Macdunnoughia crassisigna Warren (Lepidoptera: Noctuidae) is a highly destructive herbivore that poses a serious risk to cotton, maize, soybean, and cruciferous vegetables in East Asia. Examining the effects of various biotic and abiotic factors on the flight performance of M. crassisigna is crucial for a better understanding of its trans-regional migration. In this study, the flight activity of M. crassisignai moths of different ages, under different temperatures and relative humidity (RH) levels, was evaluated by tethering individuals to computerized flight mills for a 24-h trial period. The results showed that M. crassisignai had the capacity for sustained flight and the flight ability was strongest in 3-day-old individuals, and then their flight performance decreased significantly in older moths. For both sexes, temperature had a significant effect on their flight performance, and the flight activity was relatively higher at 24–28°C than other temperatures. There was a significant effect of RH on all flight parameters of the tested moths, and the flight activity was relatively higher at RH of 60–75% than other RH levels. For 3-day-old moths under the optimum conditions (24°C and 75% RH) throughout the 24 h scotophase, their mean flight distance reached 66 km, and the mean flight duration reached 13.5 h, suggesting M. crassisigna possess strong potential to undertake long-distance migration. These findings will be helpful for developing sound forecasting systems of this pest species.