Difenzoquat herbicide is used for post-emergence control of wild oat (Avena fatua L) in small grain crops. Its mechanism of action is not known, but appears to have both paraquat-like contact activity and systemic growth-inhibition activity. Experiments were conducted in vitro to compare the contact activity of difenzoquat and paraquat, to examine the contact activity of difenzoquat in difenzoquat-resistant (accession LCS) and -susceptible (accession SB 18) A fatua biotypes and -tolerant 'Ernest' and -susceptible 'Verde' hard red spring wheat (Triticum aestivum L) cultivars, and to investigate the role of differential DNA synthesis as a mechanism for difenzoquat resistance. Difenzoquat at 1 mM induced significant electrolyte leakage from resistant and susceptible A fatua leaf tissue in both light and dark, but paraquat at 1 mM induced electrolyte leakage only in light and not in dark. Difenzoquat at 1 mM under light induced more electrolyte leakage from resistant and susceptible A fatua than from tolerant 'Ernest' and susceptible 'Verde' wheat. Paraquat under light conditions induced more electrolyte leakage than difenzoquat, but no significant differences among A fatua accessions and wheat cultivars were detected. Difenzoquat under light did not decrease chlorophyll levels for either A fatua accessions or wheat cultivars, whereas paraquat decreased chlorophyll content of all four plant types. Inhibition of DNA synthesis was not significantly different between susceptible and resistant A fatua, but was significantly greater in susceptible 'Verde' than tolerant 'Ernest' wheat. These data suggest that difenzoquat and paraquat, have different modes of contact activity.