Asynchronous origins of Yellow-Stripe transporters and Nicotianamine Synthase underlie the evolution of plant Strategy-II iron uptake

Plants acquire iron using two canonical mechanisms: reduction-based uptake (Strategy I) and chelation-based uptake (Strategy II). Although Strategy II is characteristic of grasses, genes associated with chelated-metal transport occur broadly across plant lineages, obscuring how this pathway evolved. Here, we reconstruct the evolutionary history of Yellow Stripe-Like (YSL) transporters and Nicotianamine Synthase (NAS) across Archaeplastida and their closest non-plant homologs. Phylogenomic, distributional, and structural analyses reveal pronounced temporal uncoupling between these gene families. YSL most likely originated early in Viridiplantae and represents the deepest evolutionary module of the chelated-metal transport system. In contrast, NAS appeared much later through independent horizontal gene transfer events from fungi into euphyllophytes and specific moss lineages. This timing indicates that YSL-mediated transport initially functioned independently of nicotianamine, implying ancestral use of alternative siderophores. A pronounced expansion of YSL genes in Poaceae coincides with the emergence of canonical Strategy II, suggesting that this pathway arose by exaptation of pre-existing chelated-metal transport mechanisms.