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Because of the size and complexity of large space truss structures and multiple space robots environment, on-orbit assembly requires careful planning. An ant colony algorithm-based approach is applied to generate optimal assembly sequences of large space truss structures. The inherent hierarchical nature of large space truss structures allows assembly sequences to be considered from two levels: strut level and structural volume element (SVE) level. At the strut level, assembly sequences of the struts of each structural volume element are represented by the connectivity matrix. Furthermore, at the SVE level, the directed graph representation is used because it can guarantee the feasibility conditions of assembly tasks and an ant colony algorithm is improved to plan assembly sequences of the structural volume elements. Finally, a case illustrates the effectiveness of the approach.