Flowering and morphology of four Petunia Juss. spp. [P. axillaris (Lam.) Britton et al., P. exserta Stehmann, P. integrifolia (Hook.) Schinz & Thell., and P. ×hybrida Vilm.] were evaluated in response to photoperiod and temperature. Photoperiod responses were evaluated under 9-h short days (SD), 9-h photoperiod plus 4-h night-interruption lighting (NI), or a 16-h photoperiod supplemented with high-pressure sodium lamps (16-h HPS). All species flowered earlier under NI than SD and were classified as facultative (quantitative) long-day plants. Increasing the daily light integral within long-day treatments increased flower bud number for P. axillaris only. In a second experiment, crop timing and quality were evaluated in the temperature range of 14 to 26 °C under 16-h HPS. The rate of progress toward flowering for each species increased as temperature increased from 14 to 26 °C, suggesting the optimal temperature for development is at least 26 °C. The calculated base temperature for progress to flowering varied from 0.1 °C for P. exserta to 5.3 °C for P. integrifolia. Flowering of P. axillaris and P. integrifolia was delayed developmentally (i.e., increased node number below the first flower) at 14 °C and 17 °C or less, respectively, compared with higher temperatures. Petunia axillaris and P. integrifolia flower bud numbers decreased as temperature increased, whereas P. ×hybrida flower bud number was similar at all temperatures. The differences in crop timing and quality traits observed for these species suggest that they may be useful sources of variability for petunia breeding programs.