Stoma Detection in Soybean Leaves and Rust Resistance Analysis.

Stomata play a crucial role in plant immune responses, with their morphological characteristics closely linked to disease resistance. Accurate detection and analysis of stomatal phenotypic parameters are essential for soybean disease resistance research and variety breeding. However, traditional stoma detection methods are challenged by complex backgrounds and leaf vein structures in soybean images. To address these issues, we proposed a Soybean Stoma-YOLO (You Only Look Once) model (SS-YOLO) by incorporating large separable kernel attention (LSKA) in the Spatial Pyramid Pooling-Fast (SPPF) module of YOLOv8 and Deformable Large Kernel Attention (DLKA) in the Neck part. These architectural modifications enhanced YOLOV8's ability to extract multi-scale and irregular stomatal features, thus improving detection accuracy. Experimental results showed that SS-YOLO achieved a detection accuracy of 98.7%. SS-YOLO can effectively extract the stomatal features (e.g., length, width, area, and orientation) and calculate related indices (e.g., density, area ratio, variance, and distribution). Across different soybean rust disease stages, the variety Dandou21 (DD21) exhibited less variation in length, width, area, and orientation compared with Fudou9 (FD9) and Huaixian5 (HX5). Furthermore, DD21 demonstrated greater uniformity in stomatal distribution (SEve: 1.02-1.08) and a stable stomatal area ratio (0.06-0.09). The analysis results indicate that DD21 maintained stable stomatal morphology with rust disease resistance. This study demonstrates that SS-YOLO significantly improved stoma detection and provided valuable insights into the relationship between stomatal characteristics and soybean disease resistance, offering a novel approach for breeding and plant disease resistance research.