The study optimized three wheat transformation methods—immature embryo, callus, and in planta injection—by systematically adjusting Agrobacterium strain, bacterial density, acetosyringone concentration, and incubation conditions, achieving transformation efficiencies up to 66.84%. Using these protocols, CRISPR/Cas9 knockout of the negative regulator TaARE1-D produced mutants with increased grain number, spike length, grain size, and a stay‑green phenotype, demonstrating the platform’s potential to accelerate yield and stress‑tolerance improvements in wheat.

The study used a yeast two-hybrid screen to identify 52 wheat proteins that interact with the inositol pyrophosphate kinase TaVIH2-3B, highlighting the fasciclin‑like arabinogalactan protein TaFLA7 as a key partner involved in cell‑wall functions. Pulldown assays and reporter fusion analyses confirmed the interaction and plasma‑membrane localization of TaFLA7, which is modulated by TaVIH2‑3B activity and shows drought‑responsive and grain‑development expression in wheat.

The study generated wheat (Triticum aestivum) mutants with targeted deletions in the SAL gene family (TaSAL1 and TaSAL2) to assess the impact of chloroplast-to-nucleus retrograde signaling on field performance. Across 15 diverse Australian field trials, TaSAL2 deletions conferred 4–8% higher yields and improved water productivity by maintaining photosynthetic efficiency and dynamic stomatal control under drought, whereas TaSAL1 deletions reduced yields. These results demonstrate that locus‑specific retrograde signaling modifications can simultaneously enhance yield and stress resilience in a major crop.