Stress Acclimation Research Articles

Antioxidant and Signaling Role of Plastid-Derived Isoprenoid Quinones and Chromanols.

Plant prenyllipids, especially isoprenoid chromanols and quinols, are very efficient low-molecular-weight lipophilic antioxidants, protecting membranes and storage lipids from reactive oxygen species (ROS). ROS are byproducts of aerobic metabolism that can damage cell components, they are also known to play a role in signaling. Plants are particularly prone to oxidative damage because oxygenic photosynthesis results in O2 formation in their green tissues. In addition, the photosynthetic electron transfer chain is an important source of ROS. Therefore, chloroplasts are the main site of ROS generation in plant cells during the light reactions of photosynthesis, and plastidic antioxidants are crucial to prevent oxidative stress, which occurs when plants are exposed to various types of stress factors, both biotic and abiotic. The increase in antioxidant content during stress acclimation is a common phenomenon. In the present review, we describe the mechanisms of ROS (singlet oxygen, superoxide, hydrogen peroxide and hydroxyl radical) production in chloroplasts in general and during exposure to abiotic stress factors, such as high light, low temperature, drought and salinity. We highlight the dual role of their presence: negative (i.e., lipid peroxidation, pigment and protein oxidation) and positive (i.e., contribution in redox-based physiological processes). Then we provide a summary of current knowledge concerning plastidic prenyllipid antioxidants belonging to isoprenoid chromanols and quinols, as well as their structure, occurrence, biosynthesis and function both in ROS detoxification and signaling.

The rice germin-like protein OsGLP1 participates in acclimation to UV-B radiation.

Exposure to ultraviolet B radiation (UV-B) stress can have serious effects on the growth and development of plants. Germin-like proteins (GLPs) may be involved in different abiotic and biotic stress responses in different plants, but little is known about the role of GLPs in UV-B stress response and acclimation in plants. In the present study, knockout of GLP 8-14 (OsGLP1) using the CRISPR/Cas9 system resulted in mutant rice (Oryza sativa L.) plants (herein called glp1) that exhibited UV-B-dependent formation of lesion mimic in leaves. Moreover, glp1 grown under solar radiation (including UV-B) showed decreased plant height and increased leaf angle, but we observed no significant differences in phenotypes between wild-type (WT) plants and glp1 grown under artificial light lacking UV-B. Fv/Fm, Y (II) and the expression of many genes, based on RNA-seq analysis, related to photosynthesis were also only reduced in glp1, but not in WT, after transfer from a growth cabinet illuminated with artificial white light lacking UV-B to growth under natural sunlight. The genes-associated with flavonoid metabolism as well as UV resistance locus 8 (OsUVR8), phytochrome interacting factor-like 15-like (OsPIF3), pyridoxal 5'-phosphate synthase subunit PDX1.2 (OsPDX1.2), deoxyribodipyrimidine photolyase (OsPHR), and deoxyribodipyrimidine photolyase family protein-like (OsPHRL) exhibited lower expression levels, while higher expression levels of mitogen-activated protein kinase 5-like (OsMPK3), mitogen-activated protein kinase 13-like (OsMPK13), and transcription factor MYB4-like (OsMYB4) were observed in glp1 than in WT after transfer from a growth cabinet illuminated with artificial white light to growth under natural sunlight. Therefore, mutations in OsGLP1 resulted in rice plants more sensitive to UV-B and reduced expression of some genes for UV-B protection, suggesting that OsGLP1 is involved in acclimation to UV-B radiation.

Silicon-Mediated Priming Induces Acclimation to Mild Water-Deficit Stress by Altering Physio-Biochemical Attributes in Wheat Plants.

Water-deficit stress negatively affects seed germination, seedling development, and plant growth by disrupting cellular and metabolic functions, reducing the productivity and yield of field crops. In this study, sodium silicate (SS) has been employed as a seed priming agent for acclimation to mild water-deficit stress by invoking priming memory in wheat plants. In pot experiments, the SS-primed (20, 40, and 60 mM) and non-primed control seeds were allowed to grow under normal and mild water-deficit conditions. Subsequently, known methods were followed for physiological and biochemical studies using flag leaves of 98-day mature wheat plants. The antioxidant and hydrolytic enzymes were upregulated, while proteins, reducing sugars, total sugars, and glycine betaine increased significantly in the flag leaves of wheat plants originated from SS-treated seeds compared to the control under mild water-deficit stress. Significant decreases in the malondialdehyde (MDA) and proline contents suggested a controlled production of reactive oxygen species, which resulted in enhanced cell membrane stability. The SS priming induced a significant enhancement in yield, plant biomass, and 100-grain weight of wheat plants under water-deficit stress. The improvement in the yield parameters indicated the induction of Si-mediated stress acclimation in SS-primed seeds that elicited water-deficit tolerance until the maturity of plants, ensuring sustainable productivity of climate-smart plants.

Sodium nitroprusside mediated priming memory invokes water-deficit stress acclimation in wheat plants through physio-biochemical alterations

AimWater-deficit stress is the most devastating environmental factor that adversely affects plant growth causing yield losses and low crop productivity. In this study, we employed sodium nitroprusside (SNP) as a seed priming agent for the acclimation of water-deficit stress in wheat plants by invoking priming memory. MethodsThe SNP-primed (75, 100, and 125 μM) and non-primed controls were allowed to grow in pots under water deficit and normal conditions. The flag leaves of 98-days mature plants were used for biochemical and physiological studies by following the well-established methods. ResultsThe antioxidant and hydrolytic enzymes were upregulated while reducing sugars, total sugars, and glycine betaine increased significantly in flag leaves of wheat plants originated from SNP-treated seeds compared to control under water deficit stress. However, a significant reduction in MDA and proline contents represented a lesser ROS production which resulted in enhanced cell membrane stability. Consequently, there was a significant enhancement in yield, plant biomass and 100 grains weight of wheat plants under water deficit stress. ConclusionThe improvement in yield parameters indicates the induction of priming memory in SNP-primed seeds which elicit water deficit tolerance till the maturity of plants thus ensures sustainable productivity of wheat.

Attracted to membranes: lipid-binding domains in plants.

Membranes are essential for cells and organelles to function. As membranes are impermeable to most polar and charged molecules, they provide electrochemical energy to transport molecules across and create compartmentalized microenvironments for specific enzymatic and cellular processes. Membranes are also responsible for guided transport of cargoes between organelles and during endo- and exocytosis. In addition, membranes play key roles in cell signaling by hosting receptors and signal transducers and as substrates and products of lipid second messengers. Anionic lipids and their specific interaction with target proteins play an essential role in these processes, which are facilitated by specific lipid-binding domains. Protein crystallography, lipid-binding studies, subcellular localization analyses, and computer modeling have greatly advanced our knowledge over the years of how these domains achieve precision binding and what their function is in signaling and membrane trafficking, as well as in plant development and stress acclimation.

Chloroplast-derived photo-oxidative stress causes changes in H2O2 and EGSH in other subcellular compartments.

Metabolic fluctuations in chloroplasts and mitochondria can trigger retrograde signals to modify nuclear gene expression. Mobile signals likely to be involved are reactive oxygen species (ROS), which can operate protein redox switches by oxidation of specific cysteine residues. Redox buffers, such as the highly reduced glutathione pool, serve as reservoirs of reducing power for several ROS-scavenging and ROS-induced damage repair pathways. Formation of glutathione disulfide and a shift of the glutathione redox potential (EGSH) toward less negative values is considered as hallmark of several stress conditions. Here we used the herbicide methyl viologen (MV) to generate ROS locally in chloroplasts of intact Arabidopsis (Arabidopsis thaliana) seedlings and recorded dynamic changes in EGSH and H2O2 levels with the genetically encoded biosensors Grx1-roGFP2 (for EGSH) and roGFP2-Orp1 (for H2O2) targeted to chloroplasts, the cytosol, or mitochondria. Treatment of seedlings with MV caused rapid oxidation in chloroplasts and, subsequently, in the cytosol and mitochondria. MV-induced oxidation was significantly boosted by illumination with actinic light, and largely abolished by inhibitors of photosynthetic electron transport. MV also induced autonomous oxidation in the mitochondrial matrix in an electron transport chain activity-dependent manner that was milder than the oxidation triggered in chloroplasts by the combination of MV and light. In vivo redox biosensing resolves the spatiotemporal dynamics of compartmental responses to local ROS generation and provides a basis for understanding how compartment-specific redox dynamics might operate in retrograde signaling and stress acclimation in plants.

Transient photoinhibition and photo-oxidative stress as an integral part of stress acclimation and plant development in a dioecious tree adapted to Mediterranean ecosystems.

Mastic trees (Pistacia lentiscus L.) are dioecious perennial plants that are highly adapted to Mediterranean climates but display a high sensitivity to winter periods. In order to understand how sex, leaf phenology and ecological context could condition sensitivity to winter and associated mechanisms to acclimate to these conditions, photoinhibition and photo-oxidative stress markers were examined in mastic trees (P. lentiscus) from a natural population growing in the Garraf Natural Park for a consecutive 12-month period (seasonal study), as well as in three populations naturally growing in the Montseny Natural Park, including the highest altitudes described for this species, during winter (altitudinal study). Results from these studies indicate that both the winter period and higher elevation influenced the degree of photoinhibition, but this was not conditioned by sex. In fact, winter photoinhibition occurred transiently even though it was accompanied by chlorophyll loss and malondialdehyde contents. Stress acclimation was achieved through biochemical adjustments in chloroplasts, characterized by anthocyanin shielding, increased de-epoxidation state of the xanthophyll cycle as well as tocopherol accumulation, and phenological adaptations, the latter allowing a complete resetting of the physiological performance of leaves. Moreover, although females showed higher lipid peroxidation than males during the coldest winter months, at the highest elevation and during flowering in spring, this oxidative stress was mild and transient with no negative consequences for the physiology of plants. It is concluded that evergreen mastic trees acclimate to winter conditions and higher elevations by activation of antioxidant defenses together with phenological adjustments, altogether playing a crucial role in plant survival. Sexual dimorphism in mastic trees appears as a relevant factor when considering sensitivity to photo-oxidative stress in winter and altitudinal conditions.

Morphological plasticity in Chlamydomonas reinhardtii and acclimation to micropollutant stress

Phytoplankton are characterized by a great phenotypic plasticity and amazing morphological variability, both playing a primary role in the acclimation to changing environments. However, there is a knowledge gap concerning the role of algal morphological plasticity in stress responses and acclimation to micropollutants. The present study aims at examining palmelloid colony formation of the green alga Chlamydomonas reinhardtii upon micropollutants exposure. Cells were exposed to four micropollutants (MPs, copper, cadmium, PFOS and paraquat) with different modes of action for a duration of 72 h. Effects of MPs on palmelloid formation, growth and physiological traits (chlorophyll fluorescence, membrane integrity and oxidative stress) were monitored by flow cytometry and fluorescence microscopy. Palmelloid formation was observed upon treatment with the four micropollutants. Number of palmelloid colonies and their size were dependent on MP concentration and exposure duration. Cells reverted to their unicellular lifestyle when colonies were harvested and inoculated in fresh medium indicating that palmelloid formation is a plastic response to micropollutants. No physiological effects of these compounds were observed in cells forming palmelloids. Palmelloid colonies accumulated lower Cd concentration than unicellular C. reinhardtii suggesting that colony formation protects the cells from MPs stress. The results show that colony formation in Chlamydomonas reinhardtii is a stress response strategy activated to face sub-lethal micropollutant concentrations.

Prior exposure of Arabidopsis seedlings to mechanical stress heightens jasmonic acid-mediated defense against necrotrophic pathogens

BackgroundProlonged mechanical stress (MS) causes thigmomorphogenesis, a stress acclimation response associated with increased disease resistance. What remains unclear is if; 1) plants pre-exposed to a short period of repetitive MS can prime defence responses upon subsequent challenge with necrotrophic pathogens, 2) MS mediates plant immunity via jasmonic acid (JA) signalling, and 3) a short period of repetitive MS can cause long-term changes in gene expression resembling a stress-induced memory. To address these points, 10-days old juvenile Arabidopsis seedlings were mechanically stressed for 7-days using a soft brush and subsequently challenged with the necrotrophic pathogens, Alternaria brassicicola, and Botrytis cinerea. Here we assessed how MS impacted structural cell wall appositions, disease symptoms and altered gene expression in response to infection.ResultsThe MS-treated plants exhibited enhanced cell wall appositions and jasmonic acid (JA) accumulation that correlated with a reduction in disease progression compared to unstressed plants. The expression of genes involved in JA signalling, callose deposition, peroxidase and phytoalexin biosynthesis and reactive oxygen species detoxification were hyper-induced 4-days post-infection in MS-treated plants. The loss-of-function in JA signalling mediated by the JA-insensitive coronatine-insensitive 1 (coi1) mutant impaired the hyper-induction of defense gene expression and promoted pathogen proliferation in MS-treated plants subject to infection. The basal expression level of PATHOGENESIS-RELATED GENE 1 and PLANT DEFENSIN 1.2 defense marker genes were constitutively upregulated in rosette leaves for 5-days post-MS, as well as in naïve cauline leaves that differentiated from the inflorescence meristem well after ceasing MS.ConclusionThis study reveals that exposure of juvenile Arabidopsis plants to a short repetitive period of MS can alter gene expression and prime plant resistance upon subsequent challenge with necrotrophic pathogens via the JA-mediated COI1 signalling pathway. MS may facilitate a stress-induced memory to modulate the plant’s response to future stress encounters. These data advance our understanding of how MS primes plant immunity against necrotrophic pathogens and how that could be utilised in sustainable agricultural practices.

Rapid Reduction of Herbicide Susceptibility in Junglerice by Recurrent Selection with Sublethal Dose of Herbicides and Heat Stress

Global climate change, specifically rising temperature, can alter the molecular physiology of weedy plants. These changes affect herbicide efficacy and weed management. This research aimed to investigate the combined effect of heat stress (HS) and sublethal doses of herbicides (four active ingredients) on adaptive gene expression and efficacy of herbicide on Echinochloa colona (L.) Link (junglerice). Three factors were evaluated; factor A was E. colona generation (G0-original population from susceptible standard; G1 and G2 were progenies of recurrent selection), factor B was herbicide treatment (florpyrauxifen-benzyl, glufosinate-ammonium, imazethapyr, quinclorac and nontreated check) and factor C was HS (30 and 45 °C). The herbicides were applied at 0.125× the recommended dose. Recurrent exposure to HS, combined with sublethal doses of herbicides, favors the selection of plants less susceptible to the herbicide. Upregulation of defense (antioxidant) genes (APX: Ascorbate peroxidase), herbicide detoxification genes (CYP450 family: Cytochrome P450), stress acclimation genes (HSP: Heat shock protein, TPP: Trehalose phosphate phosphatase and TPS: Trehalose phosphate synthase) and genes related to herbicide conjugation (UGT: UDP Glucosyltransferase) was significant. The positive regulation of these genes may promote increased tolerance of E. colona to these herbicides.

Phytochrome type B family: The abiotic stress responses signaller in plants

AbstractPhotoreceptors are primarily known as key photomorphogenic modulators of various physiological events during plant development. Although there are different groups of photoreceptors, the phytochrome B (phyB) family mediates developmental responses in a wide range of plant species, from seed germination to flowering. In addition, these molecules also regulate abiotic stress acclimation responses, such as salinity, drought, low/high temperature, high light and heavy metals. The signalling pathways mediated by phyB could enhance plant resistance to environmental stresses, as phyB photoreceptors reduce leaf transpiration, increase the antioxidant system, enhance protective pigments and increase the expression of genes related to plant stress acclimation. Thus, the elucidation of positive or negative roles for phyB in these stress tolerance characteristics would provide essential knowledge for genetic engineering, improving plant growth and development in critical environments. In this review, we cover the main findings on how the phyB family works to modulate abiotic stress by discussing biochemical and molecular aspects of the underlying mechanisms operated by these photoreceptors.

Transcriptional memories mediate the plasticity of cold stress responses to enable morphological acclimation in Brachypodium distachyon.

Summary Plants that successfully acclimate to stress can resume growth under stressful conditions. The grass Brachypodium distachyon can grow a cold‐adaptive morphology during cold acclimation. Studies on transcriptional memory (TM) have revealed that plants can be primed for stress by adjusting their transcriptional responses, but the function of TM in stress acclimation is not well understood. We investigated the function of TM during cold acclimation in B. distachyon.Quantitative polymerase chain reaction (qPCR), RNA‐seq and chromatin immunoprecipitation qPCR analyses were performed on plants exposed to repeated episodes of cold to characterize the presence and stability of TM during the stress and growth responses of cold acclimation.Transcriptional memory mainly dampened stress responses as growth resumed and as B. distachyon became habituated to cold stress. Although permanent on vernalization gene VRN1, TMs were short‐term and reversible on cold‐stress genes. Growing under cold conditions also coincided with the acquisition of new and targeted cold‐induced transcriptional responses.Overall, TM provided plasticity to cold stress responses during cold acclimation in B. distachyon, leading to stress habituation, acquired stress responses, and resumed growth. Our study shows that chromatin‐associated TMs are involved in tuning plant responses to environmental change and, as such, regulate both stress and developmental components that characterize cold‐climate adaptation in B. distachyon.

Remodeling of Root Growth Under Combined Arsenic and Hypoxia Stress Is Linked to Nutrient Deprivation.

Root architecture responds to environmental stress. Stress-induced metabolic and nutritional changes affect the endogenous root development program. Transcriptional and translational changes realize the switch between stem cell proliferation and cell differentiation, lateral root or root hair formation and root functionality for stress acclimation. The current work explores the effects of stress combination of arsenic toxicity (As) and hypoxia (Hpx) on root development in Arabidopsis thaliana. As revealed previously, combined As and Hpx treatment leads to severe nutritional disorder evident from deregulation of root transcriptome and plant mineral contents. Both As and Hpx were identified to pose stress-specific constraints on root development that lead to unique root growth phenotype under their combination. Besides inhibition of root apical meristem (RAM) activity under all stresses, As induced lateral root growth while root hair density and lengths were strongly increased by Hpx and HpxAs-treatments.A dual stimulation of phosphate (Pi)-starvation response was observed for HpxAs-treated plant roots; however, the response under HpxAs aligned more with Hpx than As. Transcriptional evidence along with biochemical data suggests involvement of PHOSPHATE STARVATION RESPONSE 1; PHR1-dependent systemic signaling. Pi metabolism-related transcripts in close association with cellular iron homeostasis modulate root development under HpxAs. Early redox potential changes in meristematic cells, differential ROS accumulation in root hair zone cell layers and strong deregulation of NADPH oxidases, NADPH-dependent oxidoreductases and peroxidases signify a role of redox and ROS signaling in root architecture remodeling under HpxAs. Differential aquaporin expression suggests transmembrane ROS transport to regulate root hair induction and growth. Reorganization of energy metabolism through NO-dependent alternate oxidase, lactate fermentation, and phosphofructokinase seems crucial under HpxAs. TOR and SnRK-signaling network components were potentially involved in control of sustainable utilization of available energy reserves for root hair growth under combined stress as well as recovery on reaeration. Findings are discussed in context of combined stress-induced signaling in regulation of root development in contrast to As and Hpx alone.

Plastidial (p)ppGpp Synthesis by the Ca2+-Dependent RelA-SpoT Homolog Regulates the Adaptation of Chloroplast Gene Expression to Darkness in Arabidopsis.

In bacteria, the hyper-phosphorylated nucleotide, guanosine 3',5'-bis(pyrophosphate) (ppGpp), functions as a secondary messenger under stringent conditions. ppGpp levels are controlled by two distinct enzymes, namely RelA and SpoT, in Escherichia coli. RelA-SpoT homologs (RSHs) are also conserved in plants where they function in the plastids. The model plant Arabidopsis thaliana contains four RSHs: RSH1, RSH2, RSH3 and Ca2+-dependent RSH (CRSH). Genetic characterizations of RSH1, RSH2 and RSH3 were undertaken, which showed that the ppGpp-dependent plastidial stringent response significantly influences plant growth and stress acclimation. However, the physiological significance of CRSH-dependent ppGpp synthesis remains unclear, as no crsh-null mutant has been available. Here, to investigate the function of CRSH, a crsh-knockout mutant of Arabidopsis was constructed using a site-specific gene-editing technique, and its phenotype was characterized. A transient increase in ppGpp was observed for 30 min in the wild type (WT) after the light-to-dark transition, but this increase was not observed in the crsh mutant. Similar analyses were performed with the rsh2-rsh3 double and rsh1-rsh2-rsh3 triple mutants and showed that the transient increments of ppGpp in the mutants were higher than those in the WT. The increase in ppGpp in the WT and rsh2 rsh3 accompanied decrements in the mRNA levels of some plastidial genes transcribed by the plastid-encoded plastid RNA polymerase. These results indicate that the transient increase in ppGpp at night is due to CRSH-dependent ppGpp synthesis and that the ppGpp level is maintained by the hydrolytic activities of RSH1, RSH2 and RSH3 to accustom plastidial gene expression to darkness.

Recurrent Selection by Herbicide Sublethal Dose and Drought Stress Results in Rapid Reduction of Herbicide Sensitivity in Junglerice

Echinochloa colona (junglerice) is a problematic global weed for many crops, primarily controlled with herbicides. Drought stress alters the overall plant physiology and reduces herbicide efficacy. This research aimed to study the joint effect of drought stress (DS) and recurrent selection with sublethal dose of herbicide on adaptive gene expression and herbicide efficacy on E. colona. Three factors were evaluated: (A) E. colona generation (G0, original population from susceptible standard; G1 and G2, progenies of recurrent selection); (B) herbicide treatment (florpyrauxifen-benzyl, 0.25×; glyphosate, 0.125×; quinclorac, 0.125× the recommended dose; and nontreated check); (C) DS (50% and 100% field capacity). Recurrent exposure to sublethal herbicide dose, combined with drought stress, favors the selection of plants less susceptible to the herbicide. Upregulation of defense (antioxidant) genes (APX: ascorbate peroxidase), herbicide detoxification genes (CYP450 family: cytochrome P450), stress acclimation genes (HSP: heat-shock protein, TPP: trehalose phosphate phosphatase, and TPS: trehalose phosphate synthase), and genes related to herbicide conjugation (UGT: UDP glucosyltransferase) in the G2 population was significant. Recurrent exposure to sublethal herbicide dose under drought stress reduces junglerice sensitivity to herbicide, seemingly due to “imprinted” upregulation of metabolic and protection genes in response to these stresses.

Large geographic variability in the resistance of corals to thermal stress

AbstractAimPredictions for the future of coral reefs are largely based on thermal exposure and poorly account for potential geographic variation in biological sensitivity to thermal stress. Without accounting for complex sensitivity responses, simple climate exposure models and associated predictions may lead to poor estimates of future coral survival and lead to policies that fail to identify and implement the most appropriate interventions. To begin filling this gap, we evaluated a number of attributes of coral taxa and communities that are predicted to influence coral resistance to thermal stress over a large geographic range.LocationWestern Indo‐Pacific and Central Indo‐Pacific Ocean Realms.Major taxa studiedZooxanthellate Scleractinia – hard corals.MethodsWe evaluated the geographic variability of coral resistance to thermal stress as the ratio of thermal exposure and sensitivity in 12 countries during the 2016 global‐bleaching event. Thermal exposure was estimated by two metrics: (a) historical excess summer heat (cumulative thermal anomaly, CTA), and (b) a multivariate index of sea‐surface temperature (SST), light, and water flow (climate exposure, CE). Sensitivity was estimated for 226 sites using coordinated bleaching observations and underwater surveys of coral communities. We then evaluated coral resistance to thermal stress using 48 generalized linear mixed models (GLMMs) to compare the potential influences of geography, historical SST variation, coral cover and coral richness.ResultsGeographic faunal provinces and ecoregions were the strongest predictors of coral resistance to thermal stress, with sites in the Australian, Indonesian and Fiji‐Caroline Islands coral provinces having higher resistance to thermal stress than Africa‐India and Japan‐Vietnam provinces. Ecoregions also showed strong gradients in resistance with highest resistance to thermal stress in the western Pacific and Coral Triangle and lower resistance in the surrounding ecoregions. A more detailed evaluation of Coral Triangle and non‐Coral Triangle sites found higher resistance to thermal stress within the Coral Triangle, associated with c. 2.5 times more recent historical thermal anomalies and more centralized, warmer, and cool‐water skew SST distributions, than in non‐Coral Triangle sites. Our findings identify the importance of environmental history and geographic context in future predictions of bleaching, and identify some potential drivers of coral resistance to thermal stress.Main conclusionsSimple threshold models of heat stress and coral acclimation are commonly used to predict the future of coral reefs. Here and elsewhere we show that large‐scale responses of coral communities to heat stress are geographically variable and associated with differential environmental stresses and histories.

Auxin Homeostasis and Distribution of the Auxin Efflux Carrier PIN2 Require Vacuolar NHX-Type Cation/H+ Antiporter Activity.

The Arabidopsis vacuolar Na+/H+ transporters (NHXs) are important regulators of intracellular pH, Na+ and K+ homeostasis and necessary for normal plant growth, development, and stress acclimation. Arabidopsis contains four vacuolar NHX isoforms known as AtNHX1 to AtNHX4. The quadruple knockout nhx1nhx2nhx3nhx4, lacking any vacuolar NHX-type antiporter activity, displayed auxin-related phenotypes including loss of apical dominance, reduced root growth, impaired gravitropism and less sensitivity to exogenous IAA and NAA, but not to 2,4-D. In nhx1nhx2nhx3nhx4, the abundance of the auxin efflux carrier PIN2, but not PIN1, was drastically reduced at the plasma membrane and was concomitant with an increase in PIN2 labeled intracellular vesicles. Intracellular trafficking to the vacuole was also delayed in the mutant. Measurements of free IAA content and imaging of the auxin sensor DII-Venus, suggest that auxin accumulates in root tips of nhx1nhx2nhx3nhx4. Collectively, our results indicate that vacuolar NHX dependent cation/H+ antiport activity is needed for proper auxin homeostasis, likely by affecting intracellular trafficking and distribution of the PIN2 efflux carrier.

Transcriptional and Metabolic Profiling of Potato Plants Expressing a Plastid-Targeted Electron Shuttle Reveal Modulation of Genes Associated to Drought Tolerance by Chloroplast Redox Poise.

Water limitation represents the main environmental constraint affecting crop yield worldwide. Photosynthesis is a primary drought target, resulting in over-reduction of the photosynthetic electron transport chain and increased production of reactive oxygen species in plastids. Manipulation of chloroplast electron distribution by introducing alternative electron transport sinks has been shown to increase plant tolerance to multiple environmental challenges including hydric stress, suggesting that a similar strategy could be used to improve drought tolerance in crops. We show herein that the expression of the cyanobacterial electron shuttle flavodoxin in potato chloroplasts protected photosynthetic activities even at a pre-symptomatic stage of drought. Transcriptional and metabolic profiling revealed an attenuated response to the adverse condition in flavodoxin-expressing plants, correlating with their increased stress tolerance. Interestingly, 5–6% of leaf-expressed genes were affected by flavodoxin in the absence of drought, representing pathways modulated by chloroplast redox status during normal growth. About 300 of these genes potentially contribute to stress acclimation as their modulation by flavodoxin proceeds in the same direction as their drought response in wild-type plants. Tuber yield losses under chronic water limitation were mitigated in flavodoxin-expressing plants, indicating that the flavoprotein has the potential to improve major agronomic traits in potato.

Foliar absorption coefficient derived from reflectance spectra: A gauge of the efficiency of in situ light-capture by different pigment groups

The absorption of Photosynthetically Active Radiation (PAR) by different foliar pigments defines the amount of energy available for photosynthesis and also the need for photoprotection. Both characteristics reveal essential information about productivity, development, and stress acclimation of plants. Here we present an approach for the estimation of the efficiency by three foliar pigment groups (chlorophylls, carotenoids, and anthocyanins) at capturing light, via the absorption coefficient derived from leaf reflectance spectra. The absorption coefficient (and hence light capture efficiency) of the pigment is quantitatively related to the ratio of light absorbed by each pigment group over the total amount of light absorbed by the leaf. The proposed approach allows discerning the contribution of pigment groups to the overall light absorption, despite the strong interference by other pigments with overlapping absorption spectra. For photosynthetic pigments, like chlorophylls, this is indicative of the energy captured for photosynthesis and hence of potential plant productivity. For photoprotective pigments, like anthocyanins or secondary carotenoids, it gives information about the spectral ranges where their optical screening works best and their screening capacity. In addition, the approach allows the selection of optimal spectral bands where different pigments operate. Such information improves our understanding of the phenological, physiological and photosynthetic dynamics of plants over space and through time, useful for developing better monitoring and management strategies.

Identification of Novel Inhibitors of a Plant Group A Protein Phosphatase Type 2C Using a Combined In Silico and Biochemical Approach.

Type 2C protein phosphatases (PP2Cs) of group A play a significant role in the regulation of various processes in plants including growth, development, ion transport, and stress acclimation. In this study, we selected potential PP2C group A inhibitors using a structure-based virtual screening method followed by biochemical and in vitro validation. Over twenty million chemical compounds from the ZINC database were used for docking studies. The precision of the calculations was increased by an induced-fit docking protocol and the molecular mechanics/generalized Born surface area (MM/GBSA) method, which yielded approximate values for the binding energy of the protein-ligand complex. After clustering and ranking their activity, the top-ranking compounds were tested against PP2C group A members in vitro and their in vivo activity was also explored. Phosphatase activity assays identified two compounds with significant inhibitory activity against ABI1 protein ranging from around 57 to 91% at a concentration of 100 μM. Importantly, this in vitro activity correlated well with in vivo inhibition of seed germination, as expected for PP2C inhibitors. The results should promote the design of novel inhibitors with improved potency against ABI1-like and other PP2Cs that might be used in agriculture for the protection of crops against stress.