Evolutionary and Functional Insights into PSI-Containing Aspartic Proteinases in Tomato

Evolutionary and Functional Insights into PSI-Containing Aspartic Proteinases in Tomato | 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 Evolutionary and Functional Insights into PSI-Containing Aspartic Proteinases in Tomato Miguel Sampaio, João Neves, João Monteiro, Cristiano Soares, Fernanda Fidalgo, and 4 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7245797/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Aspartic proteinases (APs) are essential proteolytic enzymes involved in plant growth, development, and defence. This study investigates PSI-containing Aspartic Proteinases (APs) in Solanum lycopersicum (tomato plant), focusing on their molecular characteristics, tissue-specific expression, and response to abiotic stress. While 58 APs were identified in S. lycopersicum , only five (AP V, AP W, AP X, AP Y, and AP Z) contained a C-terminal vacuolar sorting domain and the Plant Specific Insert (PSI) domain, a unique domain implicated in vacuolar trafficking and stress responses. These PSI-containing APs were further analyzed through phylogenetics, revealing their evolutionary relationships with homologous APs from other species. Expression profiling showed that most PSI-containing APs are highly expressed in cotyledons and roots, with AP Y being distinctively predominant in reproductive tissues. Under osmotic and saline stress, expression patterns varied, with AP Z exhibiting a broader stress sensitivity. Biochemical analyses confirmed that severe osmotic stress led to increased oxidative stress markers and antioxidant responses, suggestive of activation of the plant-adaptive mechanisms to abiotic stress. As a whole, it is shown that PSI-containing APs from Solanum lycopersicum are strictly regulated within the plant and exhibit varied responses to environmental stressors. Aspartic proteases Plant specific insert endomembrane trafficking Solanum lycopersicum. Full Text Additional Declarations No competing interests reported. Supplementary Files SupplementarytablesSampaioetal.docx Cite Share Download PDF Status: Posted Version 1 posted 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. 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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-7245797","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":498495425,"identity":"140bd4be-53f1-41ac-ba48-bd012632c682","order_by":0,"name":"Miguel Sampaio","email":"","orcid":"","institution":"GreenUPorto – Sustainable Agrifood Production Research Centre and INOV4AGRO - Institute for Innovation, Training and Sustainability of Agrifood Production, University of Porto","correspondingAuthor":false,"prefix":"","firstName":"Miguel","middleName":"","lastName":"Sampaio","suffix":""},{"id":498495426,"identity":"64272a3a-a038-4789-87c9-ec9124f5824e","order_by":1,"name":"João Neves","email":"","orcid":"","institution":"ICBAS, University of Porto","correspondingAuthor":false,"prefix":"","firstName":"João","middleName":"","lastName":"Neves","suffix":""},{"id":498495427,"identity":"589a868b-b54e-4e3c-a002-58b74f16cee5","order_by":2,"name":"João Monteiro","email":"","orcid":"","institution":"GreenUPorto – Sustainable Agrifood Production Research Centre and INOV4AGRO - 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This study investigates PSI-containing Aspartic Proteinases (APs) in \u003cem\u003eSolanum lycopersicum\u003c/em\u003e (tomato plant), focusing on their molecular characteristics, tissue-specific expression, and response to abiotic stress. While 58 APs were identified in \u003cem\u003eS. lycopersicum\u003c/em\u003e, only five (AP V, AP W, AP X, AP Y, and AP Z) contained a C-terminal vacuolar sorting domain and the Plant Specific Insert (PSI) domain, a unique domain implicated in vacuolar trafficking and stress responses. These PSI-containing APs were further analyzed through phylogenetics, revealing their evolutionary relationships with homologous APs from other species. Expression profiling showed that most PSI-containing APs are highly expressed in cotyledons and roots, with AP Y being distinctively predominant in reproductive tissues. Under osmotic and saline stress, expression patterns varied, with AP Z exhibiting a broader stress sensitivity. Biochemical analyses confirmed that severe osmotic stress led to increased oxidative stress markers and antioxidant responses, suggestive of activation of the plant-adaptive mechanisms to abiotic stress. 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