Abstract Non-native lady beetle species have often been introduced, with variable success, into North America for biological control of aphids, scales, whiteflies, and other agricultural pests. Two predatory lady beetle species, Propylea quatuordecimpunctata and Hippodamia variegata , both originating from Eurasia, were first discovered near Montreal, Quebec, in North America in 1968 and 1984, respectively, and have since expanded into northeastern North America and the midwestern United States. In this study, we estimate the range-wide population structure, establishment and range-expansion, and recent evolutionary history of these species of non-native lady beetles using reduced representation genotyping-by-sequencing via ddRADseq. In addition, we quantified the responses to a key abiotic factor, photoperiod, that regulates adult reproductive diapause in these two species and may influence their latitudinal distribution and spread in North America. Our analyses detect (1) non-significant genetic differentiation and divergence among North American populations, (2) evidence of reduced contemporary gene flow within the continental US, (3) significant phenotypic differences in diapause induction despite genetic similarities across sampled populations.