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However, the molecular mechanisms governing their uptake and metabolic integration remain unknown. In this study, we investigated the role of glucosinolate transporters (GTRs) in the active internalization of exogenous GSLs and their subsequent impact on the early development of broccoli (Brassica oleracea L. var. italica) seedlings. Using a multidisciplinary approach combining Xenopus laevis oocyte heterologous expression, confocal microscopy with fluorescent GSL derivatives, and RNA-seq analysis, we demonstrated that broccoli seedlings actively uptake exogenous GSLs through the root system. Accordingly, functional characterization confirmed that BolGTR1.2, BolGTR2.2, BolGTR2.3, and BolGTR3.3 are high-efficiency transporters capable of translocating both aliphatic and indolic GSLs. Together, GSL profiling, time-course RT-qPCR, and Day 7 RNA-seq revealed that GSL internalization triggers a biphasic response: an initial \u201cconsumption phase,\u201d followed by a compensatory upregulation of de novo GSL biosynthetic machinery by Day 7. This metabolic \u201cshortcut\u201d bypasses the energetically costly sulfate reduction pathway, allowing for the reallocation of cellular energy toward structural growth, which was evidenced by the induction of cell wall remodeling genes (XTH, PAE8, GAE) and a measurable increase in seedling biomass. Collectively, our findings support the hypothesis that GSLs are not merely defensive compounds but dynamic metabolic reserves that coordinate resource allocation.<\/jats:p>","DOI":"10.1093\/plphys\/kiag556","type":"journal-article","created":{"date-parts":[[2026,7,29]],"date-time":"2026-07-29T11:18:13Z","timestamp":1785323893000},"source":"Crossref","is-referenced-by-count":0,"title":["Beyond defense: GTR-mediated uptake of glucosinolates drives growth and structural reprogramming in broccoli seedlings"],"prefix":"10.1093","volume":"201","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3461-1534","authenticated-orcid":false,"given":"Lorena","family":"Albaladejo-Marico","sequence":"first","affiliation":[{"name":"Aquaporins Group, Centro de Edafologia y Biologia Aplicada del Segura, CEBAS-CSIC , Campus Universitario de Espinardo 25, Murcia E-30100 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Aplicada del Segura, CEBAS-CSIC , Campus Universitario de Espinardo 25, Murcia E-30100 ,","place":["Spain"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-7659-4316","authenticated-orcid":false,"given":"Josip","family":"Tomas","sequence":"additional","affiliation":[{"name":"GlycoBio&Chemistry Team, ICOA, UMR 7311, Universit\u00e9 D\u2019Orl\u00e9ans et CNRS , BP 6759, Orl\u00e9ans F-45067 ,","place":["France"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1273-0257","authenticated-orcid":false,"given":"Marie","family":"Schuler","sequence":"additional","affiliation":[{"name":"GlycoBio&Chemistry Team, ICOA, UMR 7311, Universit\u00e9 D\u2019Orl\u00e9ans et CNRS , BP 6759, Orl\u00e9ans F-45067 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