Research on the acquisition of spatial knowledge not only enriches our understanding of the theory of spatial knowledge representation but also creates practical value for the application of spatial knowledge. The aim of this study is to understand the impact of different learning methods on the acquisition of spatial knowledge, including the role of 2D maps, the difference between physical interaction and virtual interaction, and whether passive learning can replace active learning in virtual environments. One experiment was conducted, in which landmark knowledge and configurational knowledge were measured. Results indicate that 2D maps play a supporting role in acquiring both landmark knowledge and configurational knowledge. In addition, physical learning was associated with better spatial knowledge representation compared with virtual learning. An analysis of observational data in the third comparison found no significant difference between passive learning and active learning using virtual street view maps. However, with high-quality learning materials, passive learning can contribute to the acquisition of spatial knowledge more efficiently than active learning.