{"paper_id":"48cf7308-f5cc-472f-b4e7-402cd91e46f4","body_text":"Genome-wide characterization of WRKYs and function of SpWRKY40 and SpWRKY50 in Sesuvium portulacastrum | 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 characterization of WRKYs and function of SpWRKY40 and SpWRKY50 in Sesuvium portulacastrum Yaoxiu Li, Fengyan Fang, Xiaoyou Wu, Tongjing Cui, Yingyi Yu, and 4 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7513928/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 22 Dec, 2025 Read the published version in Functional & Integrative Genomics → Version 1 posted 18 You are reading this latest preprint version Abstract WRKYs represent a large family of plant transcription factors characterized by a highly conserved WRKY domain. WRKY transcription factors are important for plant growth, development, and responses to environmental stresses. However, this family has not been previously identified in Sesuvium portulacastrum . Here, we comprehensively identified 68 SpWRKYs from the genome of S. portulacastrum by phylogenetic relationships, chromosomal localization, collinearity analysis, gene structure, duplication events, and evolutionary pressure based on nonsynonymous substitution rate/synonymous substitution rate ratios. Promoter analysis revealed the presence of multiple cis -elements associated with abiotic stress signaling, developmental regulation, phytohormone responses, light responsiveness, and tissue-specific expression. We also characterized the physicochemical and structural properties of the SpWRKY -encoded proteins, including protein property of primary structure, secondary structures, and predicted three-dimensional models, subcellular localization, conserved motifs, and conserved domains. Furthermore, transcriptome profiling under cadmium, copper, and salt stress conditions demonstrated differential expression patterns of SpWRKYs . Among them, SpWRKY40 and SpWRKY51 showed significant indued under cadmium treatment and were further validated by quantitative real-time PCR. Subcellular localization and yeast one-hybrid assays both supporting SpWRKY40 and SpWRKY51 play as transcription factors. Our study provides a detailed overview of the SpWRKY family and their potential involvement in environment tolerance. The findings offer valuable candidate genes for future applications in improving stress tolerance in S. portulacastrum and related crop species. Sesuvium portulacastrum WRKY gene family Abiotic stress SpWRKY40 SpWRKY51 Transcription factor Full Text Additional Declarations No competing interests reported. Supplementary Files S1.png Fig. S1 Prediction of the tertiary structure of SpWRKYs. S2.png Fig. S2 The conserved motifs in SpWRKYs. TableS1.TranscriptomedataofSpWRKYsunderCdtreatment.xlsx TableS2.TranscriptomedataofSpWRKYsunderCutreatment.xlsx TableS3.TranscriptomedataofSpWRKYsundersaltstress.xlsx LiYXTable.docx Cite Share Download PDF Status: Published Journal Publication published 22 Dec, 2025 Read the published version in Functional & Integrative Genomics → Version 1 posted Editorial decision: Revision requested 25 Oct, 2025 Reviews received at journal 23 Oct, 2025 Reviews received at journal 22 Oct, 2025 Reviews received at journal 21 Oct, 2025 Reviews received at journal 12 Oct, 2025 Reviewers agreed at journal 09 Oct, 2025 Reviews received at journal 08 Oct, 2025 Reviewers agreed at journal 07 Oct, 2025 Reviewers agreed at journal 07 Oct, 2025 Reviewers agreed at journal 07 Oct, 2025 Reviewers agreed at journal 07 Oct, 2025 Reviewers agreed at journal 07 Oct, 2025 Reviewers agreed at journal 07 Oct, 2025 Reviewers agreed at journal 07 Oct, 2025 Reviewers invited by journal 07 Oct, 2025 Editor assigned by journal 12 Sep, 2025 Submission checks completed at journal 04 Sep, 2025 First submitted to journal 02 Sep, 2025 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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SpWRKY50 in Sesuvium portulacastrum\",\"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\":\"info@researchsquare.com\",\"identity\":\"functional-and-integrative-genomics\",\"isNatureJournal\":false,\"hasQc\":true,\"allowDirectSubmit\":false,\"externalIdentity\":\"fige\",\"sideBox\":\"Learn more about [Functional \\u0026 Integrative Genomics](http://link.springer.com/journal/10142)\",\"snPcode\":\"10142\",\"submissionUrl\":\"https://submission.nature.com/new-submission/10142/3\",\"title\":\"Functional \\u0026 Integrative Genomics\",\"twitterHandle\":\"\",\"acdcEnabled\":true,\"dfaEnabled\":true,\"editorialSystem\":\"stoa\",\"reportingPortfolio\":\"Springer Hybrid\",\"inReviewEnabled\":true,\"inReviewRevisionsEnabled\":false},\"keywords\":\"Sesuvium portulacastrum, WRKY gene family, Abiotic stress, SpWRKY40, SpWRKY51, Transcription factor\",\"lastPublishedDoi\":\"10.21203/rs.3.rs-7513928/v1\",\"lastPublishedDoiUrl\":\"https://doi.org/10.21203/rs.3.rs-7513928/v1\",\"license\":{\"name\":\"CC BY 4.0\",\"url\":\"https://creativecommons.org/licenses/by/4.0/\"},\"manuscriptAbstract\":\"\\u003cp\\u003eWRKYs represent a large family of plant transcription factors characterized by a highly conserved WRKY domain. WRKY transcription factors are important for plant growth, development, and responses to environmental stresses. However, this family has not been previously identified in \\u003cem\\u003eSesuvium portulacastrum\\u003c/em\\u003e. Here, we comprehensively identified 68 \\u003cem\\u003eSpWRKYs \\u003c/em\\u003efrom the genome of \\u003cem\\u003eS. portulacastrum\\u003c/em\\u003e by phylogenetic relationships, chromosomal localization, collinearity analysis, gene structure, duplication events, and evolutionary pressure based on nonsynonymous substitution rate/synonymous substitution rate ratios. Promoter analysis revealed the presence of multiple \\u003cem\\u003ecis\\u003c/em\\u003e-elements associated with abiotic stress signaling, developmental regulation, phytohormone responses, light responsiveness, and tissue-specific expression. We also characterized the physicochemical and structural properties of the \\u003cem\\u003eSpWRKY\\u003c/em\\u003e-encoded proteins, including protein property of primary structure, secondary structures, and predicted three-dimensional models, subcellular localization, conserved motifs, and conserved domains. Furthermore, transcriptome profiling under cadmium, copper, and salt stress conditions demonstrated differential expression patterns of \\u003cem\\u003eSpWRKYs\\u003c/em\\u003e. Among them, \\u003cem\\u003eSpWRKY40\\u003c/em\\u003e and \\u003cem\\u003eSpWRKY51\\u003c/em\\u003eshowed significant indued under cadmium treatment and were further validated by quantitative real-time PCR. Subcellular localization and yeast one-hybrid assays both supporting SpWRKY40 and SpWRKY51 play as transcription factors. Our study provides a detailed overview of the \\u003cem\\u003eSpWRKY\\u003c/em\\u003e family and their potential involvement in environment tolerance. The findings offer valuable candidate genes for future applications in improving stress tolerance in \\u003cem\\u003eS. portulacastrum\\u003c/em\\u003e and related crop species.\\u003c/p\\u003e\",\"manuscriptTitle\":\"Genome-wide characterization of WRKYs and function of SpWRKY40 and SpWRKY50 in Sesuvium portulacastrum\",\"msid\":\"\",\"msnumber\":\"\",\"nonDraftVersions\":[{\"code\":1,\"date\":\"2025-10-20 16:49:16\",\"doi\":\"10.21203/rs.3.rs-7513928/v1\",\"editorialEvents\":[{\"type\":\"communityComments\",\"content\":0},{\"type\":\"decision\",\"content\":\"Revision 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