Wounding lettuce (Lactuca sativa L., Longifolia) leaves by excising 5-mm thick mid-rib segments increased phenylpropanoid metabolism with the synthesis and accumulation of wound-induced phenolic compounds (WIPC). Immersing freshly excised segments for 1 h in 20 degrees C water agitated with air or N-2 produced a 45% or 65% reduction in wound-induced phenolic content, respectively, compared to non-immersed segments when measured after incubation for 48 h at 10 degrees C in air. In contrast, agitating the water with O-2 produced a 23% increase in WIPC over the non-immersed controls. The enhanced reduction of WIPC in N-2 versus air agitated water, and the increase in WIPC in O-2 agitated water suggests that anaerobiosis, and not dilution of the wound signal, was the cause of the reduction in WIPC. Holding 5-mm segments in an anaerobic N-2 atmosphere produced a similar reduction in WIPC as did holding the segments in water. Delaying the 1 h anaerobic treatment for up to 3 h had no significant effect on the ability of the anaerobic treatment to reduce WIPC. Exposing 8-cm long midrib sections to anaerobiosis for 2 h before excision of the 5-mm segments reduced subsequent WIPC from the 5 mm segments. The previous anaerobic treatment of the 8-cm sections predisposed the tissue to have a reduced response to subsequent wounding. After a 2 d lag in WIPC accumulation, the rates of accumulation were similar for the air and 2 h anaerobic treated 5-mm segments. Using vacuum treatments to facilitate the loss of volatile products of anaerobic metabolism (e.g., acetaldehyde and ethanol) did not have a significant effect on the accumulation of WIPC. Ion leakage from the symplastic volume of the tissue (i.e., across the cell membrane) was unaffected by the anaerobic treatments, but leakage from the apoplastic volume increased with increasing duration of the anaerobic treatment. Immersing fresh-cut lettuce in an aqueous solutions did not reduce the wound response because of dilution of the wound signal, but because of the anaerobic environment created within the tissue. Some remnant of the anaerobic treatment seems to persist in the tissue and delay the accumulation of WIPC.