Periodic sweep-net sampling and capitula sampling were conducted between April and October 2007 to determine the seasonal phenology of Urophora quadrifasciata (Meigen) (Diptera: Tephritidae) on spotted knapweed, Centaurea stoebe subsp. micranthos (Gugler) Hayek (Asteraceae), in the Arkansas Ozarks of the United States of America. Urophora quadrifasciata produces up to three generations in Arkansas, rather than being bivoltine as in the northern United States of America. The first, second, and third generations of U. quadrifasciata reached peak adult numbers around 26 May, 29 June, and 27 July, respectively. Males tended to emerge earlier than females but the female:male ratio was approximately 1:1 for most of the season. The majority of the offspring of the three generations emerged between June and October, while approximately 38% entered diapause and emerged as adults in April–June of the following year; 3.4 ± 0.1 (mean ± SE) (range 1–12) flies emerged from each infested capitulum. The absence of other, competing knapweed biological control agents as well as very low rates of parasitism, mild weather conditions, and a longer knapweed growing season likely contributed to the adaptation and establishment of U. quadrifasciata on spotted knapweed in Arkansas.