The dark (non-photosynthetic) CO/sub 2/ fixation was studied in the leaves of ammonium-fed and nitrate-fed tomato plants. The ability to fix /sup 14/CO/sub 2/ in the dark of ammonium-fed plants was remarkably lower as compared with nitrate-fed plants, supporting the previous finding that the synthesis of C/sub 4/-compounds from C/sub 3/-compounds was reduced in the leaves of ammonium-fed plants. There was no difference in the activity of PEP carboxylase in extracts prepared from the leaves between both the plants during an early period of the treatment. However, the enzyme activity began to decrease rapidly in ammonium-fed plants 4 days after the treatment. By long-term treatments, the enzyme activity in ammonium-fed plants became half as high as that of nitrate-fed plants. The decreased PEP carboxylase activity in ammonium-fed plants was not associated with the presence of NH/sub 4/-N and the absence of NO/sub 3/-N in the leaf extract, and was not restored by the addition of the leaf extract from nitrate-fed plants. It is concluded that the decreased rate of synthesis of C/sub 4/-compounds from C/sub 3/-compounds in ammonium-fed plants is closely associated with a decrease in the dark fixation involving PEP carboxylase.