{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,12]],"date-time":"2026-07-12T04:59:33Z","timestamp":1783832373695,"version":"3.55.0"},"reference-count":50,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2020,8,25]],"date-time":"2020-08-25T00:00:00Z","timestamp":1598313600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["31470347 and 31770269"],"award-info":[{"award-number":["31470347 and 31770269"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Plants"],"abstract":"<jats:p>Iron (Fe) homeostasis is essential for plant growth and development, and it is strictly regulated by a group of transcriptional factors. Iron-related transcription factor 3 (OsIRO3) was previously identified as a negative regulator for Fe deficiency response in rice. However, the molecular mechanisms by which OsIRO3 regulate Fe homeostasis is unclear. Here, we report that OsIRO3 is essential for responding to Fe deficiency and maintaining Fe homeostasis in rice. OsIRO3 is expressed in the roots, leaves, and base nodes, with a higher level in leaf blades at the vegetative growth stage. Knockout of OsIRO3 resulted in a hypersensitivity to Fe deficiency, with severe necrosis on young leaves and defective root development. The iro3 mutants accumulated higher levels of Fe in the shoot under Fe-deficient conditions, associated with upregulating the expression of OsNAS3, which lead to increased accumulation of nicotianamine (NA) in the roots. Further analysis indicated that OsIRO3 can directly bind to the E-box in the promoter of OsNAS3. Moreover, the expression of typical Fe-related genes was significantly up-regulated in iro3 mutants under Fe-sufficient conditions. Thus, we conclude that OsIRO3 plays a key role in responding to Fe deficiency and regulates NA levels by directly, negatively regulating the OsNAS3 expression.<\/jats:p>","DOI":"10.3390\/plants9091095","type":"journal-article","created":{"date-parts":[[2020,8,26]],"date-time":"2020-08-26T09:05:37Z","timestamp":1598432737000},"page":"1095","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":35,"title":["OsIRO3 Plays an Essential Role in Iron Deficiency Responses and Regulates Iron Homeostasis in Rice"],"prefix":"10.3390","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1206-4119","authenticated-orcid":false,"given":"Wujian","family":"Wang","sequence":"first","affiliation":[{"name":"College of Life Sciences, Nanjing Agricultural University, Nanjing 210095, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jun","family":"Ye","sequence":"additional","affiliation":[{"name":"College of Life Sciences, Nanjing Agricultural University, Nanjing 210095, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanran","family":"Ma","sequence":"additional","affiliation":[{"name":"College of Life Sciences, Nanjing Agricultural University, Nanjing 210095, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ting","family":"Wang","sequence":"additional","affiliation":[{"name":"College of Life Sciences, Nanjing Agricultural University, Nanjing 210095, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huixia","family":"Shou","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Plant Physiology and Biochemistry, College of Life Sciences, Zhejiang University, Hangzhou 310058, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2711-780X","authenticated-orcid":false,"given":"Luqing","family":"Zheng","sequence":"additional","affiliation":[{"name":"College of Life Sciences, Nanjing Agricultural University, Nanjing 210095, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,8,25]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"333","DOI":"10.1038\/nchembio.166","article-title":"Facing the challenges of Cu, Fe and Zn homeostasis in plants","volume":"5","author":"Palmer","year":"2009","journal-title":"Nat. Chem. Biol."},{"key":"ref_2","unstructured":"WHO\/UNICEF\/UNU (2001). Iron Deficiency Anaemia: Assessment, Prevention and Control: A Guide for Programme Managers, WHO\/UNICEF\/UNU. Document WHO\/NHD\/01.03."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1146\/annurev-arplant-042811-105522","article-title":"Iron uptake, translocation, and regulation in higher plants","volume":"63","author":"Kobayashi","year":"2012","journal-title":"Annu. Rev. Plant Biol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"261","DOI":"10.1007\/BF00008069","article-title":"Strategies of plants for acquisition of iron","volume":"165","author":"Marschner","year":"1994","journal-title":"Plant Soil"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"5624","DOI":"10.1073\/pnas.93.11.5624","article-title":"A novel iron-regulated metal transporter from plants identified by functional expression in yeast","volume":"93","author":"Eide","year":"1996","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"694","DOI":"10.1038\/17800","article-title":"A ferric-chelate reductase for iron uptake from soils","volume":"397","author":"Robinson","year":"1999","journal-title":"Nature"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1072","DOI":"10.1111\/j.1469-8137.2009.02908.x","article-title":"Dissecting iron deficiency-induced proton extrusion in Arabidopsis roots","volume":"183","author":"Simonetta","year":"2009","journal-title":"New Phytol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"346","DOI":"10.1038\/35053080","article-title":"Maize yellow stripe1 encodes a membrane protein directly involved in Fe(III) uptake","volume":"409","author":"Curie","year":"2001","journal-title":"Nature"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"563","DOI":"10.1111\/j.1365-313X.2006.02714.x","article-title":"A specific transporter for iron(III)-phytosiderophore in barley roots","volume":"46","author":"Murata","year":"2006","journal-title":"Plant J."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"3470","DOI":"10.1074\/jbc.M806042200","article-title":"Rice OsYSL15 is an iron-regulated iron(III)-deoxymugineic acid transporter expressed in the roots and is essential for iron uptake in early growth of the seedlings","volume":"284","author":"Inoue","year":"2009","journal-title":"J. Biol. Chem."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"5446","DOI":"10.1074\/jbc.M110.180026","article-title":"Phytosiderophore efflux transporters are crucial for iron acquisition in graminaceous plants","volume":"286","author":"Nozoye","year":"2011","journal-title":"J. Biol. Chem."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"3400","DOI":"10.1105\/tpc.104.024315","article-title":"The essential basic Helix-Loop-Helix protein FIT1 is required for the iron deficiency response","volume":"16","author":"Colangelo","year":"2004","journal-title":"Plant Cell"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"385","DOI":"10.1038\/cr.2008.26","article-title":"FIT interacts with AtbHLH38 and AtbHLH39 in regulating iron uptake gene expression for iron homeostasis in Arabidopsis","volume":"18","author":"Yuan","year":"2008","journal-title":"Cell Res."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"790","DOI":"10.1104\/pp.111.190983","article-title":"Co-Overexpression FIT with AtbHLH38 or AtbHLH39 in Arabidopsis-Enhanced cadmium tolerance via increased cadmium sequestration in roots and improved iron homeostasis of shoots","volume":"158","author":"Wu","year":"2012","journal-title":"Plant Physiol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1093\/mp\/ssr065","article-title":"Fitting into the harsh reality: Regulation of iron-deficiency responses in dicotyledonous plants","volume":"5","author":"Ivanov","year":"2012","journal-title":"Mol. Plant"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"2219","DOI":"10.1105\/tpc.110.074096","article-title":"The bHLH transcription factor POPEYE regulates response to iron deficiency in Arabidopsis roots","volume":"22","author":"Long","year":"2010","journal-title":"Plant Cell"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"273","DOI":"10.1104\/pp.114.250837","article-title":"Iron-Binding E3 ligase mediates iron response in plants by targeting basic Helix-Loop-Helix transcription factors","volume":"167","author":"Selote","year":"2014","journal-title":"Plant Physiol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"787","DOI":"10.1105\/tpc.114.132704","article-title":"The bHLH transcription factor bHLH104 interacts with IAA-LEUCINE RESISTANT3 and modulates iron homeostasis in Arabidopsis","volume":"27","author":"Zhang","year":"2015","journal-title":"Plant Cell"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2478","DOI":"10.1104\/pp.15.01827","article-title":"Two bHLH transcription factors, bHLH34 and bHLH104, regulate iron homeostasis in Arabidopsis thaliana","volume":"170","author":"Li","year":"2016","journal-title":"Plant Physiol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1743","DOI":"10.1093\/jxb\/erx043","article-title":"bHLH transcription factor bHLH115 regulates iron homeostasis in Arabidopsis thaliana","volume":"68","author":"Liang","year":"2017","journal-title":"J. Exp. Bot."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1007\/s10265-018-1068-z","article-title":"The basic helix-loop-helix transcription factor, bHLH11 functions in the iron-uptake system in Arabidopsis thaliana","volume":"132","author":"Tanabe","year":"2019","journal-title":"J. Plant Res."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"508","DOI":"10.1105\/tpc.19.00541","article-title":"The transcription factor bHLH121 interacts with bHLH105 (ILR3) and its closest homologs to regulate iron homeostasis in Arabidopsis","volume":"32","author":"Gao","year":"2019","journal-title":"Plant Cell"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"24933","DOI":"10.1073\/pnas.1916892116","article-title":"The iron deficiency response in Arabidopsis thaliana requires the phosphorylated transcription factor URI","volume":"116","author":"Kim","year":"2019","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"634","DOI":"10.1016\/j.molp.2020.01.006","article-title":"bHLH121 functions as a direct link that facilitates the activation of FIT by bHLH IVc transcription factors for maintaining Fe homeostasis in Arabidopsis","volume":"13","author":"Lei","year":"2020","journal-title":"Mol. Plant"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"2867","DOI":"10.1093\/jxb\/erl054","article-title":"Isolation and characterization of IRO2, a novel iron-regulated bHLH transcription factor in graminaceous plants","volume":"57","author":"Ogo","year":"2006","journal-title":"J. Exp. Bot."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"366","DOI":"10.1111\/j.1365-313X.2007.03149.x","article-title":"The rice bHLH protein OsIRO2 is an essential regulator of the genes involved in Fe uptake under Fe-deficient conditions","volume":"51","author":"Ogo","year":"2007","journal-title":"Plant J."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1247","DOI":"10.1111\/nph.16232","article-title":"A transcription factor OsbHLH156 regulates Strategy II iron acquisition through localising IRO2 to the nucleus in rice","volume":"225","author":"Wang","year":"2020","journal-title":"New Phytol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"668","DOI":"10.1111\/jipb.12933","article-title":"Oryza sativa FER-LIKE FE DEFICIENCY-INDUCED TRANSCRIPTION FACTOR (OsFIT\/OsbHLH156) interacts with OsIRO2 to regulate iron homeostasis","volume":"62","author":"Liang","year":"2020","journal-title":"J. Integr. Plant Biol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"166","DOI":"10.1186\/1471-2229-10-166","article-title":"Identification of a novel iron regulated basic helix-loop-helix protein involved in Fe homeostasis in Oryza sativa","volume":"10","author":"Zheng","year":"2010","journal-title":"BMC Plant Biol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"543","DOI":"10.1104\/pp.17.00794","article-title":"Positive regulator of iron homeostasis1, OsPRI1, facilitates iron homeostasis","volume":"175","author":"Zhang","year":"2017","journal-title":"Plant Physiol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"471","DOI":"10.1007\/s11103-019-00917-8","article-title":"OsbHLH058 and OsbHLH059 transcription factors positively regulate iron deficiency responses in rice","volume":"101","author":"Kobayashi","year":"2019","journal-title":"Plant Mol. Biol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"261","DOI":"10.1111\/pce.13655","article-title":"Oryza sativa POSITIVE REGULATOR of IRON DEFICIENCY RESPONSE 2 (OsPRI2) and OsPRI3 are involved in the maintenance of Fe homeostasis","volume":"43","author":"Zhang","year":"2019","journal-title":"Plant Cell Environ."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"224","DOI":"10.1111\/j.1365-3040.2012.02569.x","article-title":"Identification of OsbHLH133 as a regulator of iron distribution between roots and shoots in Oryza sativa","volume":"36","author":"Wang","year":"2013","journal-title":"Plant Cell Environ."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1749","DOI":"10.1105\/tpc.013839","article-title":"The arabidopsis Basic\/Helix-Loop-Helix transcription factor family","volume":"15","year":"2003","journal-title":"Plant Cell"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"6","DOI":"10.3389\/fpls.2019.00006","article-title":"The transcriptional control of iron homeostasis in plants: A tale of bHLH transcription factors?","volume":"10","author":"Gao","year":"2019","journal-title":"Front. Plant Sci."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"909","DOI":"10.1093\/jxb\/erx465","article-title":"Trace metal metabolism in plants","volume":"69","author":"Andresen","year":"2018","journal-title":"J. Exp. Bot."},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Zhang, X., Zhang, D., Sun, W., and Wang, T. (2019). The adaptive mechanism of plants to iron deficiency via iron uptake, transport, and homeostasis. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20102424"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"366","DOI":"10.1046\/j.1365-313X.2003.01878.x","article-title":"Three rice nicotianamine synthase genes, OsNAS1, OsNAS2, and OsNAS3 are expressed in cells involved in long-distance transport of iron and differentially regulated by iron","volume":"36","author":"Inoue","year":"2003","journal-title":"Plant J."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"269","DOI":"10.1104\/pp.108.1.269","article-title":"Subcellular localization and characterization of excessive iron in the nicotianamine-less tomato mutant chloronerva","volume":"108","author":"Becker","year":"1995","journal-title":"Plant Physiol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"355","DOI":"10.1146\/annurev-phyto-080516-035537","article-title":"Iron and Immunity","volume":"55","author":"Verbon","year":"2017","journal-title":"Annu. Rev. Phytopathol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1007\/s11103-007-9262-8","article-title":"Identification and localisation of the rice nicotianamine aminotransferase gene OsNAAT1 expression suggests the site of phytosiderophore synthesis in rice","volume":"66","author":"Inoue","year":"2008","journal-title":"Plant Mol. Biol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1433","DOI":"10.1111\/nph.15753","article-title":"Transcriptional integration of the responses to iron availability in Arabidopsis by the bHLH factor ILR3","volume":"223","author":"Tissot","year":"2019","journal-title":"New Phytol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1975","DOI":"10.1093\/mp\/sst119","article-title":"RNA-Guided genome editing in plants using a CRISPR\u2013cas9 system","volume":"6","author":"Xie","year":"2013","journal-title":"Mol. Plant."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1046\/j.1365-313X.2003.01860.x","article-title":"Distribution and characterization of over 1000 T-DNA tags in rice genome","volume":"36","author":"Chen","year":"2003","journal-title":"Plant J."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"1308","DOI":"10.1094\/PDIS-01-11-0026","article-title":"Comparison of water displacement and WinRHIZO software for plant root parameter assessment","volume":"95","author":"Pang","year":"2011","journal-title":"Plant Dis."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"1586","DOI":"10.1105\/tpc.114.123208","article-title":"SPX4 negatively regulates phosphate signaling and homeostasis through its interaction with PHR2 in rice","volume":"26","author":"Lv","year":"2014","journal-title":"Plant Cell"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1186\/s13007-020-0556-z","article-title":"A native chromatin immunoprecipitation (ChIP) protocol for studying histone modifications in strawberry fruits","volume":"16","author":"Huang","year":"2020","journal-title":"Plant Methods"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"788","DOI":"10.1007\/s10535-015-0554-6","article-title":"Sufficient sulfur supply promotes seedling growth, alleviates oxidation stress, and regulates iron uptake and translocation in rice","volume":"59","author":"Wu","year":"2015","journal-title":"Biol. Plantarum."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"2267","DOI":"10.1105\/tpc.18.00051","article-title":"Alternative splicing plays a critical role in maintaining mineral nutrient homeostasis in rice (Oryza sativa)","volume":"30","author":"Dong","year":"2018","journal-title":"Plant Cell"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"1187","DOI":"10.1046\/j.1365-313X.1997.11061187.x","article-title":"Subcellular localization of H2O2 in plants. H2O2 accumulation in papillae and hypersensitive response during the barley-powdery mildew interaction","volume":"11","author":"Zhang","year":"1997","journal-title":"Plant J."}],"container-title":["Plants"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2223-7747\/9\/9\/1095\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T10:06:35Z","timestamp":1760177195000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2223-7747\/9\/9\/1095"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,8,25]]},"references-count":50,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2020,9]]}},"alternative-id":["plants9091095"],"URL":"https:\/\/doi.org\/10.3390\/plants9091095","relation":{},"ISSN":["2223-7747"],"issn-type":[{"value":"2223-7747","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,8,25]]}}}