Stem diameter is a critical phenotypic parameter for maize, integral to yield prediction and lodging resistance assessment. Traditionally, the quantification of this parameter through manual measurement has been the norm, notwithstanding its tedious and laborious nature. To address these challenges, this study introduces a non-invasive field-based system utilizing depth information from RGB-D cameras to measure maize stem diameter. This technology offers a practical solution for conducting rapid and non-destructive phenotyping. Firstly, RGB images, depth images, and 3D point clouds of maize stems were captured using an RGB-D camera, and precise alignment between the RGB and depth images was achieved. Subsequently, the contours of maize stems were delineated using 2D image processing techniques, followed by the extraction of the stem's skeletal structure employing a thinning-based skeletonization algorithm. Furthermore, within the areas of interest on the maize stems, horizontal lines were constructed using points on the skeletal structure, resulting in 2D pixel coordinates at the intersections of these horizontal lines with the maize stem contours. Subsequently, a back-projection transformation from 2D pixel coordinates to 3D world coordinates was achieved by combining the depth data with the camera's intrinsic parameters. The 3D world coordinates were then precisely mapped onto the 3D point cloud using rigid transformation techniques. Finally, the maize stem diameter was sensed and determined by calculating the Euclidean distance between pairs of 3D world coordinate points. The method demonstrated a Mean Absolute Percentage Error ( MAPE ) of 3.01%, a Mean Absolute Error ( MAE ) of 0.75 mm, a Root Mean Square Error ( RMSE ) of 1.07 mm, and a coefficient of determination ( R ²) of 0.96, ensuring accurate measurement of maize stem diameter. This research not only provides a new method of precise and efficient crop phenotypic analysis but also offers theoretical knowledge for the advancement of precision agriculture., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Zhou, Cui, Wu, Gao, Tang, Jiang, Wu, Zhang and Hou.)