Deep aerenchyma: a transformer-based pipeline for scalable phenotyping of rice root aerenchyma lacunae across environments

BACKGROUND: Quantification of rice root anatomical traits such as cortical aerenchyma lacunae is key to understanding rice adaptation to diverse water regimes and to support climate-smart breeding. Aerenchyma lacunae contributes to rice internal gas transport and influences methane emissions from flooded systems and can also limit rice water conductivity. It could be an interesting anatomical trait for breeding, however, large-scale anatomical phenotyping remains limited because manual analysis of root cross-sections is labor-intensive, subjective, and difficult to scale across heterogeneous imaging conditions. Existing pipelines often require parameter tuning and do not generalize well across environments. RESULTS: We developed a deep learning pipeline based on a vision transformer architecture to automatically segment rice root cross-sections and quantify cortical aerenchyma lacunae. The model was trained on 1,760 annotated images collected across multiple countries, growth stages, cultivation systems, and experimental contexts, using a collaboratively defined annotation protocol. The final model achieved high segmentation accuracy, with mean intersection over union values exceeding 0.92 for cortical tissues and lacunae. Quantification of the lacuna-to-cortex ratio showed strong agreement with manual annotations, with a coefficient of determination of 0.98 on an independent test set. An independent expert review indicated that model predictions were at least as consistent as manual annotations and reduced large annotation inconsistencies. The pipeline is released as open-source software and includes an interactive online demonstrator, and is accompanied by an online test dataset to support testing and reproducibility. Application across six experimental use cases revealed reproducible differences in aerenchyma lacunae across genotypes, water regimes, environments, and developmental stages. CONCLUSIONS: This work provides a robust, scalable, and transferable tool for automated root anatomical phenotyping under heterogeneous experimental conditions. Transformer-based segmentation enables consistent and high-throughput quantification of lacunae, facilitating integration of these anatomical traits into breeding, physiological studies, and climate-smart crop improvement programs.