Under red light in ambient CO2 guard cells of faba bean (Vicia faba) fix CO2 and accumulate sucrose, causing stomata to open. We examined whether at [CO2] low enough to limit guard cell photosynthesis stomata would open when illuminated with red (R) or far-red (FR) light. After illumination with R or FR in buffered KCl solutions, net stomatal opening was c. 3 mum (R and FR) in air containing 210-225 mul l(-1) CO2 and was 5 mum (R) or 6.5 mum (FR) in air containing 40-50 mul l(-1) CO2. Opening was fully inhibited by 3-(3,4-dichlorophenyl)-1,1 dimethyl urea, the calmodulin antagonist W-7, the ser/thr kinase inhibitor ML-9, and sodium orthovanadate, but not by dithiothreitol, which inhibits formation of zeaxanthin, the blue light photoreceptor of guard cells. Stomatal opening was accompanied by K+ uptake and starch loss. Similar results were obtained when leaves were exposed to conditions designed to lower intercellular leaf [CO2]. These data suggest that the guard cell chloroplasts transduce reduced [CO2], activating stomatal opening through an ion uptake mechanism that depends on chloroplastic photosynthetic electron transport and that shares downstream components of the blue light signal transduction cascade.