The effects of light and darkness on stomatal aperture and guard cell apoplastic pH in the intact leaf and in the isolated epidermal strips of Commelina communis have been investigated. Stomata in the intact leaf opened wide in the light. In contrast, sto

The walls of guard cells have many specialized features. Guard cells are present in the leaves of bryophytes, ferns, and almost all vascular plants. However, they exhibit considerable morphological diversities. There are two types of guard cells: the first type is found in a few monocots, such as palms and corn, and the other is found in most dicots, many monocots, mosses, ferns, and gymnosperms. In corns, guard cells have a characteristic dumbbell shape with bulbous ends. Most dicot and monocot species have kidney-shaped guard cells that have an elliptical contour with a pore at its center. Although subsidiary cells are common in species with kidney-shaped stomata, they are almost always absent in most of the other plants. In this study, there were many different stomatal features that were associated with kidney-shaped guard cells, but not dumbbell shaped guard cells, which are present in most grasses, such as cereals. Each plant investigated exhibited different characteristic features and most of these plants had kidney-shaped guard cells. However, the guard cells of Chamaesyce supina Mold, were often more rectangular than kidney-shaped. In contrast, Sedum sarmentosum guard cells were of the sink ensiform type and in Trifolium repens, the guard cells exhibited a more rhombic shape. Therefore, kidney-shaped guard cells could be divided into a number of subtypes that need to be investigated further.

The effect of CO2 on the opening of stomata in the intact leaf of Commelina communis has been investigated. Full opening of stomatal apertures(around 18 μm) was achieved in the intact leaf by addition of CO2(900 μmol mol-1). At 90 minutes, the stomatal apertures of leaves treated with CO2 free air were reduced. In contrast, stomata opened most widely with the treatment of CO2 air at 90 minutes. The effects of light, CO2 air and CO2 free air on the change of membrane potential difference(PD) were measured. Fast hyperpolarization of guard cell membrane PD was recorded reaching up to -12 mV in response to light. If CO2 free air was given firstly, there was no response. When light was given after CO2 free air, the light effect was very clear. At the onset of CO2 air, the PD showed a dramatic hyperpolarization to about -25 mV. Changes in the pH of apoplast in intact leaves in response to CO2 air were observed. CO2 air caused a change of 0.4 pH unit. Therefore, it can be hypothesized that CO2 flowing could stimulate proton efflux which is a necessary precursor of stomatal opening.

The plant hormone abscisic acid (ABA) serves as an integrator of environmental stress such as drought, to trigger stomatal closure by regulating specific ion channels in guard cells. We previously reported that SLAC1, an outward anion channel required for stomtal closure, was regulated via reversible protein phosphorylation events involving ABA signaling components including protein phosphatase 2C members and a SnRK2-type kinase (OST1). In this study, we reconstituted the ABA signaling pathway as a protein-protein interaction relay from the PYL/RCAR type receptors, to the PP2C-SnRK2 phosphatase-kinase pairs, to the ion channel SLAC1. The ABA receptors interact with and inhibit PP2C phosphatase activity against the SnRK2-type kinase, releasing active SnRK2 kinase to phosphorylate and activate the SLAC1 channel, leading to reduced guard cell turgor and stomatal closure. Both yeast-two hybrid and bi-molecular fluorescence complementation assays were used to verify the interactions among the components in the pathway. The biochemical assays demonstrated the activity modifications of phosphatases and kinases by their interaction partners. The SLAC1 channel activity was used as a readout for the strength of the signaling pathway depending on the presence of different combinations of signaling components.