Significance of phosphorus deficiency for the mitigation of mercury toxicity in the Robinia pseudoacacia L.- rhizobia symbiotic association.

Nitrogen (N2)-fixing legumes can be used for phytoremediation of toxic heavy metal Mercury (Hg) contaminated soil, but N2-fixation highly relies on phosphorus (P) availability for nodule formation and functioning. Here, we characterized the significance of P deficiency for Hg accumulation and toxicity in woody legume plants. Consequences for foliar and root traits of rhizobia inoculation, Hg exposure (+Hg) and low P (-P) supply, individually and in combination were characterized at both the metabolite and transcriptome levels in seedlings of two Robinia pseudoacacia L. provenances originating from contrasting climate and soil backgrounds, i.e., GS in northwest and the DB in northeast China. Our results reveal that depleted P mitigates the toxicity of Hg at the transcriptional level. In leaves of Robinia depleted P reduced oxidative stress and improved the utilization strategy of C, N and P nutrition; in roots depleted P regulated the expression of genes scavenging oxidative stress and promoting cell membrane synthesis. Rhizobia inoculation significantly improved the performance of both Robinia provenances under individual and combined +Hg and -P by promoting photosynthesis, increasing foliar N and P content and reducing H2O2 and MDA accumulation despite enhanced Hg uptake. DB plants developed more nodules, had higher biomass and accumulated higher Hg amounts than GS plants and thus are suggested as the high potential Robinia provenance for future phytoremediation of Hg contaminated soils with P deficiency.

Integrative multi-omics analyses of date palm (Phoenix dactylifera) roots and leaves reveal how the halophyte land plant copes with sea water.

Date palm (Phoenix dactylifera L.) is able to grow and complete its life cycle while being rooted in highly saline soils. Which of the many well-known salt-tolerance strategies are combined to fine-tune this remarkable resilience is unknown. The precise location, whether in the shoot or the root, where these strategies are employed remains uncertain, leaving us unaware of how the various known salt-tolerance mechanisms are integrated to fine-tune this remarkable resilience. To address this shortcoming, we exposed date palm to a salt stress dose equivalent to seawater for up to 4 weeks and applied integrative multi-omics analyses followed by targeted metabolomics, hormone, and ion analyses. Integration of proteomic into transcriptomic data allowed a view beyond simple correlation, revealing a remarkably high degree of convergence between gene expression and protein abundance. This sheds a clear light on the acclimatization mechanisms employed, which depend on reprogramming of protein biosynthesis. For growth in highly saline habitats, date palm effectively combines various salt-tolerance mechanisms found in both halophytes and glycophytes: "avoidance" by efficient sodium and chloride exclusion at the roots, and "acclimation" by osmotic adjustment, reactive oxygen species scavenging in leaves, and remodeling of the ribosome-associated proteome in salt-exposed root cells. Combined efficiently as in P. dactylifera L., these sets of mechanisms seem to explain the palm's excellent salt stress tolerance.

Antagonistic effect of mercury and excess nitrogen exposure reveals provenance-specific phytoremediation potential of black locust-rhizobia symbiosis.

Interaction of different environmental constrains pose severe threats to plants that cannot be predicted from individual stress exposure. In this context, mercury (Hg), as a typical toxic and hazardous heavy metal, has recently attracted particular attention. Nitrogen (N2)-fixing legumes can be used for phytoremediation of Hg accumulation, whereas N availability could greatly affect its N2-fixation efficiency. However, information on the physiological responses to combined Hg exposure and excess N supply of woody legume species is still lacking. Here, we investigated the interactive effects of rhizobia inoculation, Hg exposure (+Hg), and high N (+N) supply, individually and in combination (+N*Hg), on photosynthesis and biochemical traits in Robinia pseudoacacia L. seedlings of two provenances, one from Northeast (DB) and one from Northwest (GS) China. Our results showed antagonistic effects of combined + N*Hg exposure compared to the individual treatments that were provenance-specific. Compared to individual Hg exposure, combined + N*Hg stress significantly increased foliar photosynthesis (+50.6%) of inoculated DB seedlings and resulted in more negative (-137.4%) δ15N abundance in the roots. Furthermore, combined + N*Hg stress showed 47.7% increase in amino acid N content, 39.4% increase in NR activity, and 14.8% decrease in MDA content in roots of inoculated GS seedlings. Inoculation with rhizobia significantly promoted Hg uptake in both provenances, reduced MDA contents of leaves and roots, enhanced photosynthesis and maintained the nutrient balance of Robinia. Among the two Robinia provenances investigated, DB seedlings formed more nodules, had higher biomass and Hg accumulation than GS seedlings. For example, total Hg concentrations in leaves and roots and total biomass of inoculated DB seedlings were 1.3,1.9 and 3.4 times higher than in inoculated GS seedlings under combined + N*Hg stress, respectively. Therefore, the DB provenance is considered to possess a higher potential for phytoremediation of Hg contamination compared to the GS provenance in environments subjected to N deposition.

Managing nitrogen for sustainable crop production with reduced hydrological nitrogen losses under a winter wheat-summer maize rotation system: an eight-season field study.

Excessive nitrogen (N) application in wheat-maize cropping systems was adjusted towards more sustainable practices to reduce hydrological N losses while maintaining crop yield. In comprehensive quantification of N management effects on crop yield, N use efficiency (NUE), hydrological N losses, and soil nitrate residual across eight seasons, we have added to growing evidence of strategies beneficial for sustainable crop production with lower hydrological N losses. The results show that adjusted N practices enhanced crop yield and NUE, as compared to farmer's practices, but benefits varied with N rates and types. Optimized N treatment (OPT, 180 kg N ha-1 in both maize and wheat seasons) with or without straw returning produced the most crop yield. They increased maize yield by 5.5% and 7.3% and wheat yield by 6.2% and 3.2% on average, as compared to farmer's practice with huge N application (FP, 345 kg N ha-1 and 240 kg N ha-1 in maize and wheat). Regulation of N release through amendment with controlled release urea at a rate of 144 kg N ha-1 crop-1 (CRU treatment) obtained 4.4% greater maize yield than FP, and sustained a similar wheat yield with less N input, resulting in the highest crop NUE. Additionally, CRU was most effective in mitigating hydrological N loss, with 39.5% and 45.5% less leachate N and 31.9% and 35.9% less runoff N loss than FP in maize and wheat seasons. Synthetic N input correlated significantly and positively with runoff and leachate N losses, indicating it was one of the dominant factors driving hydrological N losses. Moreover, compared to OPT, additional straw returning (STR) or substituting 20% of the nutrients by duck manure (DMS) further reduced runoff N discharges due to the fact that organic matter incorporation increased resilience to rainfall. N over-application in FP caused considerable nitrate accumulation in the 0-90-cm soil profile, while the adjusted N practices, i.e., OPT, STR, CRU, and DMS treatments effectively controlled it to a range of 79.6-92.9 kg N ha-1. This study suggests that efforts using optimized N treatment integrated with CRU or straw returning should be encouraged for sustainable crop production in this region.

Double gains: Boosting crop productivity and reducing carbon footprints through maize-legume intercropping in the Yellow River Delta, China.

The increasing demand for environmentally friendly agricultural practices has driven the need for diversified crop cultivation to optimize crop productivity while minimizing carbon footprints (CFs). However, the impacts of crop diversification on crop production and environmental benefits are still poorly understood. In this study, conducted at two sites in the Yellow River Delta, China, we investigated the effects of legume intercropping, specifically maize/soybean (M/S) and maize/peanut (M/P) systems, on crop productivity, economic return, ecosystem economic budget (NEEB), CF, and carbon sustainability index (CSI) in comparison to conventional monocrops. Crops were grown in replicated field plots and fertilized in their strips according to common practice for monocrops. Compared to the expected averages of monocrops, maize/legume intercropping demonstrated higher crop yields, with M/S achieving a 37% and 43% increase at the two sites, respectively, and M/P achieving an 11% and 20% increase. The higher overyielding in M/S was attributed to stronger selection effects, i.e., interspecific facilitation. However, the complementarity effects induced by the competitive dominance of maize were similar in both intercropping systems. Additionally, M/S exhibited greater potential for improving net revenues compared to M/P. Life cycle assessments revealed lower CFs in the intercropping systems compared to monocultures. M/S reduced CFs per unit of area by 26.8% at both sites, CFs per unit of maize equivalent energy yield by 25% and 33%, and CFs per unit of revenue by 20% and 25% at the two sites, respectively. M/P also resulted in reduced CFs, albeit to a lesser extent. Intercropping enhanced the CSI, with the highest values observed in the M/S system. However, both intercropping systems showed limited effects on soil C sequestration. Overall, our results highlight that maize/legume intercropping is a feasible approach to enhance crop productivity while reducing CFs. The M/S system outperformed the M/P system in terms of crop yields, economic benefits, and CF reduction. However, the intercropping systems showed limited effects on SOC storage. This study provides important implications for sustainable agriculture by appropriate crop diversification.

Plants 'cry' for help through acoustic signals.

Plants perceive sounds, while responses to these sounds were already known. A breakthrough is the discovery by Khait et al. that stressed plants emit various informative ultrasonic sound signals, which can be categorized according to plant species, stress type, and stress severity. This discovery may change how plants are cultivated.

Lipid remodeling of contrasting maize (Zea mays L.) hybrids under repeated drought.

The role of recovery after drought has been proposed to play a more prominent role during the whole drought-adaption process than previously thought. Two maize hybrids with comparable growth but contrasting physiological responses were investigated using physiological, metabolic, and lipidomic tools to understand the plants' strategies of lipid remodeling in response to repeated drought stimuli. Profound differences in adaptation between hybrids were discovered during the recovery phase, which likely gave rise to different degrees of lipid adaptability to the subsequent drought event. These differences in adaptability are visible in galactolipid metabolism and fatty acid saturation patterns during recovery and may lead to a membrane dysregulation in the sensitive maize hybrid. Moreover, the more drought-tolerant hybrid displays more changes of metabolite and lipid abundance with a higher number of differences within individual lipids, despite a lower physiological response, while the responses in the sensitive hybrid are higher in magnitude but lower in significance on the level of individual lipids and metabolites. This study suggests that lipid remodeling during recovery plays a key role in the drought response of plants.

Feeding the world sustainably: efficient nitrogen use.

Globally, overuse of nitrogen (N) fertilizers in croplands is causing severe environmental pollution. In this context, Gu et al. suggest environmentally friendly and cost-effective N management practices and Hamani et al. highlight the use of microbial inoculants to improve crop yields, while reducing N-associated environmental pollution and N-fertilizer use.

Why do plants blush when they are hungry?

Foliar anthocyanins, as well as other secondary metabolites, accumulate transiently under nutritional stress. A misconception that only nitrogen or phosphorus deficiency induces leaf purpling/reddening has led to overuse of fertilizers that burden the environment. Here, we emphasize that several other nutritional imbalances induce anthocyanin accumulation, and nutrient-specific differences in this response have been reported for some deficiencies. A range of ecophysiological functions have been attributed to anthocyanins. We discuss the proposed functions and signalling pathways that elicit anthocyanin synthesis in nutrient-stressed leaves. Knowledge from the fields of genetics, molecular biology, ecophysiology and plant nutrition is combined to deduce how and why anthocyanins accumulate under nutritional stress. Future research to fully understand the mechanisms and nuances of foliar anthocyanin accumulation in nutrient-stressed crops could be utilized to allow these leaf pigments to act as bioindicators for demand-oriented application of fertilizers. This would benefit the environment, being timely due to the increasing impact of the climate crisis on crop performance.

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.

Alkali salt stress causes fast leaf apoplastic alkalinization together with shifts in ion and metabolite composition and transcription of key genes during the early adaptive response of Vicia faba L.

The mechanisms by which plants respond to alkali salt stress are still obscure, and the relevance of alkaline pH under combined alkali salt stress. Early stress responses can indicate mechanisms leading to damage and plant resistance. The apoplast contains essential determinants for plant growth, specifically early apoplastic pH fluctuations are induced by many stressors and hypothesized to be involved in stress signalling. Hence, this study aims to identify fast responses specific to alkaline pH and alkali salt stress by exposing the root of hydroponically grown Vicia faba L. plants to 150 min of either 50 mM NaHCO3 (pH 9) treatment or alkaline pH 9 alone. Apoplastic pH was monitored in real-time by ratiometric fluorescence microscopy simultaneously with SWIR transmission-based measurements of leaf water content (LWC). Moreover, we examined the effect of these stresses on apoplastic, symplastic and xylem ion and metabolite composition together with transcriptions of certain stress-responsive genes. Physiological and transcriptional changes were observed in response to NaHCO3 but not to alkaline pH alone. NaHCO3 elicited a transient reduction in LWC, followed by a transient alkalinization of the apoplast and stomatal closure. Simultaneously, organic acids and sugars accumulated. Fast upregulation of stress-responsive genes showed the significance of gene regulation for early plant adaptation to alkali salt stress.

Under salt stress guard cells rewire ion transport and abscisic acid signaling.

Soil salinity is an increasingly global problem which hampers plant growth and crop yield. Plant productivity depends on optimal water-use efficiency and photosynthetic capacity balanced by stomatal conductance. Whether and how stomatal behavior contributes to salt sensitivity or tolerance is currently unknown. This work identifies guard cell-specific signaling networks exerted by a salt-sensitive and salt-tolerant plant under ionic and osmotic stress conditions accompanied by increasing NaCl loads. We challenged soil-grown Arabidopsis thaliana and Thellungiella salsuginea plants with short- and long-term salinity stress and monitored genome-wide gene expression and signals of guard cells that determine their function. Arabidopsis plants suffered from both salt regimes and showed reduced stomatal conductance while Thellungiella displayed no obvious stress symptoms. The salt-dependent gene expression changes of guard cells supported the ability of the halophyte to maintain high potassium to sodium ratios and to attenuate the abscisic acid (ABA) signaling pathway which the glycophyte kept activated despite fading ABA concentrations. Our study shows that salinity stress and even the different tolerances are manifested on a single cell level. Halophytic guard cells are less sensitive than glycophytic guard cells, providing opportunities to manipulate stomatal behavior and improve plant productivity.

Phenolic compound abundance in Pak choi leaves is controlled by salinity and dependent on pH of the leaf apoplast.

Onset of salinity induces the pH of the leaf apoplast of Pak choi transiently to increase over a period of 2 to 3 hr. This pH event causes protein abundances in leaves to increase. Among them are enzymes that are key for the phenylpropanoid pathway. To answer the questions whether this short-term salt stress also influences contents of the underlying phenylpropanoids and for clarifying as to whether the apoplastic pH transient plays a role for such a putative effect, Pak choi plants were treated with 37.5 mM CaCl2 against a non-stressed control. A third experimental group, where the leaf apoplast of plants treated with 37.5 mM CaCl2, was clamped in the acidic range by means of infiltration of 5 mM citric acid/sodium citrate (pH 3.6), enabled validation of pH-dependent effects. Microscopy-based live cell imaging was used to quantify leaf apoplastic pH in planta. Phenolics were quantified shortly after the formation of the leaf apoplastic pH transient by means of HPLC-DAD-ESI-MS. Results showed that different phenolic compounds were modulated at 150 and 200 min after the onset of chloride salinity. A pH-independent reduction in phenolic acid abundance as well as an accumulation of phenolic acid:malate conjugates was quantified after 200 min of salt stress. However, at 150 min after the onset of salt stress, flavonoids were significantly reduced by salinity in a pH-dependent manner. These results provided a strong indication that the pH of the apoplast is a relevant component for the short-term metabolic response to chloride salinity.

Acclimatisation of guard cell metabolism to long-term salinity.

Stomatal movements are enabled by changes in guard cell turgor facilitated via transient accumulation of inorganic and organic ions imported from the apoplast or biosynthesized within guard cells. Under salinity, excess salt ions accumulate within plant tissues resulting in osmotic and ionic stress. To elucidate whether (a) Na+ and Cl- concentrations increase in guard cells in response to long-term NaCl exposure and how (b) guard cell metabolism acclimates to the anticipated stress, we profiled the ions and primary metabolites of leaves, the apoplast and isolated guard cells at darkness and during light, that is, closed and fully opened stomata. In contrast to leaves, the primary metabolism of guard cell preparations remained predominantly unaffected by increased salt ion concentrations. Orchestrated reductions of stomatal aperture and guard cell osmolyte synthesis were found, but unlike in leaves, no increases of stress responsive metabolites or compatible solutes occurred. Diverging regulation of guard cell metabolism might be a prerequisite to facilitate the constant adjustment of turgor that affects aperture. Moreover, the photoperiod-dependent sucrose accumulation in the apoplast and guard cells changed to a permanently replete condition under NaCl, indicating that stress-related photosynthate accumulation in leaves contributes to the permanent closing response of stomata under stress.

One-time abscisic acid priming induces long-term salinity resistance in Vicia faba: Changes in key transcripts, metabolites, and ionic relations.

Abscisic acid (ABA) priming is known to enhance plant growth and survival under salinity. However, the mechanisms mediating this long-term acclimatization to salt stress are still obscure. Specifically, the long-term transcriptional changes and their effects on ion relations were never investigated. This motivated us to study the long-term (8 days) effect of one-time 24 h root priming treatment with 10 µM ABA on transcription levels of relevant regulated key genes, osmotically relevant metabolites, and ionic concentrations in Vicia faba grown under 50 mM NaCl salinity. The novelty of this study is that we could demonstrate long-term effects of a one-time ABA application. ABA-priming was found to prevent the salt-induced decline in root and shoot dry matter, improved photosynthesis, and inhibited terminal wilting of plants. It substantially increased the mRNA level of AAPK and 14-3-3 ABA inducible kinases and ion transporters (PM H+ -ATPase, VFK1, KUP7, SOS1, and CLC1). These ABA-induced transcriptional changes went along with altered tissue ion patterns. Primed plants accumulated less Na+ and Cl- but more K+ , Ca2+ , Zn2+ , Fe2+ , Mn2+ , NO3 - , and SO4 2- . Priming changed the composition pattern of organic osmolytes under salinity, with glucose and fructose being dominant in unprimed, whereas sucrose was dominant in the primed plants. We conclude that one-time ABA priming mitigates salt stress in Vicia faba by persistently changing transcription patterns of key genes, stabilizing the ionic and osmotic balance, and improving photosynthesis and growth.

Leaf apoplastic alkalization promotes transcription of the ABA-synthesizing enzyme Vp14 and stomatal closure in Zea mays.

The chloride component of NaCl salinity causes the leaf apoplast to transiently alkalinize. This transition in pH reduces stomatal aperture. However, whether this apoplastic pH (pHapo) transient initiates stomatal closure by interacting with other chloride stress-induced responses or whether the pH transient alone initiates stomatal closure is unknown. To clarify the problem, the transient alkalinization of the leaf apoplast was mimicked in intact maize (Zea mays L.) by infiltrating near-neutral pH buffers into the leaf apoplast. Effects of the pHapo transient could thus be investigated independently from other chloride stress-derived effects. Microscopy-based ratiometric live pHapo imaging was used to monitor pHapoin planta. LC-MS/MS and real-time quantitative reverse transcription-PCR leaf analyses showed that the artificially induced pHapo transient led to an increase in the concentrations of the stomata-regulating plant hormone abscisic acid (ABA) and in transcripts of the key ABA-synthesizing gene ZmVp14 in the leaf. Since stomatal aperture and stomatal conductance decreased according to pHapo, we conclude that the pHapo transient alone initiates stomatal closure. Therefore, the functionality does not depend on interactions with other compounds induced by chloride stress. Overall, our data indicate that the pH of the leaf apoplast links chloride salinity with the control of stomatal aperture via effects exerted on the transcription of ABA.

The root as a sink for chloride under chloride-salinity.

Maize has to avoid excess tissue accumulation of Cl- to withstand conditions of Cl--salinity. Restriction of loading of Cl- into the root xylem is one mechanism to keep shoot Cl--concentrations low. The proportion of Cl- that reaches the shoot has to be stored away from the primary site of photosynthesis and growth. We tested whether or not maize is able to re-translocate significant amounts of Cl- from shoot back to root and out into the rooting media. Ion analysis revealed that maize cannot re-translocate Cl-; however, it is stored in sheaths of the old leaves and, surprisingly, in roots. Sequestration of Cl- in the roots might be a strategy to keep concentrations low in young growing shoot tissues and in leaf blades where photosynthesis is running.

Early Changes in Nitrate Uptake and Assimilation Under Drought in Relation to Transpiration.

Soil drying combined with nitrogen (N) deficiency poses a grave threat to agricultural crop production. The rate at which nitrate (NO3 -) is taken up depends partly on the uptake and transpiration of water. Rapid changes in nitrate assimilation, in contrast to other N forms, may serve as a component of the plant stress response to drought because nitrate assimilation may lead to changes in xylem pH. The modulation of xylem sap pH may be relevant for stomata regulation via the delivery of abscisic acid (ABA) to guard cells. In several factorial experiments, we investigated the interactions between nitrate and water availability on nitrate fate in the plant, as well as their possible implications for the early drought-stress response. We monitored the short-term response (2-6 days) of nitrate in biomass, transport to shoot and reduction in Pisum sativum, Hordeum vulgare, Vicia faba, and Nicotiana tabacum and correlated this with sap pH and transpiration rates (TRs). Cultivation on inorganic substrate ensured control over nutrient and water supply and prevented nodulation in legume species. NO3 - content in biomass decreased in most of the species under drought indicating significant decline in NO3 - uptake. Hordeum vulgare had the highest NO3 - concentrations in all organs even under drought and low NO3 - treatment. This species can likely respond much better to the combined adverse effects of low NO3 - and water scarcity. Nitrate reductase activity (NRA) was reduced in both roots and leaves of water deficient (WD) plants in all species except H. vulgare, presumably due to its high NO3 - contents. Further, transient reduction in NO3 - availability had no effect on sap pH. Therefore, it seems unlikely that NRA shifts from shoot root leading to the supposed alkalization of sap. We also did not observe any interactive effects of NO3 - and water deficiency on transpiration. Hence, as long as leaf NO3 - content remains stable, NO3 - availability in soil is not linked to short-term modulation of transpiration.

The pH of the leaf apoplast is critical for the formation of Pseudomonas syringae-induced lesions on leaves of the common bean (Phaseolus vulgaris).

Inoculating a virulent strain of Pseudomonas syringae pv. phaseolicola (Pph) into the leaf of common bean (Phaseolus vulgaris) causes the leaf apoplast to alkalinize. Whether or not this apoplastic pH event facilitates virulence of Pph in interaction with common bean is unclear. For elucidating this topic, (i) Pph colonization of the common bean leaf apoplast, (ii) the formation of bacterial lesions, and (iii) apoplastic sucrose concentration were investigated in relation to the apoplastic leaf pH. For this, the Pph-induced leaf apoplastic alkalinization was attenuated by spray application of either a synthetic auxin or an acidic pH buffer. Apoplastic pH was quantified in planta via microscopy-based pH imaging. Apoplastic washing fluids were extracted to quantify both colonization of bacteria in leaf apoplast and the concentration of apoplastic sucrose. Results reveal that the apoplastic alkalinization facilitated bacterial colonization of the apoplast. Number of colony forming units and area of bacterial lesions were reduced when Pph-induced apoplastic alkalinization was attenuated by foliar application of a synthetic auxin or acidic pH buffer. Application of both agents attenuated the Pph-induced increase of sucrose in the leaf apoplast, which is nutrient for bacteria. Data demonstrate that the Pph-mediated leaf apoplastic alkalinisation favours bacterial colonization.

The role of Arabidopsis ABA receptors from the PYR/PYL/RCAR family in stomatal acclimation and closure signal integration.

Stomata are microscopic pores found on the surfaces of leaves that act to control CO2 uptake and water loss. By integrating information derived from endogenous signals with cues from the surrounding environment, the guard cells, which surround the pore, 'set' the stomatal aperture to suit the prevailing conditions. Much research has concentrated on understanding the rapid intracellular changes that result in immediate changes to the stomatal aperture. In this study, we look instead at how stomata acclimate to longer timescale variations in their environment. We show that the closure-inducing signals abscisic acid (ABA), increased CO2, decreased relative air humidity and darkness each access a unique gene network made up of clusters (or modules) of common cellular processes. However, within these networks some gene clusters are shared amongst all four stimuli. All stimuli modulate the expression of members of the PYR/PYL/RCAR family of ABA receptors. However, they are modulated differentially in a stimulus-specific manner. Of the six members of the PYR/PYL/RCAR family expressed in guard cells, PYL2 is sufficient for guard cell ABA-induced responses, whereas in the responses to CO2, PYL4 and PYL5 are essential. Overall, our work shows the importance of ABA as a central regulator and integrator of long-term changes in stomatal behaviour, including sensitivity, elicited by external signals. Understanding this architecture may aid in breeding crops with improved water and nutrient efficiency.