The design and application of the mushroom picking robot will greatly reduce the labor cost, and it has become one of the research hotspots in the world. Therefore, we independently developed an A. bisporus (a kind of mushroom) picking robot and introduced its functional principle in this paper. At the same time, in order to improve the picking efficiency of the picking robot, a picking path optimization algorithm based on TSP model is proposed. Firstly, based on the TSP model, a picking route model for A. bisporus was established to determine the storage location of each A. bisporus. Then, an improved simulated annealing (I-SA) search algorithm is proposed to find the optimal path sequence. By improving the path initialization module, path generation module, and temperature drop module, the I-SA search algorithm can optimize the picking path in a short time. Finally, in order to improve the stability and reduce the running time of the I-SA search algorithm, a parallel optimization method for global search (“rough exploration”) and local search (“precision exploration”) is proposed. Through simulation experiments, the I-SA search algorithm can search stable and excellent path solution in a relatively short time. Through field experiments on mushroom base, the efficiency of picking A. bisporus can be improved by 14% to 18%, which verifies the effectiveness of the I-SA search algorithm.