ofGermany ABSTRACr A leaf cuvette has been adapted for use with a pulse-modulation fluorometer and an open gas exchange system. Leaf water potential (*) was decreased by withholding watering from Digitalis lanata EHRH. plants. At different stages of water deficiency the photochemical (qQ) and nomphotochemical (q,E) fluorescence quenching was determined during the transition between darkness and light-induced steady state photosynthesis of the attached leaves. In addition, the steady state CO2 and H20 gas exchange was recorded. Following a decrease of leaf water potential with increasing water deficiency, the transition of photochemical quenching was almost unaffected, whereas nonphotochemical quenching increased. This is indicative of an enhanced thylakoid membrane energization during the transition and is interpreted as a partial inhibition of either the ATP generating or the ATP consuming reaction sequences. Complete reversion of the stress induced changes was achieved within 6 hours after rewatering. In contrast to the variations during transition, the final steady state values of qQ and qE remained unchanged over the entire stress range from -0.7 to -2.5 megascals. From these results we conclude that, once established, electron transport via photosystem H and the transmembrane proton gradient remain unaffected by water stress. These data are indicative of a protective mechanism apinst photoinhibition during stress, when net CO2 uptake is limited.The relevant consequence of water stress is a reduction of growth, correlated to a diminished rate of net CO2 assimilation during the stress period (16). The fact that the intercellular CO2 concentration (C*)' does not decrease to the CO2 compensation point, but remains high even during moderate stress in several species (14,17, 29), invalidates the hypothesis that only stomatal closure is responsible for the loss of net photosynthesis via CO2 depletion (13).It has been established that the activity ofisolated chloroplasts is influenced by the osmotic potential of the isolation medium with respect to that of the extracted tissue (24). Therefore, previous measurements of electron transport and photophosphorylation from isolated organeiles should be interpreted with care. It seems to be useful to estimate electron transport ofslowly desiccated plants in vivo and to relate data on electron flow to ' Abbreviations: Ci, intercellular CO2 concentration; Fo, instantaneous fluorescence; F, peak fluorescence; Fm, maximal fluorescence; F, variable fluorescence at a given time; (F,)m, maximal variable fluorescence; (F,) It provides information about the onset kinetics of PCR activity, the rate ofelectron flow through PSII (photochemical quenching) and the energy status of the thylakoid membrane (energy-dependent, nonphotochemical quenching) (19, 27). In our system the fluorometer was connected to a small cuvette of an open gas exchange system. Fluorescence, net CO2 uptake and transpiration of the same leaf, which remained attached to the plant, were determined.
MATERIALS AND ME...