Opening Of Stomatal Pores Research Articles

Morpho-anatomical traits and leaf nutrient concentrations vary between plant communities in the Cerrado–Amazonia transition?

Leaves are the most variable plant organ, responding and adapting to the environment in the long and short term. However, the link between leaf functional traits and environmental gradients is challenging, especially in systems of high diversity, such as transitional regions between savannas and tropical forests that are considered unstable. Here we evaluate anatomical and morphological traits and nutrient concentrations for the most representative woody species of four vegetation types (i.e., semideciduous forest, cerradão, typical cerrado, and cerrado rupestre) in the Amazonia–Cerrado transition to explore how vegetation will respond to changes in the environment. We found that closed-canopy species invested in traits related to rapid resource acquisition (e.g., higher leaf water content, larger stomata size, and higher maximum opening of the stomatal pore), while open-canopy species optimized traits related to resource conservation and protection from physical damage (e.g., higher leaf thickness, smallest maximum stomatal pore opening and smaller size of stomata). Our study shows that functional traits play an important role in adapting to environmental gradients and help explain the performance of trees in the Amazonia–Cerrado transition, opening the door to a more accurate prediction of plant community responses over time.

The CIPK23 protein kinase represses SLAC1-type anion channels in Arabidopsis guard cells and stimulates stomatal opening.

Guard cells control the opening of stomatal pores in the leaf surface, with the use of a network of protein kinases and phosphatases. Loss of function of the CBL-interacting protein kinase 23 (CIPK23) was previously shown to decrease the stomatal conductance, but the molecular mechanisms underlying this response still need to be clarified. CIPK23 was specifically expressed in Arabidopsis guard cells, using an estrogen-inducible system. Stomatal movements were linked to changes in ion channel activity, determined with double-barreled intracellular electrodes in guard cells and with the two-electrode voltage clamp technique in Xenopus oocytes. Expression of the phosphomimetic variant CIPK23T190D enhanced stomatal opening, while the natural CIPK23 and a kinase-inactive CIPK23K60N variant did not affect stomatal movements. Overexpression of CIPK23T190D repressed the activity of S-type anion channels, while their steady-state activity was unchanged by CIPK23 and CIPK23K60N . We suggest that CIPK23 enhances the stomatal conductance at favorable growth conditions, via the regulation of several ion transport proteins in guard cells. The inhibition of SLAC1-type anion channels is an important facet of this response.

The evolution of plant proton pump regulation via the R domain may have facilitated plant terrestrialization

Plasma membrane (PM) H+-ATPases are the electrogenic proton pumps that export H+ from plant and fungal cells to acidify the surroundings and generate a membrane potential. Plant PM H+-ATPases are equipped with a C‑terminal autoinhibitory regulatory (R) domain of about 100 amino acid residues, which could not be identified in the PM H+-ATPases of green algae but appeared fully developed in immediate streptophyte algal predecessors of land plants. To explore the physiological significance of this domain, we created in vivo C-terminal truncations of autoinhibited PM H+‑ATPase2 (AHA2), one of the two major isoforms in the land plant Arabidopsis thaliana. As more residues were deleted, the mutant plants became progressively more efficient in proton extrusion, concomitant with increased expansion growth and nutrient uptake. However, as the hyperactivated AHA2 also contributed to stomatal pore opening, which provides an exit pathway for water and an entrance pathway for pests, the mutant plants were more susceptible to biotic and abiotic stresses, pathogen invasion and water loss, respectively. Taken together, our results demonstrate that pump regulation through the R domain is crucial for land plant fitness and by controlling growth and nutrient uptake might have been necessary already for the successful water-to-land transition of plants.

The green light gap: a window of opportunity for optogenetic control of stomatal movement.

Green plants are equipped with photoreceptors that are capable of sensing radiation in the ultraviolet-to-blue and the red-to-far-red parts of the light spectrum. However, plant cells are not particularly sensitive to green light (GL), and light which lies within this part of the spectrum does not efficiently trigger the opening of stomatal pores. Here, we discuss the current knowledge of stomatal responses to light, which are either provoked via photosynthetically active radiation or by specific blue light (BL) signaling pathways. The limited impact of GL on stomatal movements provides a unique option to use this light quality to control optogenetic tools. Recently, several of these tools have been optimized for use in plant biological research, either to control gene expression, or to provoke ion fluxes. Initial studies with the BL-activated potassium channel BLINK1 showed that this tool can speed up stomatal movements. Moreover, the GL-sensitive anion channel GtACR1 can induce stomatal closure, even at conditions that provoke stomatal opening in wild-type plants. Given that crop plants in controlled-environment agriculture and horticulture are often cultivated with artificial light sources (i.e. a combination of blue and red light from light-emitting diodes), GL signals can be used as a remote-control signal that controls stomatal transpiration and water consumption.

Exogenous trehalose ameliorates methyl viologen induced oxidative stress through regulation of stomatal pore opening and glutathione metabolism in tomato seedlings

Oxidative burst is the common thread linking all abiotic and biotic stressors. So in this study non-reducing disaccharide trehalose (Tre) has been examined for attenuation of oxidative stress, imposed through methyl viologen (MV) foliar spray in tomato seedlings. Initially the ameliorating property was ascertained by leaf disc senescence and stomatal pore opening assay, before carrying out the foliar spray on seedlings. Both these assays showed Tre mediated mitigation of MV induced oxidative stress. It was also observed that trehalose dose determines the extent of stomatal pore opening. MV treated seedlings showed more reactive oxygen species (ROS) accumulation compared to control, Tre and MV + Tre treated plants both histochemically and quantitatively. Damage due to ROS accumulation was documented by quantifying malondialdehye and lipoxygenase activity. Finally, the response of the tomato seedlings to oxidative stress was monitored by quantifying the antioxidant enzyme activities and non-enzymatic antioxidant content under stressed and unstressed conditions. Also transcript levels of glutathione synthetase (GS) and gamma-glutamylcysteine synthetase (γ-ECS), enzymes responsible for GSH synthesis; and transcription factors no apical meristem ATAF and cup-shaped cotyledon (NAC2) and dehydration-responsive element-binding (DREB) were found to increase on application of Tre. All of these experimentations showed better adaptability and survivability of the seedlings treated exogenously with both MV and Tre in comparison to those treated only with MV.

Stomatal Opening Involves Polar, Not Radial, Stiffening Of Guard Cells

SummaryIt has long been accepted that differential radial thickening of guard cells plays an important role in the turgor-driven shape changes required for stomatal pore opening to occur [1, 2, 3, 4]. This textbook description derives from an original interpretation of structure rather than measurement of mechanical properties. Here we show, using atomic force microscopy, that although mature guard cells display a radial gradient of stiffness, this is not present in immature guard cells, yet young stomata show a normal opening response. Finite element modeling supports the experimental observation that radial stiffening plays a very limited role in stomatal opening. In addition, our analysis reveals an unexpected stiffening of the polar regions of the stomata complexes, both in Arabidopsis and other plants, suggesting a widespread occurrence. Combined experimental data (analysis of guard cell wall epitopes and treatment of tissue with cell wall digesting enzymes, coupled with bioassay of guard cell function) plus modeling lead us to propose that polar stiffening reflects a mechanical, pectin-based pinning down of the guard cell ends, which restricts increase of stomatal complex length during opening. This is predicted to lead to an improved response sensitivity of stomatal aperture movement with respect to change of turgor pressure. Our results provide new insight into the mechanics of stomatal function, both negating an established view of the importance of radial thickening and providing evidence for a significant role for polar stiffening. Improved stomatal performance via altered cell-wall-mediated mechanics is likely to be of evolutionary and agronomic significance.

Noncanonical Function of a Small-Molecular Virulence Factor Coronatine against Plant Immunity: An In Vivo Raman Imaging Approach.

Coronatine (1), a small-molecular virulence factor produced by plant-pathogenic bacteria, promotes bacterial infection by inducing the opening of stomatal pores, the major route of bacterial entry into the plant, via the jasmonate-mediated COI1-JAZ signaling pathway. However, this pathway is also important for multiple plant functions, including defense against wounding by herbivorous insects. Thus, suppression of the COI1-JAZ signaling pathway to block bacterial infection would concomitantly impair plant defense against herbivorous wounding. Here, we report additional, COI1-JAZ-independent, action of 1 in Arabidopsis thaliana guard cells. First, we found that a stereoisomer of 1 regulates the movement of Arabidopsis guard cells without affecting COI1-JAZ signaling. Second, we found using alkyne-tagged Raman imaging (ATRI) that 1 is localized to the endoplasmic reticulum (ER) of living guard cells of Arabidopsis. The use of arc6 mutant lacking chloroplast formation was pivotal to circumvent the issue of autofluorescence during ATRI. These findings indicate that 1 has an ER-related action on Arabidopsis stomata that bypasses the COI1-JAZ signaling module. It may be possible to suppress the action of 1 on stomata without impairing plant defense responses against herbivores.

Absence of OsβCA1 causes a CO2 deficit and affects leaf photosynthesis and the stomatal response to CO2 in rice.

Plants always adjust the opening of stomatal pores to adapt to the environment, for example CO2 concentration ([CO2 ]), humidity and temperature. Low [CO2 ] will trigger the opening of stomatal pores to absorb extra CO2 . However, little is known about how CO2 supply affects the carbon fixation and opening of stomatal pores in rice. Here, a chloroplast-located gene coding for β-carbonic anhydrase (βCA) was found to be involved in carbon assimilation and the CO2 -mediated stomatal pore response in rice. OsβCA1 was constitutively expressed in all tissues and its transcripts were induced by high [CO2 ] in leaves. Both T-DNA mutant and RNA interference lines showed phenotypes of lower biomass and CA activities. Knockout of OsβCA1 obviously decreased photosynthetic capacity, as demonstrated by the increased CO2 compensation point and decreased light saturation point in the mutant, while knockout increased the opening ratio of stomatal pores and the rate of water loss. Moreover, the mutant showed a delayed response to low [CO2 ], and stomatal pores could not be closed to the same degree as those of wild type even though the stomatal pores could rapidly respond to high [CO2 ]. Genome-wide gene expression analysis via RNA sequencing demonstrated that the transcript abundance of genes related to Rubisco, photosystem compounds and the opening of stomatal pores was globally upregulated in the mutant. Taken together, the inadequate CO2 supply caused by the absence of OsβCA1 reduces photosynthetic efficiency, triggers the opening of stomatal pores and finally decreases their sensitivity to CO2 fluctuation.

Callose implication in stomatal opening and closure in the fern Asplenium nidus

The involvement of callose in the mechanism of stomatal pore opening and closing in the fern Asplenium nidus was investigated by examination of the pattern of callose deposition in open and closed stomata, and by examination of the effects of callose degradation and inhibition or induction of callose synthesis in stomatal movement. Callose was identified with aniline blue staining and a callose antibody and degraded via beta-1,3-D-glucanase. Callose synthesis was inhibited with 2-deoxy-D-glucose and induced by coumarin or dichlobenil. Stomatal pore opening and closing were assessed by estimation of the stomatal pore width. The open stomata entirely lacked callose, while the closed ones displayed distinct radial fibrillar callose arrays in the external periclinal walls. The latter displayed local bending at the region of callose deposition, a deformation that was absent in the open stomata. Both callose degradation and inhibition of callose synthesis reduced the stomatal ability to open in white light and close in darkness. By contrast, callose synthesis induction considerably improved stomatal pore opening and reduced stomatal closure in same conditions. The present data revealed that: during stomatal closure the external periclinal guard cell walls experience a strong mechanical stress, probably triggering callose synthesis; and that callose participates in stomatal movement.

Structural Adaptations in Overwintering Leaves of Thermonastic and Nonthermonastic Rhododendron Species

Evergreen rhododendrons (Rhododendron L.) are important woody landscape plants in many temperate zones. During winters, leaves of these plants frequently are exposed to a combination of cold temperatures, high radiation, and reduced photosynthetic activity, conditions that render them vulnerable to photooxidative damage. In addition, these plants are shallow-rooted and thus susceptible to leaf desiccation when soils are frozen. In this study, the potential adaptive significance of leaf morphology and anatomy in two contrasting Rhododendron species was investigated. R. catawbiense Michx. (native to eastern United States) exhibits thermonasty (leaf drooping and curling at subfreezing temperatures) and is more winter-hardy [leaf freezing tolerance (LT50) of containerized plants ≈–35 °C], whereas R. ponticum L. (native to central Asia) is less hardy (LT50 ≈–16 °C), and nonthermonastic. Thermonasty may function as a light and/or desiccation avoidance strategy in rhododendrons. Microscopic results revealed that R. ponticum has significantly thicker leaf blades but thinner cuticle than R. catawbiense. There is one layer of upper epidermis and three layers of palisade mesophyll in R. catawbiense compared with two distinct layers of upper epidermis and two layers of palisade mesophyll in R. ponticum. We suggest that the additional layer of upper epidermis in R. ponticum and thicker cuticle and extra palisade layer in R. catawbiense represent structural adaptations for reducing light injury in leaves and could serve a photoprotective function in winter when leaf photochemistry is generally sluggish. Results also indicate that although stomatal density of R. ponticum is higher than that of R. catawbiense leaves, the overall opening of stomatal pores per unit leaf area (an integrated value of stomatal density and pore size) is higher by approximately twofold in R. catawbiense, suggesting that R. catawbiense may be more prone to winter desiccation and that thermonasty may be a particularly beneficial trait in this species by serving as a desiccation-avoidance strategy in addition to a photoprotection role.

GUARD CELL SIGNAL TRANSDUCTION.

Guard cells surround stomatal pores in the epidermis of plant leaves and stems. Stomatal pore opening is essential for CO2 influx into leaves for photosynthetic carbon fixation. In exchange, plants lose over 95% of their water via transpiration to the atmosphere. Signal transduction mechanisms in guard cells integrate hormonal stimuli, light signals, water status, CO2, temperature, and other environmental conditions to modulate stomatal apertures for regulation of gas exchange and plant survival under diverse conditions. Stomatal guard cells have become a highly developed model system for characterizing early signal transduction mechanisms in plants and for elucidating how individual signaling mechanisms can interact within a network in a single cell. In this review we focus on recent advances in understanding signal transduction mechanisms in guard cells.

Probable involvement of cytoskeleton in stomatal-pore formation in Asplenium nidus L.

Stomatal-pore formation in the fernAsplenium nidus L. commences in postcytokinetic guard cells at the mid-region of the ventral wall, before the deposition of any cellulosic wall material on it, by the local movement of the adjacent plasmalemmata apart from each other. In this way a rudimentary “internal stomatal pore” is formed. At this stage the ventral wall exhibits an undulated appearance and gives a positive reaction to aniline blue. Detailed study of postcytokinetic guard cells by electron microscopy, as well as after tubulin immunolabeling and actin staining, shows that stomatal pore initiation coincides with the initiation of the organization of the anticlinal microtubule bundles along the middle of the ventral wall and the colocalization of actin filaments at the same sites. Afterwards, the stomatal pore broadens towards the periclinal walls, a phenomenon keeping pace with the further bundling of the cytoskeletal elements beneath the plasmalemmata lining the middle of the ventral wall. At this stage the anticlinal microtubule bundles lining the stomatal pore are very prominent. The above findings, as well as the fact that treatments with antimicrotubule drugs inhibit the “internal” stomatal-pore formation, denote that the cortical cytoskeleton lining the ventral wall and particularly the microtubules are involved in this process. Afterwards, distinct local wall thickenings are deposited at the sites of junction of the mid-region of the ventral wall with the periclinal walls as well as at the junctions of the polar ventral-wall ends with the external periclinal wall. Along the middle-lamella region of the former wall thickenings the fore- and rear-chambers of the stomatal pore are formed. The final stomatal-pore opening is achieved by disruption of the expanded thin median periclinal wall region inherited from the guard cell mother cell and of the overlying cuticle, which covers the stomatal pore externally and internally. At the same time the fore-chamber of the stomatal pore broadens by a schizogenous opening towards the polar ventral-wall ends. The observations show that the stomatal-pore formation inA. nidus is a unique process, which is probably restricted to ferns.

Plant growth hormones control voltage-dependent activity of anion channels in plasma membrane of guard cells

THE opening of stomatal pores in the epidermis of plant leaves is caused by an increase in turgor pressure of the guard cells as a result of the accumulation of potassium salts1,2. Although growth hormones have been shown to affect stomatal opening3, the transduction pathways by which growth regulators exert their effects on stomatal action are largely unknown. Here we report that auxins can elicit stomatal opening. These phytohormones modulate anion channels4,5 in the plasma membrane in what may be an initial step in regulated volume increase in guard cells. Our patch-clamp experiments demonstrate that auxins can directly interact with the extracellular face of the channel. As a result, its activation potential is shifted towards the resting potential of the cell to favour transient channel opening.