Stomatal Closure Research Articles

The stomatal response to vapor pressure deficit drives the apparent temperature response of photosynthesis in tropical forests.

As temperature rises, net carbon uptake in tropical forests decreases, but the underlying mechanisms are not well understood. High temperatures can limit photosynthesis directly, for example by reducing biochemical capacity, or indirectly through rising vapor pressure deficit (VPD) causing stomatal closure. To explore the independent effects of temperature and VPD on photosynthesis we analyzed photosynthesis data from the upper canopies of two tropical forests in Panama with Generalized Additive Models. Stomatal conductance and photosynthesis consistently decreased with increasing VPD, and statistically accounting for VPD increased the optimum temperature of photosynthesis (Topt) of trees from a VPD-confounded apparent Topt of c. 30-31°C to a VPD-independent Topt of c. 33-36°C, while for lianas no VPD-independent Topt was reached within the measured temperature range. Trees and lianas exhibited similar temperature and VPD responses in both forests, despite 1500 mm difference in mean annual rainfall. Over ecologically relevant temperature ranges, photosynthesis in tropical forests is largely limited by indirect effects of warming, through changes in VPD, not by direct warming effects of photosynthetic biochemistry. Failing to account for VPD when determining Topt misattributes the underlying causal mechanism and thereby hinders the advancement of mechanistic understanding of global warming effects on tropical forest carbon dynamics.

Impact of Fruit Load on the Replenishment Dynamics of Internal Water Reserves in Olive Trees

Stem refilling has been studied in many forest species, but its impact on olive trees remains underexplored. This study aims to examine the effect of varying fruit loads on stem refilling rates in olive trees. The experiment was conducted in a commercial olive orchard over two years, characterized by a biennial bearing cycle with minimal fruit production in 2021 (“OFF” year) and maximal fruit production in 2022 (“ON” year). Sap flow sensors measured the water volume differences traversing the apex and base of main branches in two experimental trees. Tree water status was monitored using psychrometers, and soil moisture content was continuously recorded. Results suggest that alternate bearing significantly affects the stem refilling process, while soil moisture availability also plays a pivotal role. During the “ON” year, water reserve consumption increased to 63.6% compared to the “OFF” year to meet the water requirements of developing fruits. Replenishment occurred at various times throughout the 24 h period, including early morning, afternoon, and night when stomatal conductance measurements indicated stomatal closure. During the “ON” year, olive trees experienced intense nocturnal replenishment of reserves, regardless of soil moisture, water vapor pressure deficit, or fruit development stage. These findings offer novel insights into olive trees’ rehydration dynamics that can be used to optimize irrigation scheduling and improve water use efficiency.

The microRNA408–plantacyanin module balances plant growth and drought resistance by regulating reactive oxygen species homeostasis in guard cells

Abstract The conserved microRNA (miRNA) miR408 enhances photosynthesis and compromises stress tolerance in multiple plants, but the cellular mechanism underlying its function remains largely unclear. Here, we show that in Arabidopsis (Arabidopsis thaliana), the transcript encoding the blue copper protein PLANTACYANIN (PCY) is the primary target for miR408 in vegetative tissues. PCY is preferentially expressed in the guard cells, and PCY is associated with the endomembrane surrounding individual chloroplasts. We found that the MIR408 promoter is suppressed by multiple abscisic acid (ABA)-responsive transcription factors, thus allowing PCY to accumulate under stress conditions. Genetic analysis revealed that PCY elevates reactive oxygen species (ROS) levels in the guard cells, promotes stomatal closure, reduces photosynthetic gas exchange, and enhances drought resistance. Moreover, the miR408–PCY module is sufficient to rescue the growth and drought tolerance phenotypes caused by gain- and loss-of-function of MYB44, an established positive regulator of ABA responses, indicating that the miR408–PCY module relays ABA signaling for regulating ROS homeostasis and drought resistance. These results demonstrate that miR408 regulates stomatal movement to balance growth and drought resistance, providing a mechanistic understanding of why miR408 is selected during land plant evolution and insights into the long-pursued quest of breeding drought-tolerant and high-yielding crops.

Genome-wide analysis of the poplar homeobox gene family and the identification of PtHB180 as an important regulator of plant developmental processes and drought stress response

Homeobox (HB) transcription factors have been reported to regulate various developmental processes and hormone signaling and mediate plant adaption to abiotic stresses, including drought. As a major subfamily of HB family, KNOX genes have been widely studied in herbaceous plants. However, the behaviors of KNOX genes in regulating plant developmental processes and stress resistance in woody plants remain poorly understood. Here, we found a KNOX family gene in the model tree species Populus trichocarpa (hereafter “poplar”), PtHB180, which enhances drought tolerance by regulating developmental processes through auxin and gibberellin pathways and mediating the ABA signaling pathway leading to rapid stomatal closure. Overexpression lines of PtHB180 (PtHB180-OE) displayed pleiotropic phenotypes, including increased plant height and decreased leaf area by affecting auxin and/or gibberellin contents and signaling. PtHB180-OE plants also displayed improved drought tolerance under short- and long-term drought stress. Results of transcriptome sequencing suggested the involvement of genes related to ABA signaling in regulating drought tolerance in PtHB180-OE. Specifically, PtHB180-OE plants showed significantly enhanced stomatal sensitivity to exogenous ABA. Taken together, our results indicate that PtHB180 enhances drought tolerance in poplar via regulating plant architecture and mediating ABA signaling to close stomata. Therefore, PtHB180 can be used as a potential candidate gene to improve drought tolerance by genetic transformation in poplar and related plant species.

CO2 Response Screen in Grass Brachypodium Reveals Key Role of a MAP Kinase in CO2-Triggered Stomatal Closure.

Plants respond to increased CO2 concentrations through stomatal closure, which can contribute to increased water use efficiency. Grasses display faster stomatal responses than eudicots due to dumbbell-shaped guard cells flanked by subsidiary cells working in opposition. However, forward genetic screening for stomatal CO2 signal transduction mutants in grasses has yet to be reported. The grass model Brachypodium distachyon is closely related to agronomically important cereal crops, sharing largely collinear genomes. To gain insights into CO2 control mechanisms of stomatal movements in grasses, we developed an unbiased forward genetic screen with an EMS-mutagenized Brachypodium distachyon M5 generation population using infrared imaging to identify plants with altered leaf temperatures at elevated CO2. Among isolated mutants, a "chill1" mutant exhibited cooler leaf temperatures than wildtype Bd21-3 parent control plants after exposure to increased [CO2]. chill1 plants showed strongly impaired high CO2-induced stomatal closure despite retaining a robust abscisic acid-induced stomatal closing response. Through bulked segregant whole-genome-sequencing analyses followed by analyses of further backcrossed F4 generation plants and generation and characterization of sodium-azide and CRISPR-cas9 mutants, chill1 was mapped to a protein kinase, Mitogen-Activated Protein Kinase 5 (BdMPK5). The chill1 mutation impaired BdMPK5 protein-mediated CO2/HCO3- sensing together with the High Temperature 1 (HT1) Raf-like kinase in vitro. Furthermore, AlphaFold2-directed structural modeling predicted that the identified BdMPK5-D90N chill1 mutant residue is located at the interface of BdMPK5 with the BdHT1 Raf-like kinase. BdMPK5 is a key signaling component that mediates CO2-induced stomatal movements and is proposed to function as a component of the primary CO2 sensor in grasses.

H2S is involved in drought-mediated stomatal closure through PLDα1 in Arabidopsis.

PLDα1 promoted H2S production by positively regulating the expression of LCD. Stomatal closure promoted by PLDα1 required the accumulation of H2S under drought stress. Phospholipase Dα1 (PLDα1) acting as one of the signal enzymes can respond to drought stress. It is well known that hydrogen sulfide (H2S) plays an important role in plant responding to biotic or abiotic stress. In this study, the functions and relationship between PLDα1 and H2S in drought stress resistance in Arabidopsis were explored. Our results indicated that drought stress promotes PLDα1 and H2S production by inducing the expression of PLDα1 and LCD genes. PLDα1 and LCD enhanced plant tolerance to drought by regulating membrane lipid peroxidation, proline accumulation, H2O2 content and stomatal closure. Under drought stress, the H2O2 content of PLDα1-deficient mutant (pldα1), L-cysteine desulfhydrase (LCD)-deficient mutant (lcd) was higher than that of ecotype (WT), the stomatal aperture of pldα1 and lcd was larger than that of WT. The transcriptional and translational levels of LCD were lower in pldα1 than that in WT. Exogenous application of the H2S donor NaHS or GYY reduced the stomatal aperture of WT, pldα1, PLDα1-CO, and PLDα1-OE lines, while exogenous application of the H2S scavenger hypotaurine (HT) increased the stomatal aperture. qRT-PCR analysis of stomatal movement-related genes showed that the expression of CAX1, ABCG5, SCAB1, and SLAC1 genes in pldα1 and lcd were down-regulated, while ACA1 and OST1 gene expression was significantly up-regulated. Thus, PLDα1 and LCD are required for stomatal closure to improve drought stress tolerance.

Leaf water relations determine the trade-off between ozone resistance and stomatal functionality in urban tree species.

Urban trees possess different capacities to mitigate ozone (O3) pollution through stomatal uptake. Stomatal closure protects trees from oxidative damage but limits their growth. To date, it is unclear how plant hydraulic function affect stomatal behaviour and determine O3 resistance. We assessed gas exchange and hydraulic traits in three subtropical urban tree species, Celtis sinensis, Quercus acutissima, and Q. nuttallii, under nonfiltered ambient air (NF) and elevated O3 (NF60). NF60 decreased photosynthetic rate (An) and stomatal conductance (gs) only in Q. acutissima and Q. nuttallii. Maintained An in C. sinensis suggested high O3 resistance and was attributed to higher leaf capacitance at the full turgor. However, this species exhibited a reduced stomatal sensitivity to vapour pressure deficit and an increased minimal gs under NF60. Such stomatal dysfunction did not decrease intrinsic water use efficiency (WUE) due to a tight coupling of An and gs. Conversely, Q. acutissima and Q. nuttallii showed maintained stomatal sensitivity and increased WUE, primarily correlated with gs and leaf water relations, including relative water content and osmotic potential at turgor loss point. Our findings highlight a trade-off between O3 resistance and stomatal functionality, with efficient stomatal control reducing the risk of hydraulic failure under combined stresses.

A network-based modeling framework reveals the core signal transduction network underlying high carbon dioxide-induced stomatal closure in guard cells.

Stomata are pores on plant aerial surfaces, each bordered by a pair of guard cells. They control gas exchange vital for plant survival. Understanding how guard cells respond to environmental signals such as atmospheric carbon dioxide (CO2) levels is not only insightful to fundamental biology but also relevant to real-world issues of crop productivity under global climate change. In the past decade, multiple important signaling elements for stomatal closure induced by elevated CO2 have been identified. Yet, there is no comprehensive understanding of high CO2-induced stomatal closure. In this work, we assemble a cellular signaling network underlying high CO2-induced stomatal closure by integrating evidence from a comprehensive literature analysis. We further construct a Boolean dynamic model of the network, which allows in silico simulation of the stomatal closure response to high CO2 in wild-type Arabidopsis thaliana plants and in cases of pharmacological or genetic manipulation of network nodes. Our model has a 91% accuracy in capturing known experimental observations. We perform network-based logical analysis and reveal a feedback core of the network, which dictates cellular decisions in closure response to high CO2. Based on these analyses, we predict and experimentally confirm that applying nitric oxide (NO) induces stomatal closure in ambient CO2 and causes hypersensitivity to elevated CO2. Moreover, we predict a negative regulatory relationship between NO and the protein phosphatase ABI2 and find experimentally that NO inhibits ABI2 phosphatase activity. The experimental validation of these model predictions demonstrates the effectiveness of network-based modeling and highlights the decision-making role of the feedback core of the network in signal transduction. We further explore the model's potential in predicting targets of signaling elements not yet connected to the CO2 network. Our combination of network science, in silico model simulation, and experimental assays demonstrates an effective interdisciplinary approach to understanding system-level biology.

Cultivar-specific regulation of antioxidant enzyme activity and stomatal closure confer tolerance of wheat to elevated ozone: A two-year open-field study with five cultivars

Elevated concentrations of tropospheric ozone (O3) pose a significant threat to food production in many regions of the world. We conducted a two-year experiment with five winter wheat (Triticum aestivum L.) cultivars, Yangmai16 (Y16), Yangmai23 (Y23), Zhenmai12 (Z12), Yangmai29 (Y29), and Yangmai30 (Y30), to assess photosynthetic traits, lipid peroxidation, and antioxidant systems under ambient (A-O3) or elevated (E-O3; 1.5 times A-O3) O3 in a Free-Air O3-Concentration Enrichment system (FACE). E-O3 accelerated leaf senescence, manifested by marked reductions in photosynthetic rate, stomatal conductance, total antioxidant capacity, and photosynthetic pigment content, accompanied by altered antioxidant enzyme activity and a significant increase in malondialdehyde content (an indicator of lipid peroxidation). Most parameters showed significant inter-cultivar differences, as well as interactions between O3 and cultivar. The response of the antioxidant system in wheat flag leaves was influenced by O3 exposure levels. The response of antioxidant enzymes at the grain filling stage and the timely closure of stomata conferred tolerance to O3. This tolerance was associated with higher antioxidant enzyme activity at the anthesis stage but was not related to the size of stomata. This study provides a crucial theoretical foundation for further breeding O3-tolerant cultivars.

Genome-wide identification following functional analysis of amino acid permease and cationic amino acid transporter gene families in maize and their role in drought stress

In maize, the amino acid permease (AAP) and the cationic amino acid transporters (CAT) play a significant role in amino nitrogen (N) uptake, transport, utilization, and homeostasis. Hence, genome-wide analysis of maize AAP and CAT resulted in 24 AAP (1-24) and 14 CAT (1-14) proteins in maize having significant variation in molecular weights. Most proteins are basic and localized in the plasma membrane. AAPs and CATs showed up to 7 and 14 introns respectively. All proteins showed multiple motifs, alpha-helices, transmembrane helices (except ZmAAP23), and coiled in nature. The ZmAAP and ZmCAT promoters consisted of phytohormone, plant growth and development, and stress-related cis-elements. All genes are distributed in different chromosomes. All the above properties are consistent with the same in Arabidopsis, which represent their functional similarities. Significantly higher transcription of some AAPs and CATs in vegetative and/or reproductive tissues reveal their great potential to source-sink transport and reuptake of leaked amino N from the rhizosphere. Interestingly, some members of the above-mentioned families are also upregulated during drought stress revealing their great role in drought tolerance or acclimatization during the stress. For increasing drought tolerance, these transporters might interact with the genes regulating the turgor pressure of the guard cells for increasing stomatal closure and reducing transpiration in leaves through controlling cellular osmotic homeostasis.

Water use and mortality risk of four tropical canopy trees with different leaf phenology during the 2016 El Niño drought

The physiological response of plants to water stresses has been a focus in understanding plant-atmosphere feedback. Tropical forests are particularly noteworthy for their remarkable diversity and dynamics. However, a deep understanding of physiological response and associated mortality risk of tropical trees with different leaf phenology under drought is still deficient. In this study, we combined sap flow measurements and a plant hydraulic model to present the water utilization of four canopy trees (two deciduous trees and two evergreen trees) in a Panamanian seasonal tropical forest during the 2016 El Niño drought. The results showed that the transpiration (Td) of deciduous trees rapidly decreased with intensifying soil drought during the dry season, while evergreen trees still maintained high Td and increased with the increase of atmospheric dryness (vapor pressure deficit (VPD)). During the wet season, the Td of both deciduous and evergreen trees was jointly driven by increasing soil moisture (SWC) and reducing VPD. The differential Td pattern during the dry season is closely related to the difference in leaf phenology, with deciduous trees greatly reducing canopy stomatal conductance (Gs) through defoliation, and thereby maintaining leaf water potential (Ψl) and reducing water loss. Additionally, deciduous trees demonstrated a slow increase in the difference between midday (Ψl,md) and predawn (Ψl,pd) leaf water potential (i.e. lower hydraulic sensitivity (σ)) compared to evergreen trees. Evaluation of mortality risk under changing water stresses further indicated that all trees exhibited high hydraulic failure risk (HFR) under decreasing SWC scenarios. Moreover, deciduous trees displayed a much higher stomatal closure risk (SCR) than evergreen trees. These differences in Gs response and σ suggest a key role of the coordination between stomatal regulation and plant hydraulics in mediating physiological response to water stresses. More investigation and mechanism illumination will facilitate the prediction of tropical forest response to drought.

Lethal combination for seedlings: extreme heat drives mortality of drought-exposed high-elevation pine seedlings.

Hotter drought- and biotically-driven tree mortality are expected to increase with climate change in much of the western United States, and species persistence will depend upon ongoing establishment under novel conditions or migration to track ecological niche requirements. High-elevation tree species may be particularly vulnerable to increasing water stress as snowpack declines, increasing the potential for adult mortality and simultaneous regeneration failures. Seedling survival will be determined by ecophysiological limitations in response to changing water availability and temperature. We exposed seedlings from populations of Pinus longaeva, Pinus flexilis, and Pinus albicaulis to severe drought and concurrent temperature stress in common gardens testing timing of drought onset under two different temperature regimes. We monitored seedling functional traits, physiological function, and survival. The combined stressors of water limitation and extreme heat led to conservative water use strategies and declines in physiological function, with these joint stressors ultimately exceeding species' tolerances and leading to complete episodic mortality across all species. Growing conditions were the primary determinant of seedling trait expression, with seedlings exhibiting more drought-resistant traits such as lower specific leaf area in the hottest, driest treatment conditions. Water stress-induced stomatal closure was also widely apparent. Under adequate soil moisture, seedlings endured prolonged exposure to high air and surface temperatures, suggesting broad margins for survival. The critical interaction between soil moisture and temperature suggests that rising temperatures will exacerbate growing season moisture stress. Our results highlight the importance of local conditions over population- and species-level influences in shaping strategies for stress tolerance and resistance to desiccation at this early life stage. By quantifying some of the physiological consequences of drought and heat that lead to seedling mortality, we can better understand the future effects of global change on the composition and distribution of high-elevation conifer forests.

Spider mite herbivory induces an abscisic acid-driven stomatal defense.

Arthropod herbivory poses a serious threat to crop yield, prompting plants to employ intricate defense mechanisms against pest feeding. The generalist pest two-spotted spider mite (Tetranychus urticae) inflicts rapid damage and remains challenging due to its broad target range. In this study, we explored the Arabidopsis (Arabidopsis thaliana) response to T. urticae infestation, revealing the induction of abscisic acid (ABA), a hormone typically associated with abiotic stress adaptation, and stomatal closure during water stress. Leveraging a FRET-based ABA biosensor (nlsABACUS2-400n), we observed elevated ABA levels in various leaf cell types post-mite feeding. While ABA's role in pest resistance or susceptibility has been debated, an ABA-deficient mutant exhibited increased mite infestation alongside intact canonical biotic stress signaling, indicating an independent function of ABA in mite defense. We established that ABA-triggered stomatal closure effectively hinders mite feeding and minimizes leaf cell damage through genetic and pharmacological interventions targeting ABA levels, ABA signaling, stomatal aperture, and density. This study underscores the critical interplay between biotic and abiotic stresses in plants, highlighting how the vulnerability to mite infestation arising from open stomata, crucial for transpiration and photosynthesis, reinforces the intricate relationship between these stress types.

Alternative splicing as a driver of natural variation in abscisic acid response.

Abscisic acid (ABA) is a crucial player in plant responses to the environment. It accumulates under stress, activating downstream signaling to implement molecular responses that restore homeostasis. Natural variance in ABA sensitivity remains barely understood, and the ABA pathway has been mainly studied at the transcriptional level, despite evidence that posttranscriptional regulation, namely, via alternative splicing, contributes to plant stress tolerance. Here, we identified the Arabidopsis accession Kn-0 as less sensitive to ABA than the reference Col-0, as shown by reduced effects of the hormone on seedling establishment, root branching, and stomatal closure, as well as by decreased induction of ABA marker genes. An in-depth comparative transcriptome analysis of the ABA response in the two variants revealed lower expression changes and fewer genes affected for the least ABA-sensitive ecotype. Notably, Kn-0 exhibited reduced levels of the ABA-signaling SnRK2 protein kinases and lower basal expression of ABA-reactivation genes, consistent with our finding that Kn-0 contains less endogenous ABA than Col-0. ABA also markedly affected alternative splicing, primarily intron retention, with Kn-0 being less responsive regarding both the number and magnitude of alternative splicing events, particularly exon skipping. We find that alternative splicing introduces a more ecotype-specific layer of ABA regulation and identify ABA-responsive splicing changes in key ABA pathway regulators that provide a functional and mechanistic link to the differential sensitivity of the two ecotypes. Our results offer new insight into the natural variation of ABA responses and corroborate a key role for alternative splicing in implementing ABA-mediated stress responses.

Water status dynamics and drought tolerance of juvenile European beech, Douglas fir and Norway spruce trees as dependent on neighborhood and nitrogen supply.

To increase the resilience of forests to drought and other hazards, foresters are increasingly planting mixed stands. This requires knowledge about the drought response of tree species in pure- and mixed-culture neighborhoods. In addition, drought frequently interacts with continued atmospheric nitrogen (N) deposition. To disentangle these factors for European beech, Norway spruce and Douglas fir, we conducted a replicated three-factorial sapling growth experiment with three moisture levels, (high, medium and low), two N levels (high and ambient) and pure and mixed-culture neighborhoods. We measured biomass, stomatal conductance (GS), shoot water potential (at predawn: ΨPD, midday, and turgor loss point: ΨTLP), branch xylem embolism resistance (Ψ50), and minimum epidermal conductance (Gmin). The three species differed most with respect to Gmin (10-fold higher in beech than in the conifers), hydroscape area (larger in beech), and the time elapsed to reach stomatal closure (TΨGS90) and ΨTLP (TTLP; shorter in beech), while Ψ50 and ΨTLP were remarkably similar. Neighborhood (pure vs. mixed-culture) influenced biomass production, water status and hydraulic traits, notably GS (higher in Douglas fir, but lower in spruce and beech, in mixtures than pure culture), hydraulic safety margin (smaller for beech in mixtures), and TΨGS90 and TTLP (shorter for spruce in mixture). High N generally increased GS, but no consistent N effects on leaf water status and hydraulic traits were detected, suggesting that neighbor identity had a larger effect on plant water relations than N availability. We conclude that both tree neighborhood and N availability modulate the drought response of beech, spruce and Douglas fir. Species mixing can alleviate the drought stress of some species, but often by disadvantaging other species. Thus, our study suggests stabilizing and building resilience of production forests against a drier and warmer climate may depend primarily on the right species choice; species mixing can support the agenda.

Decreased irrigation volume, not irrigation placement, promotes accumulation of multiple hormones in cotton leaves during partial rootzone drying

While ABA is often assumed to mediate partial stomatal closure as the soil dries, other plant hormones and hydraulic signals may also be involved. We tested whether irrigation volume (% of crop evapotranspiration, ET) and placement (partial rootzone drying [PRD] or deficit irrigation [DI], which irrigate part or all of the rootzone respectively) affect this signalling by measuring stomatal conductance (gs), leaf and shoot water potential (Ψleaf, Ψshoot), shoot xylem sap ABA concentration ([X-ABA]shoot) and various foliar hormones (ABA, IAA, SA, JA, JA-Ile and cis-OPDA) in cotton plants exposed to different irrigation volumes (100 %ET or 50 %ET) and placements (DI or PRD). Partial rootzone drying caused stomatal closure coincident with sustained foliar ABA accumulation and minimal changes in Ψshoot, but continued soil drying of the dry compartment reversed partial stomatal closure (with gs of 100 %ET PRD plants sometimes greater than well-watered plants). With 100 %ET PRD, partial stomatal closure correlated with decreased soil moisture of the dry compartment and increased [ABA]leaf, but neither Ψleaf nor [X-ABA]shoot. Irrespective of irrigation placement, 50 %ET significantly decreased gs, Ψleaf and Ψshoot, but significantly increased [ABA]leaf, [X-ABA]shoot, [SA]leaf, [IAA]leaf and [cis-OPDA]leaf, with stomatal closure of 50 %ET PRD plants occurring earlier than 50 %ET DI plants. While stomatal closure at 50 %ET correlated with foliar accumulation of multiple plant hormones, foliar ABA dynamics best explained transient stomatal closure at 100 %ET PRD but not stomatal re-opening with prolonged soil drying. Thus, stomatal sensitivity to drying soil (and putative regulatory signals such as ABA) depended on irrigation volume and placement.

Acclimation of key photosynthetic traits drives edible biomass increases in crop species grown in aquaponics

Sustainable agriculture practices may contribute to increasing food demand while alleviating issues with fertilizer over-use and water availability in traditional agriculture. Aquaponics systems provide plants with a constant supply of nutrient dense water from fish efflux. Root uptake filters the fish efflux and water is recycled back to fish tanks, reducing the water needed to produce both plants and fish. Plants acclimate their physiology and anatomy to changing environments, yet, the degree of acclimation in aquaponics is unknown. We tested if acclimation of leaf physiology and anatomy to constant water and nutrient supply in aquaponics affected edible yield. We measured acclimation of leaf gas exchange (Asat, gsw), photosynthetic capacity (Jmax,Vcmax), stomatal density and chlorophyll production by comparing plants grown in aquaponics or soil for three species: Lactuca sativa (Romaine lettuce), Triticum aestivum (Winter Wheat), and Ocimum basilicum (Basil). We hypothesized that non-limiting nitrogen supply in aquaponics would drive increased production of chlorophyll, increasing electron transport rate (Jmax) and carboxylation rate (Vcmax). Additionally, constant water supply in aquaponics would remove limitations driving stomatal closure, leading to higher photosynthesis rates. We found aquaponics plants produced more chlorophyll, driving increased photosynthetic capacity through higher Jmax and Vcmax. Additionally, aquaponics plants increased photosynthesis rates (Asat) via higher stomatal conductance (gsw) of fewer stomata. Acclimation of key photosynthetic traits in aquaponics plants resulted in more edible biomass compared to soil grown plants, often with reduced investment in root production. Overall, aquaponics appears well suited to producing higher yields of certain crop species, while also reducing environmental impacts.

Transcriptomic and metabolomic insights into drought response strategies of two Astragalus species

Drought is a major environmental constraint to plant productivity, and plants cope with water limitation using drought avoidance and/or tolerance strategies. Mesoxerophyte Astragalus mongholicus displayed better drought resistance than the mesophyte A. membranaceus. However, the underlying response mechanisms are still elusive. In this study, we characterized the morphological, physiological, molecular, and metabolic responses to drought stress to reveal the different response strategies in the two Astragalus species. The susceptible A. membranaceus, with a larger leaf area, showed significant stomatal closure at moderate drought stress, indicating drought avoidance strategy to minimize transpiration. A. mongholicus, with a smaller leaf area, was better in maintaining photosynthesis and activating antioxidant enzymes than A. membranaceus towards the drought tolerance strategy. Drought has greater disturbance on gene expression in A. membranaceus, compared to A. mongholicus, indicating that A. membranaceus was more sensitive in the face of drought. Meanwhile, highly conserved osmotic stress signal transduction pathways were discovered in two Astragalus species under drought treatment. The up-regulation of dehydration-responsive element-binding gene (DREB) expression in A. mongholicus, initiating a series of stress response pathways to help plants adapt and cope with water limitation, showed better “tolerance” than in A. membranaceus. An increase of compatible solutes in A. membranaceus indicates a positive response in regulating osmotic homeostasis in response to drought. The greater investment in antioxidant supplementation phenolic compounds of A. mongholicus suggests the “Tolerance” strategy. Furthermore, the greater antioxidant investment of A. mongholicus is evidenced by higher antioxidant efficacies in terms of antiradical activity. In summary, we conclude that the “Tolerance” strategy of A. mongholicus and the “Avoidance” strategy of A. membranaceus in the face of drought stress can be established.

Coordinated hydraulic traits influence the two phases of time to hydraulic failure in five temperate tree species differing in stomatal stringency.

Worldwide, forests are increasingly exposed to extreme droughts causing tree mortality. Because of the complex nature of the mechanisms involved, various traits have been linked to tree drought responses with contrasting results. This may be due to species-specific strategies in regulating water potential, a process that unfolds in two distinct phases: a first phase until stomatal closure, and a second phase until reaching lethal xylem hydraulic thresholds. We conducted dry-down experiments with five broadleaved temperate tree species differing in their degree of isohydry to estimate the time to stomatal closure (tsc) and subsequent time to critical hydraulic failure (tcrit). We measured various traits linked to tree drought responses, such as the water potentials at turgor loss point (Ptlp), stomatal closure (Pgs90) and 12, 50 and 88% loss of xylem hydraulic conductance (P12, P50, P88), hydraulic capacitance (C), minimum leaf conductance (gmin), hydroscape area (HSA) and hydraulic safety margins (HSM). We found that Pgs90 followed previously recorded patterns of isohydry and was associated with HSA. Species ranked from more to less isohydric in the sequence Acer pseudoplatanus<Betula pendula<Tilia cordata<Sorbus aucuparia<Fagus sylvatica. Their degree of isohydry was associated with leaf safety (Ptlp and gmin), drought avoidance (C) and tsc, but decoupled from xylem safety (HSM and P88) and tcrit. Regardless of their stomatal stringency, species with wider HSM and lower P88 reached critical hydraulic failure later. We conclude that the duration of the first phase is determined by stomatal regulation, while the duration of the second phase is associated with xylem safety. Isohydry is thus linked to water use rather than to drought survival strategies, confirming the proposed use of HSA as a complement to HSM for describing plant drought responses before and after stomatal closure.

Phosphatidic acid produced by phospholipase Dα1 and Dδ is incorporated into the internal membranes but not involved in the gene expression of RD29A in the abscisic acid signaling network in Arabidopsis thaliana.

Core protein components of the abscisic acid (ABA) signaling network, pyrabactin resistance (PYR), protein phosphatases 2C (PP2C), and SNF1-related protein kinase 2 (SnRK2) are involved in the regulation of stomatal closure and gene expression downstream responses in Arabidopsis thaliana. Phosphatidic acid (PA) produced by the phospholipases Dα1 and Dδ (PLDs) in the plasma membrane has been identified as a necessary molecule in ABA-inducible stomatal closure. On the other hand, the involvement of PA in ABA-inducible gene expression has been suggested but remains a question. In this study, the involvement of PA in the ABA-inducible gene expression was examined in the model plant Arabidopsis thaliana and the canonical RD29A ABA-inducible gene that possesses a single ABA-responsive element (ABRE) in the promoter. The promoter activity and accumulation of the RD29A mRNA during ABA exposure to the plants were analyzed under conditions in which the production of PA by PLDs is abrogated through chemical and genetic modification. Changes in the subcellular localization of PA during the signal transduction were analyzed with confocal microscopy. The results obtained in this study suggest that inhibition of PA production by the PLDs does not affect the promoter activity of RD29A. PA produced by the PLDs and exogenously added PA in the plasma membrane are effectively incorporated into internal membranes to transduce the signal. However, exogenously added PA induces stomatal closure but not RD29A expression. This is because PA produced by the PLDs most likely inhibits the activity of not all but only the selected PP2C family members, the negative regulators of the RD29A promoter. This finding underscores the necessity for experimental verifications to adapt previous knowledge into a signaling network model before its construction.