F-box protein DOR functions as a novel inhibitory factor for abscisic acid-induced stomatal closure under drought stress in Arabidopsis,

Guard cells, which form stoma in leaf epidermis, sense and integrate environmental signals to modulate stomatal aperture in response to diverse conditions. Under drought stress, plants synthesize abscisic acid (ABA), which in turn induces a rapid closing of stoma, to prevent water loss by transpiration. However, many aspects of the molecular mechanism for ABA-mediated stomatal closure are still not understood. Here, we report a novel negative regulator of guard cell ABA signaling, DOR, in Arabidopsis (Arabidopsis thaliana). The DOR gene encodes a putative F-box protein, a member of the S-locus F-box-like family related to AhSLF-S(2) and specifically interacting with ASK14 and CUL1. A null mutation in DOR resulted in a hypersensitive ABA response of stomatal closing and a substantial increase of drought tolerance; in contrast, the transgenic plants overexpressing DOR were more susceptible to the drought stress. DOR is strongly expressed in guard cells and suppressed by ABA treatment, suggesting a negative feedback loop of DOR in ABA responses. Double-mutant analyses of dor with ABA-insensitive mutant abi1-1 showed that abi1-1 is epistatic to dor, but no apparent change of phospholipase Dalpha1 was detected between the wild type and dor. Affymetrix GeneChip analysis showed that DOR likely regulates ABA biosynthesis under drought stress. Taken together, our results demonstrate that DOR acts independent of phospholipase Dalpha1 in an ABA signaling pathway to inhibit the ABA-induced stomatal closure under drought stress.     

Most stomatal closure in woody species under moderate drought can be explained by stomatal responses to leaf turgor

Reduced stomatal conductance (g_s) during soil drought in angiosperms may result from effects of leaf turgor on stomata and/or factors that do not directly depend on leaf turgor, including root‐derived abscisic acid (ABA) signals. To quantify the roles of leaf turgor‐mediated and leaf turgor‐independent mechanisms in g_s decline during drought, we measured drought responses of g_s and water relations in three woody species (almond, grapevine and olive) under a range of conditions designed to generate independent variation in leaf and root turgor, including diurnal variation in evaporative demand and changes in plant hydraulic conductance and leaf osmotic pressure. We then applied these data to a process‐based g_s model and used a novel method to partition observed declines in g_s during drought into contributions from each parameter in the model. Soil drought reduced g_s by 63–84% across species, and the model reproduced these changes well (r² = 0.91, P < 0.0001, n = 44) despite having only a single fitted parameter. Our analysis concluded that responses mediated by leaf turgor could explain over 87% of the observed decline in g_s across species, adding to a growing body of evidence that challenges the root ABA‐centric model of stomatal responses to drought.     

PeCHYR1, a ubiquitin E3 ligase from Populus euphratica,enhances drought tolerance via ABA‐induced stomatal closure by ROS production in Populus

Drought, a primary abiotic stress, seriously affects plant growth and productivity. Stomata play a vital role in regulating gas exchange and drought adaptation. However, limited knowledge exists of the molecular mechanisms underlying stomatal movement in trees. Here, PeCHYR1, a ubiquitin E3 ligase, was isolated from Populus euphratica, a model of stress adaptation in forest trees. PeCHYR1 was preferentially expressed in young leaves and was significantly induced by ABA (abscisic acid) and dehydration treatments. To study the potential biological functions of PeCHYR1, transgenic poplar 84K (Populus alba × Populus glandulosa) plants overexpressing PeCHYR1 were generated. PeCHYR1 overexpression significantly enhanced H₂O₂ production and reduced stomatal aperture. Transgenic lines exhibited increased sensitivity to exogenous ABA and greater drought tolerance than that of WT (wild‐type) controls. Moreover, up‐regulation of PeCHYR1 promoted stomatal closure and decreased transpiration, resulting in strongly elevated WUE (water use efficiency). When exposed to drought stress, transgenic poplar maintained higher photosynthetic activity and biomass accumulation. Taken together, these results suggest that PeCHYR1 plays a crucial role in enhancing drought tolerance via ABA‐induced stomatal closure caused by hydrogen peroxide (H₂O₂) production in transgenic poplar plants.     

Estimating tomato water consumption by sap flow measurement in response to water stress under greenhouse conditions

This research was conducted to determine the water consumption of tomato plants, the effects of water stress on stem sap flow (SF) and its response to climatic factors. SFs in 100% irrigation (T₁), 75% (T₂) and 50% (T₃) of irrigation amount of T₁ were monitored using Dynagage sensors. Compared to T₁, the difference in SF was observed under deficit irrigation in the same climatic conditions on sunny days although there was no apparent difference between T₁ and T₂ on cloudy days. Under T₁, the correlation and regression relationships between SF and climatic factors were analyzed at daytime (6:00–22:00), morning (6:00–14:00) and afternoon (14:00–22:00). Considering daytime, the order of sensitive indicators to SF was VPD?>?LI?>?Ta and LI?>?VPD?>?Ta for the Fall-Winter sunny days and Spring-Summer season, respectively. The water uptake over SFs measured for Fall-Winter and Spring-Summer periods were calculated as 168.65 and 229.18?mm, respectively.     

Assessment and application of the biotin switch technique for examining protein S-nitrosylation under conditions of pharmacologically induced oxidative stress

Protein S-nitrosylation has emerged as a principal mechanism by which nitric oxide exerts biological effects. Among methods for studying protein S-nitrosylation, the biotin switch technique (BST) has rapidly gained popularity because of the ease with which it can detect individual S-nitrosylated (SNO) proteins in biological samples. The identification of SNO sites by the BST relies on the ability of ascorbate to generate a thiol from an S-nitrosothiol, but not from alternatively S-oxidized thiols (e.g. disulfides, sulfenic acids). However, the specificity of this reaction has recently been challenged, prompting several claims that the BST may produce false-positive results and raising concerns about the application of the BST under oxidizing conditions. Here we perform a comparative analysis of the BST using differentially S-oxidized and S-nitrosylated forms of protein tyrosine phosphatase 1B, as well as intact and lysed human embryonic kidney 293 cells treated with S-oxidizing and S-nitrosylating agents, and verify that the assay is highly specific for SNO. Strikingly, exposure of samples to indirect sunlight from a laboratory window resulted in artifactual ascorbate-dependent signals that are likely promoted by the semidehydroascorbate radical; protection from sunlight eliminated the artifact. In contrast, exposure of SNO proteins to a strong ultraviolet light source (SNO photolysis) prior to the BST provided independent verification of assay specificity. By combining BST with photolysis, we have shown that anti-cancer drug-induced oxidative stress facilitates the S-nitrosylation of the major apoptotic effector glyceraldehyde-3-phosphate dehydrogenase. Collectively, these experiments demonstrate that SNO-dependent signaling pathways can be modulated by oxidative conditions and suggest a potential role for S-nitrosylation in antineoplastic drug action.     

Foliar temperature as a tool for quantifying whole-plant transpiration in tree seedlings under laboratory and greenhouse conditions

Plant Ecology - A proper mechanistic understanding of how whole-tree transpiration varies as a function of environmental conditions is essential for predicting how vegetation will respond to...     

Determination of standardized transpiration used as an indirect method for considering stomatal aperture under field conditions

An indirect method for determination of stomata apertures is described. It is based on comparison of two simultaneous curves: the one obtained under natural conditions and the second obtained under standard conditions with minimum air humidity which can be considered as a criterion of stomata apertures.     

Determination of standardized transpiration used as an indirect method for considering stomatal aperture under field conditions

An indirect method for determination of stomata apertures is described. It is based on comparison of two simultaneous curves : the one obtained under natural conditions and the second obtained under standard conditions with minimum air humidity which can be considered as a criterion of stomata apertures.     

Accelerated protein aggregation induced by macrophage migration inhibitory factor under heat stress conditions

Kinetics of thermal aggregation of model protein substrates (glycogen phosphorylase b from rabbit skeletal muscle and yeast alcohol dehydrogenase) were investigated under heat stress conditions (41-48 degrees C) in the presence of macrophage migration inhibitory factor (MIF), a heat-stable hydrophobic protein (12.5 kD). Anti-chaperone MIF activity found by turbidimetry manifests itself in significantly accelerated protein aggregation and increased limiting value of apparent optical absorption at 360 nm and t --> infinity in the sub-stoichiometric range of MIF concentrations. The aggregation kinetics is shown to have cooperative character. Possible reversibility of aggregation after removal of denaturing conditions was demonstrated using alcohol dehydrogenase aggregation at a temperature close to the physiological level (41.5 degrees C). This reversibility is caused by solubility of aggregates and stabilization of oligomeric structure of the substrate as a result of MIF binding to the partially denatured protein. The data suggest that in spite of distinct anti-chaperone effect, the chaperone-like activity of MIF can be observed in the case of heat stress removal and restoration of the system to normal conditions.     

Non-uniform stomatal closure of hybrid poplar clones under light stress determined by scanning electron microscopy and modification of intercellular CO2 concentration

The stomatal distribution, non-uniform stomatal closure, stomatal conductance, and gas-exchange of several hybrid poplar clones under light stress were studied using scanning electron microscopy (SEM) and gas-exchange. Non-uniform stomatal closure was found under natural light stress by SEM, and there was a linear relationship between the stomatal aperture and stomatal conductance. We suggest a formula for modification of intercellular CO₂ concentration, which can restore consideration of stomatal factors leading to midday depression of photosynthesis in some cases. Received: 11 January 1999 / Accepted: 6 April 2000     

Quantitative trait loci for rooting pattern traits of common beans grown under drought stress versus non-stress conditions

Drought is the major abiotic constraint contributing to yield reduction in common bean (Phaseolus vulgaris L.) worldwide. An increasing scarcity of water in the future will make improving adaptation to drought stress a major objective of most crop breeding efforts. Drought avoidance by increased extraction of soil moisture from greater depth under drought conditions is an adaptive mechanism of common bean. A recombinant inbred line population of DOR364?×?BAT477 was evaluated for rooting pattern traits in soil cylinder tubes under soil drying (progressive water stress) and non-stress (well-watered with 80% of field capacity) treatments in a greenhouse. One of the parents, BAT 477, is a deep-rooting genotype while the other parent, DOR 364, is a commercial cultivar in Central America. The recombinant inbred line population expressed quantitative variation and transgressive segregation for ten rooting pattern traits as well as five shoot traits of 48-day-old plants. A mixed model quantitative trait locus (QTL) mapping analysis was carried out using a genetic map constructed with 165 genetic markers that covered 11 linkage groups of the common bean genome. Genotype estimates were calculated from best design and spatial effects model for each trait. A total of 15 putative QTL were identified for seven rooting pattern traits and four shoot traits. The QTL detected were scattered over five of the 11 linkage groups. The QTL detected for all the root traits except total root length and fine root length were main effect QTL and did not interact with the level of water supply. The total root length and fine root length QTL with significant QTL?×?environment interaction only differed in magnitude of effect, and interaction was of a non-crossover type. Other QTL for total root length, fine roots, thick roots, root volume and root biomass were co-localized and also explained relatively more genetic variance. This suggests that the QTL affecting root traits in common beans are based on constitutive expression of genes and that drought avoidance based on deep rooting, longer root length, thicker roots, increasing root length distribution with depth, root volume and root biomass can be used in molecular breeding. The positive alleles for most of the QTL detected in this study were derived from the paternal parent BAT477. The results from the present analyses highlighted the feasibility of marker-aided selection as an alternative to conventional labor-intensive, phenotypic screening of drought avoidance root traits.     

Jasmonic acid transient accumulation is needed for abscisic acid increase in citrus roots under drought stress conditions

Phytohormones are central players in sensing and signaling numerous environmental conditions like drought stress. In this work, an experimental system based on severe drought was established and hormone profiling together with gene expression of key enzymes involved in abscisic acid (ABA) and jasmonic acid (JA) biosynthesis was studied in roots of citrumelo CPB 4475 (a commercial citrus rootstock) plants. JA concentration transiently increased after a few hours of stress, returning to control levels 30 h after the onset of the condition. A more progressive ABA accumulation was observed, with the onset of this increase at the same time or right after the JA transient accumulation. Molecular data suggested that, at least, part of the hormonal regulation takes place at the biosynthetic level. These observations also pointed to a possible involvement of JA on ABA biosynthesis under stress. To test this hypothesis, JA and ABA biosynthesis were chemically inhibited and subsequently phenotypes rescued by the addition of exogenous hormones. Results showed that the early JA accumulation was necessary for the subsequent ABA increase in roots under stress whereas the opposite could not be stated. The model includes a burst of JA in roots of citrus under severe drought stress conditions that leads to a more progressive ABA accumulation that will induce later plant responses. The present work adds a new level of interaction between JA and ABA at the biosynthetic level that together with the previously described interaction between signal transduction cascades of the two hormones would allow plants to fine‐tune specific responses to different stimuli.