Main conclusion <p>Inoculation with <i>Pseudomonas simiae</i> WCS417 improves cucumber growth under Fe deficiency conditions and induces iron-deficiency responses, making it a promising candidate for sustainable biofertilization strategies in dicot plants.</p> Abstract <p>Iron (Fe) deficiency poses a significant agronomic challenge in calcareous soils, particularly affecting dicot plants. Conventional production methods rely heavily on high-yielding varieties and the application of substantial amounts of agrochemicals, leading to considerable environmental concerns. In this context, leveraging the potential of beneficial rhizosphere microorganisms as biofertilizers represents a highly promising and environmentally sound alternative to chemical fertilizers. This study aims to investigate the efficacy of the nonpathogenic strain <i>Pseudomonas simiae</i> WCS417 in eliciting Fe deficiency responses in cucumber plants, along with its impacts on plant growth and Fe chlorosis. Conducted under hydroponic conditions, our experiments reveal compelling outcomes. Root inoculation of cucumber plants with <i>P. simiae</i> significantly enhances plant growth while concurrently mitigating Fe chlorosis symptoms over successive cultivation days. The inoculation with this bacterium induces acidification in the subapical zone of cucumber roots, facilitating Fe solubility in the rhizosphere. Additionally, <i>P. simiae</i> triggers the upregulation of Fe-related genes in inoculated plants, even under Fe sufficiency. In conclusion, <i>P. simiae</i> emerges as a potent enhancer of Fe deficiency responses in cucumber plants. Its ability to promote growth, enhance Fe solubility through rhizosphere acidification, and alleviate Fe chlorosis underscores its potential as an effective biofertilizer for a sustainable Fe nutrition of dicot plants.</p>