AbstractPine wilt disease (PWD) is currently one of the main causes of large-scale forest destruction. To control the spread of PWD, it is essential to detect affected pine trees quickly. This study investigated the feasibility of using the object-oriented multi-scale segmentation algorithm to identify trees discolored by PWD. We used an unmanned aerial vehicle (UAV) platform equipped with an RGB digital camera to obtain high spatial resolution images, and multi-scale segmentation was applied to delineate the tree crown, coupling the use of object-oriented classification to classify trees discolored by PWD. Then, the optimal segmentation scale was implemented using the estimation of scale parameter (ESP2) plug-in. The feature space of the segmentation results was optimized, and appropriate features were selected for classification. The results showed that the optimal scale, shape, and compactness values of the tree crown segmentation algorithm were 56, 0.5, and 0.8, respectively. The producer’s accuracy (PA), user’s accuracy (UA), and F1 score were 0.722, 0.605, and 0.658, respectively. There were no significant classification errors in the final classification results, and the low accuracy was attributed to the low number of objects count caused by incorrect segmentation. The multi-scale segmentation and object-oriented classification method could accurately identify trees discolored by PWD with a straightforward and rapid processing. This study provides a technical method for monitoring the occurrence of PWD and identifying the discolored trees of disease using UAV-based high-resolution images.