Stomatal Opening Research Articles

Dual function of overexpressing plasma membrane H+-ATPase in balancing carbon-water use.

Stomata respond slowly to changes in light when compared with photosynthesis, undermining plant water-use efficiency (WUE). We know much about stomatal mechanics, yet efforts to accelerate stomatal responsiveness have been limited despite the breadth of potential targets for manipulation. Here, we use mechanistic modeling to establish a hierarchy of putative targets affecting stomatal kinetics. Counterintuitively, modeling predicted that overexpressing plasma membrane H+-ATPases could speed stomata and enhance WUE under fluctuating light, even though overexpressed H+-ATPases is known to promote stomatal opening and reduce WUE in the steady state. Experiments validated the prediction, implicating an unexpected role of the H+-ATPases in improving WUE under fluctuating light. It suggests that H+-ATPases have a dual function, acting as a facilitator of carbon assimilation and water use, depending on the light conditions. These findings highlight the importance of integrating in silico modeling with experiments in future efforts toward enhancing stomatal function.

Deep Learning for Stomatal Opening Recognition in Gynura formosana Kitam Leaves

Gynura formosana Kitam possesses beneficial properties such as heat-clearing, detoxification, and cough suppression, making it a highly nutritious plant with significant economic value. During its growth, the plant’s leaves are prone to infections that can impair stomatal function and hinder growth. Effective identification of stomatal openings and timely application of appropriate chemicals or hormones or indirect environmental adjustments (such as light, temperature, and humidity) to regulate stomatal openings are essential for maintaining the plant’s healthy growth. Currently, manual observation is the predominant method for monitoring stomatal openings of Gynura formosana Kitam, which is complex, labor-intensive, and unsuitable for automated detection. To address this, the study improves upon YOLOv8s by proposing a real-time, high-precision stomatal detection model, Refined GIoU. This model substitutes the original IoU evaluation methods in YOLOv8s with GIoU, DIoU, and EIoU while incorporating the SE (Squeeze-and-Excitation) and SA (Self-Attention) attention mechanisms to enhance understanding of feature representation and spatial relationships. Additionally, enhancements to the P2 layer improve the feature extraction and scale adaptation. The effectiveness of the Refined GIoU is demonstrated through training and validation on a dataset of 1500 images of Gynura formosana Kitam stomata. The results show that the Refined GIoU achieved an average precision (mAP) of 0.935, a recall of 0.98, and an F1-score of 0.88, reflecting an excellent overall performance. The GIoU loss function is better suited to detecting stomatal openings of Gynura formosana Kitam, significantly enhancing the detection accuracy. This model facilitates the automated, real-time monitoring of stomatal openings, allowing for timely control measures and improved economic benefits of Gynura formosana Kitam cultivation.

Probing ozone effects on European hornbeam (Carpinus betulus L. and Ostrya carpinifolia Scop.) leaf water content through THz imaging and dynamic stomatal response

We investigated the impact of ozone exposure on Hornbeam using a novel dual approach based on Terahertz (THz) imaging in a free-air ozone exposure experiment (three ozone levels: ambient; 1.5 times ambient; twice ambient). The research aims at unraveling the physiological responses induced by elevated ozone levels on water dynamics. THz imaging unveiled dynamic changes in leaf water content, providing a non-invasive approach to leaf water monitoring. Leaf gas exchange measurements assessed stomatal responses to light variation. Our findings showcase a compelling correlation between elevated ozone levels and reduction in photosynthetic rate and impairment of stomatal function, i.e. “stomatal sluggishness”, indicative of nuanced regulatory mechanism. Stomatal sluggishness was particularly evident in Carpinus betulus (CB) compared to Ostrya carpinifolia (OC) and was linked to reduction in photosynthetic capacity. THz-based imaging techniques confirmed this result indicating a negative effect of O3 on leaf-level total water content. In addition, spatial analysis of leaf water status using these techniques also highlighted that the negative effect of O3 on water status was progressing even in less sensitive OC plants though visible foliar injury was not detected. In fact, OC showed a relative dry area of 1.6 ± 1.6 % in the control group and 3.8 ± 1.3 % under high ozone levels. THz-based imaging techniques provided a deep understanding of O3 behavior in plants and may be recommended for precision biosensing in the early detection of O3-induced damage. The integration of THz imaging and physiological analysis resulted in comprehensive understanding of Hornbeam acclimation response to ozone exposure.

Guard Cell Activity of PIF4 Represses Disease Resistance in Arabidopsis.

Phytochrome Interacting Factor 4 (PIF4) plays a central role in coordinating plant growth regulation by integrating multiple environmental cues. However, studies on whether and how PIF4 regulates plant immunity have inconsistent findings. In this study, we investigated the role of PIF4 in disease resistance against Pst DC3000 by characterizing its loss-of-function mutants using different inoculation strategies. Our findings reveal that pif4 mutants exhibit enhanced disease resistance with spray inoculation but not with infiltration inoculation compared to wild-type plants, and that mutants displayed more closed stomata apertures, indicating that PIF4 promotes stomatal opening. Importantly, expression of PIF4 by a guard-cell-specific promoter was sufficient to restore disease resistance to the wild-type level in the pif4 mutant. Additionally, PIF4 overexpression enhances disease symptom development independent of disease resistance and chlorophyll degradation, while the loss of PIF4 function leads to higher chlorophyll accumulation. Thus, our findings highlight a crucial function of PIF4 in regulating stomata-mediated disease resistance and chlorophyll accumulation, providing new insights into the connection of growth and defense in plants.

Subfamily C7 Raf-like kinases MRK1, RAF26, and RAF39 regulate immune homeostasis and stomatal opening in Arabidopsis thaliana.

The calcium-dependent protein kinase CPK28 regulates several stress pathways in multiple plant species. Here, we aimed to discover CPK28-associated proteins in Arabidopsis thaliana. We used affinity-based proteomics and identified several potential CPK28 binding partners, including the C7 Raf-like kinases MRK1, RAF26, and RAF39. We used biochemistry, genetics, and physiological assays to gain insight into their function. We define redundant roles for these kinases in stomatal opening, immune-triggered reactive oxygen species (ROS) production, and resistance to a bacterial pathogen. We report that CPK28 associates with and trans-phosphorylates RAF26 and RAF39, and that MRK1, RAF26, and RAF39 are active kinases that localize to endomembranes. Although Raf-like kinases share some features with mitogen-activated protein kinase kinase kinases (MKKKs), we found that MRK1, RAF26, and RAF39 are unable to trans-phosphorylate any of the 10 Arabidopsis mitogen-activated protein kinase kinases (MKKs). Overall, our study suggests that C7 Raf-like kinases associate with and are phosphorylated by CPK28, function redundantly in stomatal opening and immunity, and possess substrate specificities distinct from canonical MKKKs.

Melatonin Rinsing Treatment Associated with Storage in a Controlled Atmosphere Improves the Antioxidant Capacity and Overall Quality of Lemons.

Antioxidant capacity is one of the most important biological activities in fruits and vegetables and is closely related to human health. In this study, 'Eureka' lemons were used as experimental materials and stored at 7-8 °C MT (melatonin, 200 μmol, soaked for 15 min) and CA (controlled atmosphere, 2-3% O2 + 15-16% CO2) individually or in combination for 30 d. The changes in lemon fruits' basic physicochemical properties, enzyme activities, and antioxidant capacities were studied. Comparing the combined treatment to the control, the outcomes demonstrated a significant reduction in weight loss, firmness, stomatal opening, and inhibition of polyphenol oxidase (PPO) and peroxidase (POD) activities. Additionally, the combined treatment maintained high levels of titratable acidity (TA), vitamin C (VC), total phenolic content (TPC), and antioxidant capacity and preserved the lemon aroma. Meanwhile, the correlation between fruit color, aroma compounds, and antioxidant capacity was revealed, providing valuable insights into the postharvest preservation of lemons. In conclusion, the combined treatment (MT + CA) was effective in maintaining the quality and antioxidant capacity of lemons.

Optimizing micropropagation of Miscanthus sinensis ‘Gold Bar’ by shortening the production cycle and reducing acclimation stress through the use of selected growth regulators

Miscanthus, with its decorative qualities and low cultivation requirements in terms of soil fertility and temperature, is the most popular grass in gardens and urban areas. For years, micropropagation has been regarded as an effective method of its production. However, in order to meet the demands of customers and provide adequate quantities of high-quality planting materials, it is necessary to develop more efficient methods of Miscanthus production. The present study evaluated the influence of different media on Miscanthus sinensis (Thunb.) Andersson (silver grass) multiplication as well as assessing the effect of different concentrations of selected cytokinins and auxins on multiplication and rhizogenesis. To shorten the production cycle and reduce costs, ex vitro rooting was combined with acclimatization, and selected growth regulators were used to decrease stress associated with external conditions. Biochemical analyses were conducted at each stage to determine the content of basic organic compounds, hydrogen peroxide (H2O2) and catalase activity. Stomatal function was assessed at the acclimatization stage. The obtained results allowed for the production cycle of plants propagated in tissue culture to be shortened by simultaneous rooting and acclimatization of microcuttings sprayed with abscisic acid (ABA). This regulator has been shown to effectively reduce plant stress associated with acclimatization by reducing H2O2 concentration and increasing assimilation pigment content. Growth regulators reduced the number of stomata that developed on the leaves of silver grass and led to lower stomatal conductance. H2O2 should be considered not only a stress marker but a vital signaling molecule.

How the vertical gradient of light in the understorey and water seasonality affect leaf traits of Vanilla phaeantha (Orchidaceae), a crassulacean acid metabolism (CAM) hemiephyte.

Structural and physiological leaf traits and their plasticity were compared in the hemiepiphyte Vanilla phaeantha . This species grows along a phorophyte reaching different understorey positions and exhibiting diverse responses to environment changes. We analysed three height strata above the ground, establishing a light gradient, and considering seasonal water fluctuations. The upper leaves had higher area and mass and were less pigmented. The dry season induced a reduction of approximately 2h of stomatal opening over the diel 24h crassulacean acid metabolism (CAM) cycle in the leaves at all understorey positions. The leaves more exposed to sunlight were larger with higher titratable acidity during the rainy season, while the leaves near the ground maintained the same rates of stomatal conductance and nocturnal acidification between seasons, with lowest values of carbon isotopes in the rainy season. Our research showed that some structural leaf traits (such as specific leaf mass, biomass, and saturated water content) are sensitive to variation in understorey position. In contrast, other physiological traits (stomatal conductance, transpiration, and fluorescence parameters) are more sensitive to seasonal variations. The results are a novelty in assessing the variation of CAM along the same plant in a height gradient and under field conditions.

Effects of mineral fertilization (NPK) on combined high temperature and ozone damage in rice

BackgroundIncreasing concern has recently been highlighted regarding crop damage due to extreme weather events caused by global warming and the increased production of ground-level ozone. Several studies have investigated rice growth in response to fertilization conditions under various environmental stress conditions; however, studies on growth development in response to fertilization conditions under combined high-temperature/ozone treatment conditions are scarce. In this study, we aimed investigate the growth and physiological development of rice under combined high temperature and ozone treatment conditions and to reveal the damage-mitigation effects of NPK fertilization treatments.ResultsThe plants were treated with varying levels of NPK [N2 (N-P-K: 9.0-4.5-4.0 kg/a), P2 (4.5-9.0-4.0 kg/a), K2 (4.5-4.5-8.0 kg/a), and control (4.5-4.5-4.0 kg/10a).] under combined high-temperature (35 ℃) and ozone (150 pb) treatment conditions. Analysis of the growth metrics, including plant height, leaf age, dry weight, and the plant height/leaf age (PH/L) ratio were revealed that combined high-temperature/ozone treatment promoted the phenological development indicated by increasing leaf age but decreased the plant height and dry weight indicating its negative effect on quantitative growth. The effects of this combined high-temperature/ozone treatment on growth were alleviated by NPK fertilization, particularly in K2 treatment but worsened in N2 treatment. Visible damage symptoms in rice leaves induced by exposure to the combined stressors was also alleviated by the K2 treatment. At the physiological level, K2 treatment reduced the expression of OsF3H2, which is associated with antioxidant activity, suggesting that potassium improved stress tolerance. Additionally, expression of genes related to abscisic acid (ABA) metabolism showed increased OsNECD (ABA synthesis) and decreased OsCYP707A3 (ABA degradation) in the K2 treatment, promoting a stronger adaptive stress response. Stomatal conductance measurements indicated a slight increase under K2 treatment, reflecting enhanced regulation of stomatal function during stress.ConclusionThe study highlights the potential of potassium fertilization to mitigate combined high-temperature and ozone stress in rice, suggesting it as a strategy to improve crop resilience and optimize fertilization. The findings offer insights into fertilization treatments and can guide future research on stress tolerance in crops.

Wind speed affects the rate and kinetics of stomatal conductance.

Understanding the relationship between wind speed and gas exchange in plants is a longstanding challenge. Our aim was to investigate the impact of wind speed on maximum rates of gas exchange and the kinetics of stomatal responses. We conducted experiments in different angiosperm and fern species using an infrared gas analyzer equipped with a controlled leaf fan, enabling precise control of the boundary layer conductance. We first showed that the chamber was adequately mixed even at extremely low wind speed (<0.005 m s-1) and evaluated the link between fan speed, wind speed, and boundary layer conductance. We observed that higher wind speeds led to increased gas exchange of both water vapor and CO₂, primarily due to the increase in boundary layer conductance. This increase in transpiration subsequently reduced epidermal pressure, leading to stomatal opening. We documented that stomatal opening in response to light was 2.5 times faster at a wind speed of 2 m s-1 compared to minimal wind speed in Vicia faba, while epidermal peels in a buffer with no transpiration exhibited a similar opening rate. The increase in stomatal conductance under high wind was also observed in four angiosperm species under field conditions, but it was not observed in Boston fern (Nephrolepis exaltata), which lacks epidermal mechanical advantage. Our findings highlight the significant impact of boundary layer conductance on determining gas exchange rates and the kinetics of gas exchange responses to environmental changes.

OsCNGC7 modulates calcium dynamics and accelerates leaf senescence in rice

Calcium plays a crucial role in regulating plant senescence. However, the specific effects of increased intranuclear calcium versus cytoplasmic calcium on aging remain unclear. Cyclic nucleotide-gated channels (CNGCs), which manage Ca2⁺ levels in plant cells, are particularly significant in this context. These channels are known to relocate between the nuclear envelope and the plasma membrane in response to stress and developmental signals. Through this movement, CNGCs help regulate the balance of cytosolic and intranuclear Ca2⁺. In this study, we categorized the 16 CNGC genes in rice into five subgroups. OsCNGCs are notably expressed in leaves, especially during the reproductive stage. Both OsCNGC6 and OsCNGC7 exhibit dual localization to the plasma membrane and the nuclear envelope. Knockdown of OsCNGC7 led to reduced levels of Ca2⁺ and K⁺ in plants. Conversely, yeast expressing the OsCNGC7 gene showed increased sensitivity to Ca2⁺. Additionally, while the [Ca2⁺]cyt was maintained at relatively low levels in both wild-type and OsCNGC7-RNAi lines, the fluorescence intensity was significantly higher in OsCNGC7-overexpressing lines, particularly in the nucleus of root tips. Overexpression of OsCNGC7 resulted in enhanced stomatal opening and accelerated leaf senescence from the tillering stage to grain filling in rice. Treatment with MeJA rapidly induced OsCNGC7 expression, while knockdown of OsCNGC7 delayed both MeJA-induced and dark-induced leaf senescence. Further analysis revealed that OsCNGC7 interacts with OsKAT2 and OsALMT2. In conclusion, our findings highlight the distinct roles of OsCNGCs in regulating senescence. This knowledge could provide new strategies for manipulating plant senescence and enhancing crop productivity.

Fungal toxin fusicoccin enhances plant growth by upregulating 14-3-3 interaction with plasma membrane H+-ATPase.

Fusicoccin-A (FC-A) is a diterpene glucoside produced by a pathogenic fungus. Since its discovery, FC-A has been widely recognized as a phytotoxin that induces stomatal opening and leaf wilting, eventually leading to plant death. In this study, we present the first evidence that FC-A enhances plant growth by stabilizing the protein-protein interaction between plasma membrane (PM) H+-ATPase and 14-3-3 in guard cells. Long-term treatment of Arabidopsis plants with FC-A resulted in ~ 30% growth enhancement. Structurally similar fusicoccin-J (FC-J) showed a similar degree of growth-promotion activity as FC-A, whereas the more hydrophilic fusicoccin-H (FC-H) exhibited no effect on plant growth, indicating that the enhancement of plant growth observed with FC-A and FC-J involves upregulation of the protein-protein interaction between PM H+-ATPase and 14-3-3 in guard cells, which promotes stomatal opening and photosynthesis.

Metabolic modelling reveals distinct roles of sugars and carboxylic acids in stomatal opening and uncovers unexpected carbon fluxes.

Guard cell metabolism is crucial for stomatal dynamics, but a full understanding of its role is hampered by experimental limitations and the flexible nature of the metabolic network. To tackle this challenge, we constructed a time-resolved stoichiometric model of guard cell metabolism that accounts for energy and osmolyte requirements and which is integrated with the mesophyll. The model resolved distinct roles for starch, sugars, and malate in guard cell metabolism and revealed several unexpected flux patterns in central metabolism. During blue light-mediated stomatal opening, starch breakdown was the most efficient way to generate osmolytes with downregulation of glycolysis allowing starch-derived glucose to accumulate as a cytosolic osmolyte. Maltose couldalso accumulate as a cytosolic osmoticum, although this made the metabolic system marginally less efficient. The metabolic energy for stomatal opening was predicted to be derived independently of starch, using nocturnally accumulated citrate which was metabolised in the tricarboxylic acid cycle to malate to provide mitochondrial reducing power for ATP synthesis. In white light-mediated stomatal opening, malate transferred reducing equivalents from guard cell photosynthesis to mitochondria for ATP production. Depending on the capacity for guard cell photosynthesis, glycolysis showed little flux during the day but was crucial for energy metabolism at night. In summary, our analyses have corroborated recent findings in Arabidopsis guard cell research, resolved conflicting observations by highlighting the flexibility of guard cell metabolism, and proposed new metabolic flux modes for further experimental testing.

Warming triggers stomatal opening by enhancement ofphotosynthesis and ensuing guard cell CO2 sensing,whereashigher temperatures induce a photosynthesis-uncoupled response.

Plants integrate environmental stimuli to optimize photosynthesis vs water loss by controlling stomatal apertures. However, stomatal responses to temperature elevation and the underlying molecular genetic mechanisms remain less studied. We developed an approach for clamping leaf-to-air vapor pressure difference (VPDleaf) to fixed values, and recorded robust reversible warming-induced stomatal opening in intact plants. We analyzed stomatal temperature responses of mutants impaired in guard cell signaling pathways for blue light, abscisic acid (ABA), CO2, and the temperature-sensitive proteins, Phytochrome B (phyB) and EARLY-FLOWERING-3 (ELF3). We confirmed that phot1-5/phot2-1 leaves lacking blue-light photoreceptors showed partially reduced warming-induced stomatal opening. Furthermore, ABA-biosynthesis, phyB, and ELF3 were not essential for the stomatal warming response. Strikingly, Arabidopsis (dicot) and Brachypodium distachyon (monocot) mutants lacking guard cell CO2 sensors and signaling mechanisms, including ht1, mpk12/mpk4-gc, and cbc1/cbc2 abolished the stomatal warming response, suggesting a conserved mechanism across diverse plant lineages. Moreover, warming rapidly stimulated photosynthesis, resulting in a reduction in intercellular (CO2). Interestingly, further enhancing heat stress caused stomatal opening uncoupled from photosynthesis. We provide genetic and physiological evidence that the stomatal warming response is triggered by increased CO2 assimilation and stomatal CO2 sensing. Additionally, increasing heat stress functions via a distinct photosynthesis-uncoupled stomatal openingpathway.

The biomechanics of turgor pressure

If you ever forget to water your houseplant, you may find its leaves getting soft and droopy - if you water it again in time, the leaves may stiffen, spring back up, and resist gravity. During this recovery, plant cells absorb water and build up an intracellular pressure, called turgor pressure, similar to inflating a balloon. Turgor pressure is an intrinsic component of plant physiology, and its biomechanical role as the 'hydroskeleton' is generally appreciated either statically in structural stability, like leaves resisting gravity, or dynamically in rapid motions, like Venus flytrap snapping, Mimosa closing, or stomatal opening. Slow but non-static processes like plant cell expansion also rely on turgor pressure, and it has been increasingly realized that turgor pressure and water fluxes in plant tissues play active roles in plant growth and morphogenesis. But where does turgor pressure come from, and how does it interact with other biomechanical properties of a plant cell? This primer aims to answer these questions by taking a brief tour from osmosis, to pressure vessel theory, then plant cell rheology. Although this primer is centered around intracellular pressure in plant cells, the biomechanical concepts are generally applicable to other organisms like bacteria and fungi, and even animal cells, which do not have cell walls, but are either embedded in stiff extracellular matrix (ECM) or are contracting (see the following section). To explain some of these concepts, we included a few equations, most of which are not hard to derive at all. We encourage readers to check the included examples, try for themselves, and look up derivations in suggested further reading materials.

Optimized irrigation practices with fertilizer utilization strategies to improve photo-fluorescence efficiency, vascular bundles and maize production in semi-arid regions

The mechanism of irrigation models combined with fertilizer utilization strategies under the biodegradable film mulching could greatly promote crop photosynthesis, vascular bundles structure; resource utilization and maize production are unclear in semi-arid areas. Unfortunately, this mechanism provides a scientific basis for improving irrigation and fertilizer utilization. A field study was carried out during 2021–2022 years. Seven treatments were established: two nitrogen levels: low-N (150 kg N ha−1) and high-N (300 kg N ha−1) combined with three different irrigation models: drip irrigation (DI), ridge irrigation (RI) and border irrigation (BI) under the biodegradable film mulching with (CK) treatment have no irrigation, fertilizer and mulching. Our results revealed that DIH treatment considerably increased soil water storage, enhanced photosynthesis rate (Pn) of maize by mainly to facilitate stomatal opening compared to the rest of all treatments. In addition, it also enhances the differentiation of the vascular bundle system and maintains its post silk function under better environmental conditions, greatly improving nitrogen storage in soil and plants, and enhancing maize productivity. DIH and RIH treatments significantly increased net photosynthesis rate (Pn), stomatal conductance (Gs), transpiration rate (Tr), maximum quantum yield of PSII photochemistry (Fv/Fm), and effective quantum yield of PSII photochemistry (ΦPSII), photochemical quenching (qP), non-photochemical quenching (NPQ), and yield were observed, but evapotranspiration (ET) decreased at different growth stages. The results showed that DIH treatment was an effective tillage strategy, which increased biomass yield by 32.6 %, grain yield by 46.0 %, water use efficiency (WUE) by 46.2 %, and nitrogen use efficiency (NUE) by 86.4 % compared to other treatments. Given these results, thus we recommend the drip irrigation combined with a high-N level under a biodegradable film mulching increase photo-fluorescence efficiency, maize production and resource utilization efficiency in semi-arid regions.

The PATROL1 function in roots contributes to the increase in shoot biomass

Main conclusionPATOL1 contributes to increasing biomass not only by effective stomatal movement but also by root meristematic activity.PATROL1 (PROTON ATPase TRANSLOCATION CONTROL 1), a protein with a MUN domain, is involved in the intercellular trafficking of AHA1 H+-ATPase to the plasma membrane in guard cells. This allows for larger stomatal opening and more efficient photosynthesis, leading to increased biomass. Although PATROL1 is expressed not only in stomata but also in other tissues of the shoot and root, the role in other tissues than stomata has not been determined yet. Here, we investigated PATROL1 functions in roots using a loss-of-function mutant and an overexpressor. Cytological observations revealed that root meristematic size was significantly smaller in the mutant resulting in the short primary root. Grafting experiments showed that the shoot biomass of the mutant scion was increased when it grafted onto wild-type or overexpressor rootstocks. Conversely, grafting of the overexpressor scion shoot enhanced the growth of the mutant rootstock. The leaf temperatures of the grafted plants were consistent with those of their respective genotypes, indicating cell-autonomous behavior of stomatal movement and independent roles of PATROL1 in plant growth. Moreover, plasma membrane localization of AHA1 was not altered in root epidermal cells in the patrol1 mutant implying existence of a different mode of PATROL1 action in roots. Thus PATROL1 plays a role in root meristem and contributes to increase shoot biomass.

Triacontanol Reverses Abscisic Acid Effects on Stomatal Regulation in Solanum lycopersicum L. under Drought Stress Conditions

When applied under abiotic stress conditions, triacontanol (TRIA) is effective in regulating the physicochemical processes in plants through mechanisms of defence such as abscisic acid (ABA) signalling. However, TRIA’s role in relation to ABA and stomatal opening is unclear. Therefore, the objective of this study was to evaluate the effects of TRIA and ABA and their combinations on different variables related to stomatal regulation in Solanum lycopersicum, which is subjected to drought stress, and on the leaf epidermis. The negative effects of stress and responses triggered by ABA were reversed in plants treated with TRIA. TRIA increased stomatal conductance and photosynthetic activity in the early hours, and it was determined that TRIA produced larger stomata than did the other treatments. Moreover, the chloroplasts of plants treated with TRIA were significantly smaller and more numerous than those of the control, which could improve CO2 diffusion efficiency and may be related to the regulation of stomatal opening and photosynthesis. Finally, the abaxial epidermis tests reaffirmed the inhibitory effects of TRIA on ABA on stomatal opening. These results confirm the important role of TRIA in regulating various processes in plants and processes triggered by ABA, such as those related to stomatal regulation.

The Arabidopsis KASH protein SINE3 is involved in male and female gametogenesis.

The Arabidopsis KASH protein SINE3 is involved in male and female gametophyte development, likely affecting the first post-meiotic mitosis in both cases, and is required for full seed set. Linker of nucleoskeleton and cytoskeleton (LINC) complexes are protein complexes spanning the inner and outer membranes of the nuclear envelope (NE) and are key players in nuclear movement and positioning. Through their roles in nuclear movement and cytoskeletal reorganization, plant LINC complexes affect processes as diverse as pollen tube rupture and stomatal development and function. KASH proteins are the outer nuclear membrane component of the LINC complex, with conserved C-termini but divergent N-terminal cytoplasmic domains. Of the known Arabidopsis KASH proteins, SUN-INTERACTING NUCLEAR ENVELOPE PROTEIN 3 (SINE3) has not been functionally characterized. Here, we show that SINE3 is expressed at all stages of male and female gametophyte development. It is located at the NE in male and female gametophytes. Loss of SINE3 results in a female-derived seed set defect, with sine3 mutant ovules arresting at stage FG1. Pollen viability is also significantly reduced, with microspores arresting prior to pollen mitosis I. In addition, sine3 mutants have a minor male meiosis defect, with some tetrads containing more than four spores. Together, these results demonstrate that the KASH protein SINE3 plays a crucial role in male and female gametophyte development, likely affecting the first post-meiotic nuclear division in both cases.

Endomembrane trafficking driven by microtubule growth regulates stomatal movement in Arabidopsis

Microtubule-based vesicle trafficking usually relies upon kinesin and dynein motors and few reports describe microtubule polymerisation driving directional vesicle trafficking. Here we show that Arabidopsis END BINDING1b (EB1b), a microtubule plus-end binding protein, directly interacts with SYP121, a SNARE protein that mediates the trafficking of the K+ channel KAT1 and its distribution to the plasma membrane (PM) in Arabidopsis guard cells. Knockout of AtEB1b and its homologous proteins results in a modest but significant change in the distribution of KAT1 and SYP121 in guard cells and consequently delays light-induced stomatal opening. Live-cell imaging reveals that a portion of SYP121-associated endomembrane compartments co-localise with AtEB1b at the growing ends of microtubules, trafficking along with the growth of microtubules for targeting to the PM. Our study reveals a mechanism of vesicle trafficking driven by microtubule growth, which is involved in the redistribution of PM proteins to modulate guard cell movement.