential involvement of the miR396-GRF/GIF module in somatic embryogenesis and hormone-responsive developmental processes in L. kaempferi , and provide candidate regulators for future functional studies in conifer embryogenic systems. Growth-Regulating Factor (GRF) GRF-Interacting Factor (GIF) Larix kaempferi Somatic Embryogenesis miR396 Full Text Additional Declarations No competing interests reported. Supplementary Files supplementaryfile.xlsx Cite Share Download PDF Status: Under Review Version 1 posted Editorial decision: Revision requested 26 Apr, 2026 Reviews received at journal 24 Apr, 2026 Reviews received at journal 21 Apr, 2026 Reviews received at journal 10 Apr, 2026 Reviews received at journal 06 Apr, 2026 Reviewers agreed at journal 03 Apr, 2026 Reviewers agreed at journal 02 Apr, 2026 Reviewers agreed at journal 01 Apr, 2026 Reviewers agreed at journal 01 Apr, 2026 Reviewers agreed at journal 28 Mar, 2026 Reviewers invited by journal 27 Mar, 2026 Editor assigned by journal 25 Mar, 2026 Submission checks completed at journal 22 Mar, 2026 First submitted to journal 20 Mar, 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. 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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-9181763","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":614064025,"identity":"c69eeb1e-2974-491a-a7a2-35e090067318","order_by":0,"name":"Yuqin Huang","email":"","orcid":"","institution":"Chinese Academy of Forestry","correspondingAuthor":false,"prefix":"","firstName":"Yuqin","middleName":"","lastName":"Huang","suffix":""},{"id":614064026,"identity":"dd7b25a0-5ce5-47ff-af0c-601dc858f4ba","order_by":1,"name":"Ruxin Zhang","email":"","orcid":"","institution":"Hebei Agricultural 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