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Although iron is an important metal ion for most organisms, the regulatory effect of iron on soil-borne RSSC gene expression remains unknown. Thus, we identified two ferric uptake regulator (Fur) proteins, Fur1 and Fur2, in\n                      <jats:italic toggle=\"yes\">R. pseudosolanacearum<\/jats:italic>\n                      strain OE1-1. All four RSSC phylotype strains had two Fur homologs, a conserved Fur (Fur1) and an alternative Fur (Fur2). We also analyzed the transcriptomes of a\n                      <jats:italic toggle=\"yes\">fur1<\/jats:italic>\n                      deletion mutant,\n                      <jats:italic toggle=\"yes\">fur2<\/jats:italic>\n                      deletion mutant, and\n                      <jats:italic toggle=\"yes\">fur1<\/jats:italic>\n                      \u2044\n                      <jats:italic toggle=\"yes\">fur2<\/jats:italic>\n                      double deletion mutant, revealing that Fur1 and Fur2 cooperatively repress siderophore-related gene expression under Fe\n                      <jats:sup>2+<\/jats:sup>\n                      -rich conditions. Furthermore, extracellular Fe\n                      <jats:sup>3+<\/jats:sup>\n                      -chelating activity was cooperatively controlled by Fur1 and Fur2 under Fe\n                      <jats:sup>2+<\/jats:sup>\n                      -rich conditions. Additionally, nitrate metabolism-related gene expression levels were upregulated only in the\n                      <jats:italic toggle=\"yes\">fur1<\/jats:italic>\n                      \u2044\n                      <jats:italic toggle=\"yes\">fur2<\/jats:italic>\n                      double deletion mutant under Fe\n                      <jats:sup>2+<\/jats:sup>\n                      -rich conditions. This double deletion mutant had a lower growth level than either single deletion mutant. Virulence assays involving tomato plants revealed that single deletions (\n                      <jats:italic toggle=\"yes\">fur1<\/jats:italic>\n                      or\n                      <jats:italic toggle=\"yes\">fur2<\/jats:italic>\n                      ) moderately decreased virulence, whereas the double deletion (\n                      <jats:italic toggle=\"yes\">fur1<\/jats:italic>\n                      and\n                      <jats:italic toggle=\"yes\">fur2<\/jats:italic>\n                      ) resulted in a significant decrease in virulence. Considered together, these results suggest that the two Fur homologs cooperatively regulate gene expression under Fe\n                      <jats:sup>2+<\/jats:sup>\n                      -rich conditions to control strain OE1-1 iron uptake, growth, and virulence.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>IMPORTANCE<\/jats:title>\n                    <jats:p>\n                      The\n                      <jats:italic toggle=\"yes\">Ralstonia solanacearum<\/jats:italic>\n                      species complex (RSSC) comprising soil-borne Gram-negative phytopathogenic bacteria causes bacterial wilt diseases of diverse crop plants. Considering that phylotype I strain OE1-1 enters iron-rich roots from iron-deficient soil during an infection of tomato plants, the mechanisms controlling strain OE1-1 gene expression in response to extracellular iron levels should be clarified. In this study, RSSC was revealed to have two ferric uptake regulator homologs (Fur1 and Fur2). Notably, Fur1 and Fur2 cooperatively repress the expression of genes related to siderophores (Fe\n                      <jats:sup>3+<\/jats:sup>\n                      -chelating compounds) as well as the extracellular Fe\n                      <jats:sup>3+<\/jats:sup>\n                      -chelating activity in the presence of sufficient amounts of extracellular Fe\n                      <jats:sup>2+<\/jats:sup>\n                      . Additionally, Fur1 and Fur2 contribute to the virulence of strain OE1-1 in tomato plants. These findings suggest that RSSC uses two Fur proteins to modulate extracellular Fe\n                      <jats:sup>3+<\/jats:sup>\n                      -chelating activities in response to extracellular iron levels to maintain virulence in crop plants.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1128\/aem.02276-25","type":"journal-article","created":{"date-parts":[[2026,3,2]],"date-time":"2026-03-02T14:00:36Z","timestamp":1772460036000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":1,"title":["Two ferric uptake regulator proteins in\n                    <i>Ralstonia pseudosolanacearum<\/i>\n                    strain OE1-1 function cooperatively in response to the extracellular iron level under ferrous iron-rich 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