Photoperiod is considered the universal zeitgeber, regulating physiological processes in numerous animals. However, for animals in light-restricted habitats (e.g. burrows or cavities), thermoperiod may be a more important cue. Our study tested this hypothesis in the alfalfa leafcutting bee, Megachile rotundata, which nests in cavities and undergoes development within a brood cell. We assessed the role of environmental cues (thermoperiod and photoperiod) on the process of adult emergence by examining: 1) if those cues direct circadian rhythms, 2) which cue is more dominant, and 3) how sensitive developing bees and emergence-ready adults are to cues. Although we found that 20% of light penetrates the brood cell, and bees respond to photoperiod by synchronizing emergence, thermoperiod is the dominant cue. When presented with a conflicting zeitgeber, bees entrained to the thermophase instead of the photophase. When temperature cues were removed, we observed free-running of emergence, indicating that underlying circadian mechanisms can be synchronized by daily fluctuations in temperature. We also found that emerging bees were highly sensitive to even small increases in temperature, entraining to a ramp speed of 0.33°C/hour. The response and sensitivity to temperature cues suggest that M. rotundata evolved a temperature-mediated clock to mediate emergence from light-restricted cavities.

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