Combining RGB imaging with a two-stage deep learning method to reveal genetic variation of wheat sprouting traits

= 0.914 and RMSE = 2.448 compared to manual measurements, and required less than 0.2 s per image for analysis. By employing this two-stage algorithm for processing and analysis, varieties (e.g., Gao8901 and ShiYou20) that consistently exhibited high emergence rates and uniformity under multiple nitrogen treatments were identified. Furthermore, genome-wide association study identified the major loci qEmergence rate-3A and qUniformity-6B governing seedling emergence rate and uniformity, which likely enhance wheat seedling traits by modulating energy supply or related signaling molecules. The emergence-rate and uniformity data generated by the two-stage algorithm significantly accelerated the discovery of relevant genes and enabled the identification of wheat varieties with high emergence rate and uniformity, providing valuable insights and practical references for high-quality breeding and gene mining.