Bispyribac-sodium, a herbicide that inhibits acetolactate synthase (ALS), is frequently used in rice fields in India to control weeds, including the most common noxious weed, barnyardgrass. However, rice growers have recently reported reduced control of barnyardgrass with bispyribac-sodium. Hence, a large-scale survey was carried out to assess bispyribac-sodium resistance in Chhattisgarh and Kerala, two rice-growing states. Open-field pot experiments were conducted for 2 yr to confirm resistance to bispyribac-sodium. Of the 37 biotypes tested, 30% (11) survived the recommended label rate of bispyribac-sodium (25 g ai ha−1). The effective rate of bispyribac-sodium required to achieve 50% control (ED50) of putative resistant biotypes ranged from 18 to 41 g ha−1, whereas it was about 10 g ha−1 for susceptible biotypes. This suggests that putative biotypes were two to four times more resistant to bispyribac-sodium. At 6 d after herbicide application, an in vitro enzyme assay demonstrated higher ALS enzyme activity in putative resistant biotypes (66% to 75%) compared with susceptible biotypes (48% to 52%). This indicates the presence of an insensitive ALS enzyme in those biotypes and a target site mutation as a possible mechanism for resistance. Whole-plant bioassays also suggested that the resistance problem is more widespread in Chhattisgarh than in Kerala. This study confirmed the first case of evolved resistance in barnyardgrass to bispyribac-sodium in rice fields of India.