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Bamboo is easy to cultivate, grows quickly, and can replace wood or other products after processing. However, the previous layout was mainly based on manual experience. There were issues such as long material transportation distances and frequent interference in transportation paths, resulting in high construction and maintenance costs for production lines. In order to solve the problems, in this work, a new methodology and mathematical formulation was proposed to minimize the total material handling cost and workflow interference. The new method combined a NSGA- II and the A* algorithm in order to minimize the above two objectives. Compared to the Manhattan distance used in traditional algorithms, this method used the A* algorithm to calculate the optimization objective, which can achieve lower material transportation costs and has the ability to handle transportation obstacles and path interference issues. The results showed that after optimization, the average material transportation cost was reduced by 18.1%, with a maximum reduction of 10.5% and a maximum reduction of 97.5% in cross interference points. This indicated that the method proposed in this article was effective in the layout of the original bamboo primary processing production line. [ABSTRACT FROM AUTHOR]