Aiming at the problem of wildlife-vehicle collisions and traffic efficiency of road sections influenced by wildlife crossings on the ecological roads, the multi-objective optimization of intersection signal control and traffic control models is studied. To mitigate the risk of wildlife-vehicle collisions, virtualizing wildlife crossing behaviors as two-phase signal lights on road segments is proposed, thereby a traffic control model for animal crossing ecological roads is constructed. A signal timing optimization model that considers parameters such as maximum queue length at intersections, downstream road smoothness and density is established by utilizing the vehicle speed and position information within the V2X ecological road traffic framework. This model aims to enhance construction vehicle passage efficiency at ecological road intersections and improve the smoothness of downstream segments. The signal timing optimization strategy is solved using a genetic algorithm. To validate the effectiveness of the proposed model and algorithm, simulation experiments are conducted and results are compared using a practical case study. The results demonstrate that the optimization objective value (S) increases by 20%, the maximum queue length at intersections decreases by 33.3%, and the vehicle density on downstream segments reduces by 15.4%. These results confirm the efficacy of the proposed model and algorithm in achieving intersection signal optimization control for ecological roads under V2X, reducing the maximum vehicle queue length at intersections, and decreasing vehicle travel time.