Enhanced herbicide metabolism is less common than target site–based herbicide resistance in weeds and often confers resistance to chemically dissimilar herbicides. In a previous study, the mechanism of acetolactate synthase (ALS)-inhibitor resistance in a downy brome biotype was determined to be metabolism. Our research was aimed at determining the multiple resistance pattern in the downy brome biotype, establishing its physiological basis, and investigating its fitness. Dose–response experiments showed that the resistant biotype was also moderately resistant to ethofumesate, clethodim, fluazifop, diuron, and terbacil and highly resistant to the triazine herbicides, atrazine and metribuzin. DNA sequence analysis of the psbA gene, which is the target site of PSII inhibitors, demonstrated a single amino acid substitution from serine to glycine in the resistant biotype at residue 264 in the D1 protein. Thus, the resistant biotype contains two different resistance mechanisms, herbicide metabolism and an altered target site. The resistant biotype produced less shoot dry weight, leaf area, and seed and was shorter than the susceptible biotype. The resistant biotype was also less competitive than the susceptible biotype. Thus, in the absence of herbicides, the frequency of this resistant biotype is unlikely to increase in a population of mixed downy brome biotypes.