Acoustic ecologist Bernie Krause hypothesized that rich soundscapes in mature ecosystems are generated by sound communication between different species with differentiating acoustic niches. This hypothesis, called the acoustic niche hypothesis, proposes that in a mature ecosystem, the singing of a species occupies a unique bandwidth in frequency and shifts in time to avoid competition, thus making the communication efficient. We hypothesize that selective pressure on communication complexity is required for differentiating and filling acoustic niches by a limited number of species, in addition to selective pressures on communication efficiency. To test this hypothesis, we built an evolutionary model where agents can emit complex sounds. Our simulations with the model demonstrate that selective pressure on communication efficiency and complexity leads to an evolution in spectral differentiation with a limited number of species filling the acoustic niche. This is the first demonstration of acoustic niche differentiation using an artificial life model with complex-sounding agents. We also propose multi-timescale complexity measurement, extending the Jensen–Shannon complexity using multi-scale permutation entropy. We analyze the evolved soundscape in the simulations using this measure. The result shows that multi-timescale complexity in soundscape evolved, suggesting that evolving niche differentiation leads to ecological complexity. We implement the extended model in real space and demonstrate that the system can adaptively generate sounds, differentiating acoustic niches with environmental sounds.