Determining the intra-specific variability of response to a given herbicide is important for monitoring the possible shifts in the sensitivity of weed populations. This study describes the responses of populations of Alisma plantago-aquatica, Cyperus difformis, and Schoenoplectus mucronatus from Italy, Greece, Portugal, and Spain to penoxsulam, an acetolactate synthase (ALS) inhibitor widely used in rice. To evaluate previously evolved resistance to ALS inhibitors, sensitivity to azimsulfuron and bensulfuron-methyl was assessed. Dose-response experiments with penoxsulam were performed in a greenhouse simulating paddy rice field conditions. Log-logistic dose-response curves were used to estimate the ED50, ED80, ED90 and GR50, GR80, and GR90. To calculate the average ED and GR and assess the intra-specific variability, an artificial resampling method was performed. Populations ALSPA 0364, 0365, 0469, 0470, 0471; SCPMU 0371, 0475, 0267; CYPDI 0013, 0431, 0432, 0433 appeared to be resistant to sulfonylureas, while a higher sensitivity to penoxsulam was observed, while populations ALSPA 0363, CYPDI 0223 and SCPMU 9719 proved to be cross-resistant. Regardless of species, ED90 of susceptible populations were below penoxsulam label dose (40 g ai ha−1) while they reached values higher than 320 g ai ha−1 for resistant populations. Average GR50 were generally lower than ED50. Sensitivity variability among susceptible populations is relatively low, allowing for discrimination between susceptible and resistant populations, and previously evolved resistance to sulfonylureas can influence sensitivity to penoxsulam.