{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,9]],"date-time":"2026-05-09T00:09:45Z","timestamp":1778285385110,"version":"3.51.4"},"reference-count":18,"publisher":"Public Library of Science (PLoS)","issue":"5","license":[{"start":{"date-parts":[[2026,5,8]],"date-time":"2026-05-08T00:00:00Z","timestamp":1778198400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:p>Conventional selectable marker genes (SMGs) in plant transformation, such as those conferring antibiotic resistance, raise biosafety concerns due to their permanent genomic integration. Alternative systems, such as the phosphite (Phi)\/phosphite dehydrogenase (PtxD) system, have been developed to address these concerns. This system works by enabling transgenic plants to convert non-metabolizable, toxic Phi into usable orthophosphate (Pi), thereby providing both a detoxification mechanism and a positive growth advantage for selection. Building on this principle of detoxification-based positive selection, we have developed a novel system utilizing the microbial biuret hydrolase (BH) gene and its substrate, biuret (BU). Biuret is a phytotoxic by-product found in urea fertilizers. We introduced two genetic constructs into tobacco via Agrobacterium-mediated transformation: one expressing only the BH enzyme and another expressing a dual-function BH\u2013AH fusion protein (BA2H, where AH is allophanate hydrolase). On nitrate-containing media, selection with 1 mM BU achieved a transformation efficiency of nearly 80% and shortened regeneration time by approximately 20 days compared to conventional Kanamycin selection (66.7% efficiency). The resulting transgenic lines demonstrated strong tolerance to BU during germination and seedling growth, even under nitrogen-deficient conditions. BU also acted as a selective herbicide, reducing weed biomass by ~70% in co-culture and field-simulation experiments while transgenic plants thrived. The BH\/BU system provides an efficient, safe, and agriculturally relevant selection platform that avoids antibiotic resistance markers and leverages BU\u2019s dual role as a nitrogen source for crops and a growth inhibitor for weeds. This approach offers a sustainable alternative to conventional selection markers in plant biotechnology.<\/jats:p>","DOI":"10.1371\/journal.pone.0347957","type":"journal-article","created":{"date-parts":[[2026,5,8]],"date-time":"2026-05-08T23:11:03Z","timestamp":1778281863000},"page":"e0347957","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":0,"title":["A novel positive selection system for plant transformation based on microbial biuret hydrolase and 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