Enhancing Iron Nutrition in Citrus: Synergistic Roles of Proline-2′-deoxymugineic Acid in Root Physiology and Microbiome

Iron (Fe) deficiency severely impairs plant growth and development in calcareous soils. Proline-2′-deoxymugineic acid (PDMA), a phytosiderophore analog that enhances Fe availability, alleviates Fe deficiency in field and vegetable crops but remains untested in perennial woody crops. Herein, we conducted pot and field trials on citrus, integrating physiological assays, RNA sequencing, 16S rRNA profiling, and metagenomics to evaluate PDMA/PDMA–Fe(III) effects on Fe nutrition, yield, root gene expression, and rhizosphere microbial dynamics. Results showed that PDMA/PDMA–Fe(III) significantly improved citrus Fe nutrition-outperforming traditional EDTA–Fe(III)– by increasing rhizosphere Fe availability, thereby increasing yield and downregulating Fe uptake- and stress response-related genes,with PDMA–Fe(III) had stronger suppression. PDMA–Fe(III) minimally disrupted the rhizosphere microbiome, while PDMA recruited plant growth-promoting rhizobacteria (e.g., Pseudomonas, Nitrospira ); both treatments enriched microbial carbon fixation pathways. Collectively, PDMA/PDMA–Fe(III) represent eco-efficient Fe fertilizers for citrus orchards, providing sustainable remediation of Fe deficiency in calcareous soils.