Abstract: Seed storability is critical for crop propagation, global grain preservation, and food security. However, knowledge of the molecular biology underlying seed storability remains elusive. Here, we identified the flavonoid 3′ hydroxylase gene (F3′H) as a key gene governing seed storability in soybean through map-based cloning. Functional characterization revealed that GmF3′H not only modulates seed coat pigmentation but, more importantly, also reinforces the physical integrity of both seed coat and hilum, thereby preventing mechanical cracking. Furthermore, conserved F3′H function in soybean and Arabidopsis suggests its broad evolutionary relevance for seed storability. Our findings demonstrate that GmF3′H confers a dual-defense strategy combining structural reinforcement of seed protective layers with antioxidant enhancement for engineering seed longevity and storability. Based on these findings, we developed improved soybean germplasm with enhanced storability by selecting for favorable GmF3′H haplotypes, utilizing linked seed coat color as a visual marker. This study provides new insights for understanding plant seed longevity and offers a valuable avenue for broad-spectrum breeding applications on crop seed storability to sustain seed vigor and grain security.

Key words: map‐based gene cloning, seed coat and hilum, seed storability and longevity, structure robustness