Osmoregulation in guard cells of intact, attached Vicia faba leaves gr
own under growth chamber and greenhouse conditions was studied over a
daily light cycle of stomatal movements. Under both growth conditions
guard cells had two distinct osmoregulatory phases. In the first (morn
ing) phase, opening was correlated with K+ uptake and, to a lesser ext
ent, sucrose accumulation. In the second (afternoon) phase, in which a
pertures were maximal, K+ content declined and sucrose became the domi
nant osmoticum. Reopening of the stomata after a CO2-induced closure w
as accompanied by accumulation of either K+ or sucrose, depending on t
he time of day, indicating that a single environmental signal may use
multiple osmoregulatory pathways. Malate accumulation, correlated with
K+ uptake, was detected under growth chamber but not greenhouse condi
tions, whereas Cl- was the main K+ counterion in the greenhouse. These
results indicate that guard-cell osmoregulation in the intact leaf de
pends on at least two different osmoregulatory pathways, K+ transport
and sucrose metabolism. Furthermore, the relative importance of the K counterions malate and Cl- appears to be environment-dependent.