Field and greenhouse experiments were conducted to determine the level of glyphosate resistance in common waterhemp populations from Platte County (MO1) and Holt County, Missouri (MO2), and to determine the level and distribution of resistance to glyphosate, acetolactate synthase (ALS)–inhibiting herbicides, and protoporophyrinogen oxidase (PPO)–inhibiting herbicides across the MO1 site. Results from greenhouse experiments revealed that the MO1 and MO2 waterhemp populations were 19 and 9 times more resistant to glyphosate, respectively, than a susceptible waterhemp population. In field experiments, greater than 54% of waterhemp at the MO1 site survived 1.7 kg glyphosate ae ha−1 (twice the labeled rate) 6 wk after treatment. Tank-mix combinations of ALS- and PPO-inhibiting herbicides with glyphosate also failed to provide complete control of the waterhemp population at the MO1 site. Collection and screening of seed from individual female waterhemp accessions revealed multiple resistance to glyphosate, ALS-, and PPO-inhibiting herbicides across the MO1 site. All 14 waterhemp accessions collected across the MO1 site exhibited greater than 65% survival to 2× rates of glyphosate and thifensulfuron, and these accessions were spread across a 5-km2 (503-ha) area. Four waterhemp accessions collected across a 0.9-km2 (87-ha) area also exhibited 26 to 38% survival to 2× rates of lactofen. The results from these experiments provide evidence and confirmation of the first glyphosate-resistant waterhemp population in the United States and reveal that multiple resistance to glyphosate, ALS-, and PPO-inhibiting herbicides can occur in waterhemp.