Osmoregulation of leaf motor cells.

"Osmotic Motors"--the best-documented explanation for plant leaf movements--frequently reside in specialized motor leaf organs, pulvini. The movements result from dissimilar volume and turgor changes in two oppositely positioned parts of the pulvinus. This Osmotic Motor is powered by a plasma membrane proton ATPase, which drives KCl fluxes and, consequently, water, across the pulvinus into swelling cells and out of shrinking cells. Light signals and signals from the endogenous biological clock converge on the channels through which these fluxes occur. These channels and their regulatory pathways in the pulvinus are the topic of this review.

Blue light inactivates plasma membrane H(+)-ATPase in pulvinar motor cells of Phaseolus vulgaris L.

Unilateral blue light irradiation induces bending of pulvini of Phaseolus vulgaris towards the source of light. The pulvinar bending is caused by a decrease in turgor pressure of motor cells that are irradiated with blue light. Decrease in the turgor pressure is caused by the net efflux of K(+) and counter anions, accompanying membrane depolarization. In the present study the effect of blue light on the activity of plasma membrane H(+)-ATPase was studied in relation to the membrane depolarization. The activity of the plasma membrane H(+)-ATPase was measured using protoplast suspensions prepared from laminar pulvini from primary leaves. A pulse of blue light under continuous red light irradiation induced both a transient increase in the external pH and transient inhibition of the vanadate-sensitive ATPase. Continuous blue light irradiation under continuous red light irradiation induced both a sustained increase in the external pH and sustained inhibition of the vanadate-sensitive ATPase. These results show that blue light inhibits the activity of the plasma membrane H(+)-ATPase. Inactivation of the plasma membrane H(+)-ATPase supports the membrane depolarization induced by the blue light irradiation.

Intracellular localization of phytochrome in Robinia pseudoacacia pulvini.

The intracellular localization of phytochrome in the pulvini of Robinia pseudoacacia L. was analyzed by immunogold electron microscopy after red (R; 15 min) and far-red (FR; 5 min) irradiation 2 h after the beginning of the photoperiod. Screening of the available antibodies by immunoblotting demonstrated that none of the oat (Avena sativa L.) anti-phytochrome A (phy A) monoclonal antibodies) (MAbs) detected Robinia phytochrome. A putative Robinia phy A was detected by immunoblotting using a MAb to mustard (Sinapis alba L.) phy A (CP 2/9). No cross-reactivity was observed in blots probed with a MAb against Cucumis sativus L. phy B (mAT1). Ultrathin sections of LR White resin-embedded pulvini were immunolabelled with CP 2/9 MAb. The labelling was restricted to cortical cells and there was no evidence of labelling either in the vascular system or in the epidermis. The pattern of labelling was the same in both extensor and flexor cells irrespective of whether phytochrome was in the far-red-absorbing (Pfr) state or had reverted to the red-absorbing (Pr) form. Isolated labels and clusters of labels were randomly distributed throughout the cytoplasm. Gold particles were also found in the interior of nuclei, chloroplasts and mitochondria.

Blue-light-dependent osmoregulation in protoplasts of Phaseolus vulgaris Pulvini.

Blue light was found to induce shrinkage of the protoplasts isolated from first-leaf lamina pulvini of 18-day-old Phaseolus vulgaris. The response was transient following pulse stimulation, while it was sustainable during continuous stimulation. No apparent difference was found between flexor and extensor protoplasts. Protoplasts of the petiolar segment located close to the pulvinus showed no detectable response. In the plants used, the pulvinus was fully matured and the petiole was ceasing its elongation growth. When younger, 12-day-old, plants were used, however, the petiolar protoplasts did respond to blue light. The pulse-induced response was similar to that in pulvinar protoplasts, although the response to continuous stimulation was transient and differed from that in pulvinar protoplasts. No shrinkage was induced in pulvinar protoplasts when the far-red-light-absorbing form of phytochrome was absent for a period before blue-light stimulation, indicating that the blue-light responsiveness is strictly controlled by phytochrome. Inhibitors of anion channels and H(+)-ATPase abolished the shrinking response, supporting the view that protoplasts shrink by extruding ions. The response of pulvinar protoplasts is probably involved in the blue-light-induced, turgor-based movement of pulvini. The blue-light responding system in pulvini is suggested to have evolved from that functioning in other growing organs.