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The concentration of dhurrin in sorghum is influenced by both nitrogen status and stage of plant organ development. While nitrogen resupply activates the expression of genes for dhurrin biosynthesis, the molecular mechanisms underlying this regulation remain unclear. In this study, we investigated the transcriptional response of sorghum to nitrogen resupply following growth under nitrogen\u2010limiting conditions. Using a time\u2010course design, we measured hydrogen cyanide potential (HCNp), growth, and nitrate content at 0\u2010, 2\u2010, 6\u2010, 12\u2010, 24\u2010, 36\u2010, 48\u2010, and 60\u2010h after resupply and collected tissue for RNAseq analysis in parallel for analysis of gene expression and construction of gene regulatory networks (GRNs). HCNp (mg\u2009g\n                    <jats:sup>\u22121<\/jats:sup>\n                    DW) increased significantly in leaf and stem tissues following nitrogen resupply, with increases in the leaf partially driven by continued declines in controls under ongoing nitrogen stress. Expression of the dhurrin pathway genes was upregulated in leaves from 24\u2009h after nitrogen resupply, with diel expression patterns observable over the remaining time points. No upregulation was observed in roots or stems, suggesting that developmental context overrides environmental cues. GRN analysis identified candidate transcription factors regulating dhurrin biosynthesis genes, including members of the MYB, bZIP, and GARP\u2010type transcription factor families. Some of these candidate transcription factors may be involved in relieving senescence\u2010associated suppression of dhurrin biosynthesis and link nitrogen signaling to pathway activation. These findings provide new insight into the nitrogen\u2010responsive regulation of dhurrin in sorghum, highlighting candidate regulators for future functional characterization.\n                  <\/jats:p>","DOI":"10.1002\/pld3.70183","type":"journal-article","created":{"date-parts":[[2026,7,28]],"date-time":"2026-07-28T13:13:01Z","timestamp":1785244381000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Nitrogen Status Rewires Transcriptional Regulation of Dhurrin, a Dual\u2010Purpose Defense Metabolite in\n                    <scp>\n                      <i>Sorghum bicolor<\/i>\n                    <\/scp>"],"prefix":"10.1002","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6793-4551","authenticated-orcid":false,"given":"Alicia A.","family":"Quinn","sequence":"first","affiliation":[{"name":"Monash University  Clayton Victoria Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7105-9649","authenticated-orcid":false,"given":"Brian A.","family":"McKinley","sequence":"additional","affiliation":[{"name":"Texas A&amp;M University  College Station Texas USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1919-6982","authenticated-orcid":false,"given":"Cecilia K.","family":"Blomstedt","sequence":"additional","affiliation":[{"name":"Monash University  Clayton Victoria Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2502-2671","authenticated-orcid":false,"given":"John E.","family":"Mullet","sequence":"additional","affiliation":[{"name":"Texas A&amp;M University  College Station Texas USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4756-0411","authenticated-orcid":false,"given":"Roslyn M.","family":"Gleadow","sequence":"additional","affiliation":[{"name":"Monash University  Clayton Victoria Australia"},{"name":"University of Queensland  St Lucia Queensland Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,7,28]]},"reference":[{"key":"e_1_2_12_2_1","doi-asserted-by":"publisher","DOI":"10.1104\/pp.94.3.1219"},{"key":"e_1_2_12_3_1","doi-asserted-by":"publisher","DOI":"10.1111\/tpj.12618"},{"key":"e_1_2_12_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.pbi.2022.102173"},{"key":"e_1_2_12_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.pbi.2020.01.010"},{"key":"e_1_2_12_6_1","doi-asserted-by":"publisher","DOI":"10.1023\/A:1005915507497"},{"key":"e_1_2_12_7_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1438\u20108677.2008.00088.x"},{"key":"e_1_2_12_8_1","doi-asserted-by":"publisher","DOI":"10.1093\/jxb\/eru119"},{"key":"e_1_2_12_9_1","doi-asserted-by":"publisher","DOI":"10.1111\/tpj.13923"},{"key":"e_1_2_12_10_1","doi-asserted-by":"publisher","DOI":"10.1071\/FP17227"},{"key":"e_1_2_12_11_1","doi-asserted-by":"publisher","DOI":"10.3389\/fpls.2021.643192"},{"key":"e_1_2_12_12_1","doi-asserted-by":"publisher","DOI":"10.2135\/cropsci2000.4051295x"},{"key":"e_1_2_12_13_1","doi-asserted-by":"publisher","DOI":"10.1111\/nph.12869"},{"key":"e_1_2_12_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0031-9422(00)90252-8"},{"key":"e_1_2_12_15_1","doi-asserted-by":"publisher","DOI":"10.2134\/agronj1938.00021962003000070004x"},{"key":"e_1_2_12_16_1","doi-asserted-by":"publisher","DOI":"10.2135\/cropsci2012.09.0520"},{"key":"e_1_2_12_17_1","doi-asserted-by":"publisher","DOI":"10.1104\/pp.000687"},{"key":"e_1_2_12_18_1","doi-asserted-by":"publisher","DOI":"10.1038\/ncomms12570"},{"key":"e_1_2_12_19_1","doi-asserted-by":"publisher","DOI":"10.1080\/00103627509366547"},{"key":"e_1_2_12_20_1","doi-asserted-by":"publisher","DOI":"10.1111\/jipb.12899"},{"key":"e_1_2_12_21_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkaf018"},{"key":"e_1_2_12_22_1","doi-asserted-by":"publisher","DOI":"10.1104\/pp.112.198705"},{"key":"e_1_2_12_23_1","doi-asserted-by":"publisher","DOI":"10.3390\/agronomy14092073"},{"key":"e_1_2_12_24_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gky1081"},{"key":"e_1_2_12_25_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.phytochem.2022.113427"},{"key":"e_1_2_12_26_1","doi-asserted-by":"publisher","DOI":"10.1038\/srep37079"},{"key":"e_1_2_12_27_1","doi-asserted-by":"publisher","DOI":"10.1016\/s1673\u20108527(09)60003\u20105"},{"key":"e_1_2_12_28_1","doi-asserted-by":"publisher","DOI":"10.3390\/ijms21041445"},{"key":"e_1_2_12_29_1","doi-asserted-by":"publisher","DOI":"10.1071\/fp24343"},{"key":"e_1_2_12_30_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev\u2010arplant\u2010050718\u2010095910"},{"key":"e_1_2_12_31_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365\u2010313X.2006.02715.x"},{"key":"e_1_2_12_32_1","volume-title":"Regulation of Dhurrin Synthesis in Forage Sorghum. 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