Reversal of blue light-stimulated stomatal opening by green light

Blue light-stimulated stomatal opening in detached epidermis of Vicia faba is reversed by green light. A 30 s green light pulse eliminated the transient opening stimulated by an immediately preceding blue light pulse. Opening was restored by a subsequent blue light pulse. An initial green light pulse did not alter the response to a subsequent blue light pulse. Reversal also occurred under continuous illumination, with or without a saturating red light background. The magnitude of the green light reversal depended on fluence rate, with full reversal observed at a green light fluence rate twice that of the blue light. Continuous green light given alone stimulated a slight stomatal opening, and had no effect on red light-stimulated opening. An action spectrum for the green light effect showed a maximum at 540 nm and minor peaks at 490 and 580 nm. This spectrum is similar to the action spectrum for blue light-stimulated stomatal opening, red-shifted by about 90 nm. The carotenoid zeaxanthin has been implicated as a photoreceptor for the stomatal blue light response. Blue/green reversibility might be explained by a pair of interconvertible zeaxanthin isomers, one absorbing in the blue and the other in the green, with the green absorbing form being the physiologically active one.     

Current theories for mechanism of stomatal opening: Influence of blue light; mesophyll cells,and sucrose

Since the late 1960s, researchers have observed that starch in the chloroplasts of the guard cells breaks down during the day and accumulates in the dark. Based on this, carbohydrates have historically been regarded as the primary osmotica modulating stomatal opening. However, the discovery of an important role for potassium uptake has led to the replacement of that starch-sugar hypothesis. Current research now focuses mainly on how K⁺ is transported in and out of cells when the stomata open or close. However, questions remain concerning photoreceptors, and the functioning of guard cell chloroplasts is still disputed. Coincidentally, some recent study results have again suggested that sucrose may play a major role in guard cell osmoregulation, thus supporting the original theory of starch-sugar involvement.     

Nitric oxide inhibits blue light-specific stomatal opening via abscisic acid signaling pathways in Vicia guard cells

Recent evidence suggests that nitric oxide (NO) acts as an intermediate of ABA signal transduction for stomatal closure. However, NO's effect on stomatal opening is poorly understood even though both opening and closing activities determine stomatal aperture. Here we show that NO inhibits stomatal opening specific to blue light, thereby stimulating stomatal closure. NO inhibited blue light-specific stomatal opening but not red light-induced opening. NO inhibited both blue light-induced H(+) pumping and H(+)-ATPase phosphorylation. The NO scavenger 2-carboxyphenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (c-PTIO) restored all these inhibitory effects. ABA and hydrogen peroxide (H(2)O(2)) inhibited all of these blue light-specific responses in a manner similar to NO. c-PTIO partially restored the ABA-induced inhibition of all of these opening responses but did not restore inhibition of the responses by H(2)O(2). ABA, H(2)O(2) and NO had slight inhibitory effects on the phosphorylation of phototropins, which are blue light receptors in guard cells. NO inhibited neither fusicoccin-induced H(+) pumping in guard cells nor H(+) transport by H(+)-ATPase in the isolated membranes. From these results, we conclude that both NO and H(2)O(2) inhibit blue light-induced activation of H(+)-ATPase by inhibiting the component(s) between phototropins and H(+)-ATPase in guard cells and stimulate stomatal closure by ABA.     

Gas exchange in paphiopedilum: lack of chloroplasts in guard cells correlates with low stomatal conductance

Net photosynthesis and stomatal conductance were measured in attached leaves of Paphiopedilum insigne. At 20°C and a vapor-pressure deficit of 0.5 kilopascal, both net photosynthesis and stomatal conductance were light-saturated below 0.2 millimole per square meter per second, a response typical of shade plants. The absolute values of photosynthetic rate and conductance however were remarkably low, presumably reflecting an adaptation to the low-light, limited-nutrient habitat characteristic of these orchids. The leaves also showed a vapor-pressure deficit response, with net photosynthesis and conductance varying over a 2-fold range between 0.3 and 1.6 kilopascals.

These results confirm that Paphiopedilum stomata are functional. The correlation between achlorophyllous guard cells and low conductance rates, however, singles them out as an exceptional biological system, exhibiting basic differences from typical stomata in higher plants. Available evidence showing that guard-cell chloroplasts are needed to sustain high conductance rates at moderate to high irradiances indicates that the genetic changes leading to the loss of chloroplast differentiation in Paphiopedilum guard cells were not deleterious because of the low conductance rates characteristic of this genus.

     

Evidence for phytochrome involvement in light-mediated stomatal movement in Phaseolus vulgaris L.

Observations made with primary leaves of Phaseolus vulgaris L. demonstrated that phytochrome modulates light-induced stomatal movement. Removal of the far-red-absorbing form of the pigment (Pfr) with far-red (FR) radiation decreased the time required by the stomata to reach maximal opening following a dark-to-light transition; this effect of FR was fully reversible with red. Removal of Pfr with FR also decreased the time required to reach maximal closure following a light-to-dark transition, and the rate of closure was dependent on the final irradiation treatment before darkness. No evidence was found for phytochrome involvement in determining stomatal aperture under constant conditions of either darkness of light.Abbreviations and symbols Chl chlorophyll - D darkness - FR far-red - phytochrome photostationary state - Pfr, Pr FR- and R-absorbing forms of phytochrome, respectively - R red     

Phytochrome involvement in stomatal movement in Pisum sativum, Vicia faba and Pelargonium sp.

Red and blue light triggered the opening of isolated stomata of Pisum sativum L. cv. Peleg Alvador, Vicia faba L. (unknown cultivar) and Pelargonium sp. The stimulatory effect of short irradiation with red or blue light was reversed by a subsequent short irradiation with far-red light. In Pisum the stimulatory effect of a continuous irradiation with red or blue light was also abolished by a concomitant far-red light. In leaf pieces of P. sativum blue light was more effective than red, but not in isolated guard cells. In the presence of mesophyll, DCMU inhibited stomatal opening in red light more than in blue, and thus increased the relative response to blue light. This was less evident in isolated guard cells.     

Phytochrome is not involved in the red-light-enhancement of the stomatal blue-light-response in wheat seedlings

The stomatal response to blue light (BL) in wheat seedlings ( Triticum aestivum L. cv. Starke II, Weibull) was enhanced by background red light (R). This enhancement was only slightly affected by the addition of background far-red light (FR). Under similar light treatments, the addition of FR induced a 43% transformation from the far-red-absorbing form towards the red-absorbing form of phytochrome from etiolated oat ( Avena sativa L. cv. Sol II), immobilized on phenyl-sepharose. Furthermore, the enhancement of the stomatal BL-response by 15 min R was not reversed by a subsequent irradiation with 5 min FR. It is concluded that the red-light-enhancement of the stomatal blue-light-response in wheat seedlings does not involve a change in the photostationary state of phytochrome.     

Reversal by green light of blue light-stimulated stomatal opening in intact, attached leaves of Arabidopsis operates only in the potassium-dependent, morning phase of movement

Green light reversal of blue light-stimulated stomatal opening was discovered in isolated stomata. The present study shows that the response also occurs in stomata from intact leaves. Arabidopsis thaliana plants were grown in a growth chamber under blue, red and green light. Removal of the green light opened the stomata and restoration of green light closed them to baseline values under experimental conditions that rule out a mesophyll-mediated effect. Assessment of the response to green light over a daily time course showed that the stomatal sensitivity to green light was observed only in the morning, which coincided with the use of potassium as a guard cell osmoticum. Sensitivity to green light was absent during the afternoon phase of stomatal movement, which was previously shown to be dominated by sucrose osmoregulation in Vicia faba. Hence, the shift away from potassium-based osmoregulation in guard cells is further postulated to entail a shift from blue light to photosynthesis as the primary component of the stomatal response to light. Stomata from intact leaves of the zeaxanthin-less, npq1 mutant of Arabidopsis failed to respond to the removal or restoration of green light in the growth chamber, or to short, high fluence pulses of blue or green light. These data confirm previous studies showing that npq1 stomata are devoid of a specific blue light response. In contrast, stomata from intact leaves of phot1 phot2 double mutant plants had a reduced but readily detectable response to the removal of green light and to blue and green pulses.     

Phytochrome-membrane interactions as a factor in stomatal opening

Red light enhances stomatal opening in Commelina communis L. This light effect is reversed by far-red irradiation. Pretreatment with filipin, which competitively inhibits phytochrome binding to membranes, also inhibits light-enhanced opening. The pretreatment with filipin is more inhibitory if preceded by red irradiation, than after far-red irradiation. Similar results are obtained with cycloheximide and low temperature, which retard phytochrome synthesis more than its degradation. This result may indicate an enhanced release of phytochrome in the Pfr form from binding sites rather than release of phytochrome in the Pr form. This points towards the possibility that phytochrome degradation and its release from binding sites are coupled.     

Effect of salinity on stomatal number,size and opening in barley Genotypes

Salinity had a varying effect on stomatal frequency, size and pore in barley (Hordeum vulgare L.). This variation in stomatal frequency, size and pore under salinity offers a scope of selecting cultivars with lesser increase in stomatal number to avoid excessive loss of water due to transpiration without effecting photosynthetic efficiency.     

Phytochrome modulation of blue light-induced chloroplast movements in Arabidopsis

Photometric analysis of chloroplast movements in various phytochrome (phy) mutants of Arabidopsis showed that phyA, B, and D are not required for chloroplast movements because blue light (BL)-dependent chloroplast migration still occurs in these mutants. However, mutants lacking phyA or phyB showed an enhanced response at fluence rates of BL above 10 micromol m-2 s-1. Overexpression of phyA or phyB resulted in an enhancement of the low-light response. Analysis of chloroplast movements within the range of BL intensities in which the transition between the low- and high-light responses occur (1.5-15 micromol m-2 s-1) revealed a transient increase in light transmittance through leaves, indicative of the high-light response, followed by a decrease in transmittance to a value below that measured before the BL treatment, indicative of the low-light response. A biphasic response was not observed for phyABD leaves exposed to the same fluence rate of BL, suggesting that phys play a role in modulating the transition between the low- and high-light chloroplast movement responses of Arabidopsis.     

Environmental effects on circadian rhythms in photosynthesis and stomatal opening

Persistent circadian rhythms in photosynthesis and stomatal opening occurred in bean (Phaseolus vulgaris L.) plants transferred from a natural photoperiod to a variety of constant conditions. Photosynthesis, measured as carbon assimilation, and stomatal opening, as conductance to water vapor, oscillated with a freerunning period close to 24 h under constant moderate light, as well as under light-limiting and CO₂-limiting conditions. The rhythms damped under constant conditions conducive to high photosynthetic rates, as did rates of carbon assimilation and stomatal conductance, and this damping correlated with the accumulation of carbohydrate. No rhythm in respiration occurred in plants transferred to constant darkness, and the rhythm in stomatal opening damped rapidly in constant darkness. Damping of rhythms also occurred in leaflets exposed to constant light and CO₂-free air, demonstrating that active photosynthesis and not simply light was necessary for sustained expression of these rhythms. This is CIWDPB Publication No. 1142 This research was supported by National Science Foundation grant BSR 8717422 (C.B.F.) and a U.S. Department of Agriculture training grant to Stanford University (T.L.H.).     

Suppression of the stomatal opening by morphactins in isolated epidermal strips

The effects of three morphactins, chlorflurenol, flurenol andEMD 7301 W, were examined on the stomatal aperture using isolatedepidermal strips of Commelina benghalensis. Morphactins produced,a striking decrease in the stomatal opening in light but hadno effect on stomatal closure in darkness. Various catalystsand inhibitors of photophosphorylation had no influence on themorphactin-induced stomatal closure. The stimulatory effectsof ATP, pyruvate and KC1 on stomatal opening were suppressedby the morphactins. The cytokinin, benzyladenine stimulatedthe stomatal opening even in the presence of a morphactin. Theinfluence of morphactins on the stomatal aperture closely resembledthe effect of abscisic acid. ¹Present address: Central Plantation Crops Research Institute,Regional Station, Vittal 574243, Karnataka, India. (Received September 16, 1975; )     

The role of ion channels in light-dependent stomatal opening

Stomatal opening represents a major determinant of plant productivity and stress management. Because plants lose water essentially through open stomata, volume control of the pore-forming guard cells represents a key step in the regulation of plant water status. These sensory cells are able to integrate various signals such as light, auxin, abscisic acid, and CO(2). Following signal perception, changes in membrane potential and activity of ion transporters finally lead to the accumulation of potassium salts and turgor pressure formation. This review analyses recent progress in molecular aspects of ion channel regulation and suggests how these developments impact on our understanding of light- and auxin-dependent stomatal action.     

Role of vacuolar invertase in regulating Arabidopsis stomatal opening

Guard cells sense and integrate environmental signals to modulate stomatal aperture in response to diverse conditions. In this study, the effect of vacuolar invertase on Arabidopsis stomatal opening was investigated. The technology of enzyme activity detection in situ was used to show that the vacuolar invertase activity was much higher in guard cells than in other epidermal cells. The stomatal aperture of T-DNA insertion mutant in At1g12240 (inv-7) was significantly lower than that in wild-type plants. Increased stomatal aperture was observed in the transgenic Arabidopsis overexpressing cotton vacuolar invertase gene. These results indicated that Arabidopsis stomatal aperture was correlated with vacuolar invertase, and that vacuolar invertase may play an important role in regulating Arabidopsis stomatal opening.     

Phototropin signaling and stomatal opening as a model case

Phototropins are plant-specific light-activated receptor kinases that regulate diverse blue-light-induced responses, and serve to optimize plant growth under various light environments. Phototropins undergo autophosphorylation as an essential step for their signaling and induce a variety of tissue-specific or organ-specific responses, but the divergent mechanisms for these responses are unknown. It is most likely that the phototropins generate a specific output after the event of autophosphorylation. In this report, we will review the common steps of phototropin signaling and the numerous interactive proteins of phototropins, which may act as signal transducers for the diverse responses. We also describe the phototropin-mediated signaling process of stomatal guard cells and its crosstalk with abscisic acid signaling.     

Phytochrome B control of total leaf area and stomatal density affects drought tolerance in rice

We report that phytochrome B (phyB) mutants exhibit improved drought tolerance compared to wild type (WT) rice (Oryza sativa L. cv. Nipponbare). To understand the underlying mechanism by which phyB regulates drought tolerance, we analyzed root growth and water loss from the leaves of phyB mutants. The root system showed no significant difference between the phyB mutants and WT, suggesting that improved drought tolerance has little relation to root growth. However, phyB mutants exhibited reduced total leaf area per plant, which was probably due to a reduction in the total number of cells per leaf caused by enhanced expression of Orysa;KRP1 and Orysa;KRP4 (encoding inhibitors of cyclin-dependent kinase complex activity) in the phyB mutants. In addition, the developed leaves of phyB mutants displayed larger epidermal cells than WT leaves, resulting in reduced stomatal density. phyB deficiency promoted the expression of both putative ERECTA family genes and EXPANSIN family genes involved in cell expansion in leaves, thus causing greater epidermal cell expansion in the phyB mutants. Reduced stomatal density resulted in reduced transpiration per unit leaf area in the phyB mutants. Considering all these findings, we propose that phyB deficiency causes both reduced total leaf area and reduced transpiration per unit leaf area, which explains the reduced water loss and improved drought tolerance of phyB mutants.