Early cadmium responses in developing oat caryopses indicate an unexpected regulatory network linked to low grain cadmium accumulation

Heavy-metal accumulation in cereal grains is becoming critical for European food safety, regulation and plant breeding. In the EU, Cd maximum levels in certain foodstuffs have been revised, including lowering or establishing limits for relevant food categories, while new maximum levels for nickel (Ni) have recently been introduced for several foodstuffs, including cereal categories, with limits for oats and selected cereals applying from 2026. Together, these developments create an urgent need to identify genetic and physiological mechanisms that reduce Cd and Ni accumulation in cereal grains while maintaining crop quality and productivity. Against this regulatory and food-safety background, our broader RNA-seq experiment investigates early transcriptional responses to Cd and Ni in oat F2 segregants contrasting for metal accumulation. The full dataset includes low- and high-accumulating segregants, roots and developing caryopses sampled at 3 h and 7 h after treatment. In the present pilot analysis, we focus on the Cd response in developing caryopses of the low-Cd accumulating segregant AS131 to identify candidate processes associated with reduced grain Cd accumulation. The strongest transcriptional responses were not dominated by canonical Cd-detoxification pathways. At 3 h after Cd exposure, differentially expressed transcripts were mainly associated with cell-wall functions, endosperm transfer-cell-specific PR60 proteins, DUF239-containing proteins and cysteine proteinase inhibitors, whereas several dehydration-, pathogen-, defence-, cell-wall-loosening- and ROS- related genes were repressed. By 7 h, the response suggested a shift towards homeostatic acclimation, with induction of TIP2 aquaporins, thiamine thiazole synthases, EF-Tu proteins, coatomer-related genes and carbohydrate metabolism-associated genes, together with repression of LEA/SMP/dehydrin genes, FRO7-like genes, EF-hand calcium-binding proteins and stress-regulatory transcription factors. Pathway-level analyses were broadly consistent with these transcript-level patterns, highlighting structural, nucleosome-associated, translation-related, metabolic and developmental processes. Several Cd-responsive transcripts were also associated with broader abiotic-stress responses, suggesting recruitment of shared stress-regulatory modules rather than Cd-specific detoxification pathways alone. Overall, these results support a working hypothesis in which low Cd accumulation in developing oat grain may involve regulation of solute-transfer interfaces, cellular protection, intracellular homeostasis, trafficking pathways and caryopsis developmental programmes. These findings provide candidate processes for future comparison with high-Cd accumulating segregants, root tissues and Ni responses in the broader dataset.