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Aiming at the problems of difficulty in examining phenotypic traits and time-consuming and laborious manual measurements during shiitake mushroom breeding, a phenotypic measurement method for shiitake mushroom sticks applicable to the deployment of the Jetson Orin Nano platform was proposed. Image acquisition of shiitake mushroom sticks from three different data sources was performed by using cell phones and industrial cameras, and the shiitake mushroom colony dataset was labeled and enhanced;the segmentation effects of five segmentation models, namely, Bisenet, Stdcseg, U-net, Deeplabv3p, and PP-liteseg, were compared on the Test-A, Test-B, and Test-C test sets, and the results showed that PP-liteseg was the most suitable segmentation model for the test set of shiitake mushroom. The results showed that PP-liteseg was more pervasive than other networks, the mIoU of PP-liteseg segmentation models on the three test sets was more than 97.53%, the mPA was higher than 99.49%, and the inference of a single image took 660 ms. To further balance the accuracy and real-time performance of the model, the PP-liteseg model was compressed by quantized distillation and deployed on the Jetson Orin Nano platform, and the mIoU and mPA of the compressed model on the Test-B test set were 97.50% and 99.51%, respectively, and the inference of a single image took 43.63 ms, which was nearly 64% shorter than that of the pre-compression model;the images of shiitake mushroom sticks were segmented by using the PP-liteseg, and the phenotypes of colonies were extracted, and then the mycelial growth length was obtained based on the radial length and axial width of the colony, and the average absolute percentage error, root mean square error and coefficient of determination of the mycelial growth length were 1.874%, 0.148 cm and 0.918, respectively, compared with the manually measured values;the mycelial growth length of six strains were measured by the method in four consecutive days, and the results showed that the mycelial growth rate of six strains, both in a single day or during the whole culture period, with the greatest difference between strains 49 and 168. The study demonstrated that the method was suitable for the phenotypic measurement of shiitake mushroom mycelium and can be run on the Jetson Orin Nano platform with good accuracy and real-time performance.