The delayed choice gedanken-experiment initially proposed by J A Wheeler reveals the nontrivial features of wave–particle duality in quantum world, that is, the choice of whether to observe the particle/wave character of a quantum system at a delayed time can affect its evolution history. In this work, the Wheeler’s delayed-choice experiment is conducted and tested in a new regime based on a Rydberg-ground level atomic Mach–Zehnder interferometer with optical coherent manipulation. Our quantum system exhibits an excellent single-excitation of the collective atomic state due to the Rydberg blockade effect, and the delayed choice of the interferometer configuration is realized by a quantum random number generator based on high-quality Rydberg single-photon source. Our experimental results demonstrate the Wheeler’s nontrivial time order inversion in this quantum system and confirm the Copenhagen interpretation of the physical reality of quantum state, which means the particle/wave character can only be ascribed to a quantum system when it was measured. This work is encouraging toward testing the foundations of quantum mechanics using the Rydberg atoms.