Advanced phenotyping features utilizing deep learning techniques for automated analysis of stomatal guard cell orientation.

Stomata are vital for controlling gas exchange and water vapor release, which significantly affect photosynthesis and transpiration. Characterizing stomatal traits such as size, density, and distribution is essential for adaptation to the environment. While microscopy is widely used for this purpose, manual analysis is labor-intensive and time-consuming that limit large scale studies. To overcome this, we introduce an automated, high-throughput method that leverages YOLOv8, an advanced deep learning model, for more accurate and efficient stomatal trait measurement. Our approach provides a comprehensive analysis of stomatal morphology by examining both stomatal pores and guard cells. A key finding is the introduction of stomatal angles as a novel phenotyping trait, which can offer deeper insights into stomatal function. We developed a model using a carefully annotated dataset that accurately segments and analyzes stomatal guard cells from high-resolution images. Additionally, our study introduces a new opening ratio metric, calculated from the areas of the guard cells and the stomatal pore, providing a valuable morphological descriptor for future physiological research. This scalable system significantly enhances the precision and efficiency of large-scale plant phenotyping, offering a new tool to advance research in plant physiology.