Revealing the Response Mechanism of Winter Rapeseed (Brassica rapa L. subsp. oleifera ) to Freezing Stress Based on Transcriptome Analysis 

Revealing the Response Mechanism of Winter Rapeseed (Brassica rapa L. subsp. oleifera ) to Freezing Stress Based on Transcriptome Analysis | 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 Revealing the Response Mechanism of Winter Rapeseed (Brassica rapa L. subsp. oleifera ) to Freezing Stress Based on Transcriptome Analysis Xinxin Li, Juncheng Zhu, Yuanyuan Pu, Li Ma, Lijun Liu, Wangtian Wang, and 4 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7920671/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 9 You are reading this latest preprint version Abstract Winter rapeseed ( Brassica rapa L. subsp.oleifera ) is an important oilseed crop in northern China, where low-temperature stress imposes severe constraints on its yield. Its leaves dry up and wither during the overwintering period, but its roots stay deeply embedded in the soil. Following March of the next year, new leaves grow from the growing point and it enters reproductive growth. Therefore, we carried out this research for exploring the regulatory mechanisms and physiological-biochemical responses of roots in different rapeseed varieties as the temperature decrease before wintering. This study used a strong cold-tolerant variety "Longyou 7 (L7)" and a moderate cold-tolerant variety "Longyou 99 (L99)" as experimental materials. The materials were planted in field to analyze the effects of natural low-temperature stress on rapeseed growth, physiological characteristics, hormone levels, and transcriptional levels before wintering. Additionally, transcriptome sequencing (RNA-seq) was combined to decipher the molecular regulatory mechanisms. Results showed that as temperature decrease, contents of proline (Pro), soluble sugar (SS), soluble protein (SP), and salicylic acid (SA) in roots of the strong cold-tolerant variety L7 were all significantly higher than those in L99, while contents of malondialdehyde (MDA) and gibberellin (GA₃) in L7 were significantly lower than those in L99. RNA-seq results revealed that differentially expressed genes (DEGs) were significantly involved in phenylpropane biosynthesis, carbon metabolism, starch and sucrose metabolism, MAPK signaling pathway, ribosome, proteasome, and protein processing. Among these, the differences in the expression of genes related to cell signal transduction (MAPK signaling pathway) and metabolism were particularly prominent. Through weighted gene co-expression network analysis (WGCNA), 9 candidate genes related to protein kinases, plant hormones, transcription factors, and signal transduction were identified in the MAPK and 2 other modules. winter rapeseed freezing stress root transcriptome differentially expressed genes Full Text Additional Declarations No competing interests reported. Cite Share Download PDF Status: Under Review Version 1 posted Editorial decision: Revision requested 05 Dec, 2025 Reviews received at journal 02 Dec, 2025 Reviews received at journal 28 Nov, 2025 Reviewers agreed at journal 16 Nov, 2025 Reviewers agreed at journal 06 Nov, 2025 Reviewers invited by journal 06 Nov, 2025 Editor assigned by journal 05 Nov, 2025 Submission checks completed at journal 04 Nov, 2025 First submitted to journal 31 Oct, 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. 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-7920671","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":545767432,"identity":"e5dc4c76-2e09-4fc7-b89e-48fe1b7064dd","order_by":0,"name":"Xinxin Li","email":"","orcid":"","institution":"Gansu Agricultural University","correspondingAuthor":false,"prefix":"","firstName":"Xinxin","middleName":"","lastName":"Li","suffix":""},{"id":545767433,"identity":"784a8294-999f-4f18-862b-21da293d6547","order_by":1,"name":"Juncheng Zhu","email":"","orcid":"","institution":"Gansu Agricultural 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","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":"chemical-and-biological-technologies-in-agriculture","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"Learn more about [Chemical and Biological Technologies in Agriculture](https://chembioagro.springeropen.com/)","snPcode":"40538","submissionUrl":"https://submission.nature.com/new-submission/40538/3","title":"Chemical and Biological Technologies in 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Its leaves dry up and wither during the overwintering period, but its roots stay deeply embedded in the soil. Following March of the next year, new leaves grow from the growing point and it enters reproductive growth. Therefore, we carried out this research for exploring the regulatory mechanisms and physiological-biochemical responses of roots in different rapeseed varieties as the temperature decrease before wintering. This study used a strong cold-tolerant variety \"Longyou 7 (L7)\" and a moderate cold-tolerant variety \"Longyou 99 (L99)\" as experimental materials. The materials were planted in field to analyze the effects of natural low-temperature stress on rapeseed growth, physiological characteristics, hormone levels, and transcriptional levels before wintering. Additionally, transcriptome sequencing (RNA-seq) was combined to decipher the molecular regulatory mechanisms. Results showed that as temperature decrease, contents of proline (Pro), soluble sugar (SS), soluble protein (SP), and salicylic acid (SA) in roots of the strong cold-tolerant variety L7 were all significantly higher than those in L99, while contents of malondialdehyde (MDA) and gibberellin (GA₃) in L7 were significantly lower than those in L99. RNA-seq results revealed that differentially expressed genes (DEGs) were significantly involved in phenylpropane biosynthesis, carbon metabolism, starch and sucrose metabolism, MAPK signaling pathway, ribosome, proteasome, and protein processing. Among these, the differences in the expression of genes related to cell signal transduction (MAPK signaling pathway) and metabolism were particularly prominent. Through weighted gene co-expression network analysis (WGCNA), 9 candidate genes related to protein kinases, plant hormones, transcription factors, and signal transduction were identified in the MAPK and 2 other modules.\u003c/p\u003e","manuscriptTitle":"Revealing the Response Mechanism of Winter Rapeseed (Brassica rapa L. subsp. oleifera ) to Freezing Stress Based on Transcriptome Analysis ","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-11-17 03:44:33","doi":"10.21203/rs.3.rs-7920671/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-12-05T09:10:05+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-12-03T03:01:33+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-11-29T02:02:30+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"73386312785402326422216243772182883502","date":"2025-11-17T02:51:57+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"77185301610574988889737821910575311920","date":"2025-11-07T01:04:37+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-11-06T11:53:47+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-11-05T07:19:16+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-11-04T11:28:11+00:00","index":"","fulltext":""},{"type":"submitted","content":"Chemical and Biological Technologies in Agriculture","date":"2025-10-31T07:45:06+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"chemical-and-biological-technologies-in-agriculture","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"","sideBox":"Learn more about [Chemical and Biological Technologies in Agriculture](https://chembioagro.springeropen.com/)","snPcode":"40538","submissionUrl":"https://submission.nature.com/new-submission/40538/3","title":"Chemical and Biological Technologies in Agriculture","twitterHandle":"@SpringerPlants","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"44cac08e-44de-4d62-8057-e116cd7f26ba","owner":[],"postedDate":"November 17th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[],"tags":[],"updatedAt":"2026-03-21T16:08:30+00:00","versionOfRecord":[],"versionCreatedAt":"2025-11-17 03:44:33","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7920671","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7920671","identity":"rs-7920671","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}