Signaling In Higher Plants Research Articles

DPY1 as an osmosensor for drought signaling

Drought stress has been extensively studied for its effect on the downstream signaling cascade and stress-responsive gene expression, but understanding the process has remained elusive. Recently, Zhao et al. identified DROOPY LEAF1 (DPY1) as an osmosensor and revealed a novel mechanism of DPY1-STRESS ACTIVATED PROTEIN KINASE6 (SAPK6)-mediated drought stress signaling in higher plants.

Ion channels in electrical signaling in higher plants

Electrical signals (ESs) of plants occurring under the action of various external factors play an important role in adaptation to changing environmental conditions. The generation of ESs in higher plant cells is associated with the activation of Ca2+, K+, and anion fluxes, as well as changes in the activity of plasma membrane H+-ATPase. In the present review, the molecular nature of ion channels contributing to ESs transmission in higher plants is analyzed based on the comparison of data from molecular-genetic and electrophysiological studies. Based on such characteristics of ion channels as selectivity, activation mechanism, and intracellular and tissue localization, those ion channels that meet the requirements for potential participants of ESs generation were selected from a wide variety of ion channels in higher plants. Analysis of the data of experimental studies performed on mutants with suppressed or enhanced expression of a certain channel gene revealed those channels whose activation contributes to ESs formation. The channels responsible for Ca2+ flux during ESs generation include channels of GLR family, for K+ flux - GORK, for anions - MSL. Consideration of the prospects for further studies suggests the need to combine electrophysiological and genetic approaches along with the analysis of ion concentrations in intact plants within a single study.

Ion Channels in Electrical Signaling in Higher Plants.

Electrical signals (ESs) appearing in plants under the action of various external factors play an important role in adaptation to changing environmental conditions. Generation of ES in higher plant cells is associated with activation of Ca2+, K+, and anion fluxes, as well as with changes in the activity of plasma membrane H+-ATPase. In the present review, molecular nature of the ion channels contributing to ESs transmission in higher plants is analyzed based on comparison of the data from molecular-genetic and electrophysiological studies. Based on such characteristics of ion channels as selectivity, activation mechanism, and intracellular and tissue localization, those ion channels that meet the requirements for potential participation in ES generation were selected from a wide variety of ion channels in higher plants. Analysis of the data of experimental studies performed on mutants with suppressed or enhanced expression of a certain channel gene revealed those channels whose activation contributes to ESs formation. The channels responsible for Ca2+ flux during generation of ESs include channels of the GLR family, for K+ flux - GORK, for anions - MSL. Consideration of the prospects of further studies suggests the need to combine electrophysiological and genetic approaches along with analysis of ion concentrations in intact plants within a single study.

Overexpression of 9-cis-Epoxycarotenoid Dioxygenase Gene, IbNCED1, Negatively Regulates Plant Height in Transgenic Sweet Potato.

Plant height is one of the key agronomic traits for improving the yield of sweet potato. Phytohormones, especially gibberellins (GAs), are crucial to regulate plant height. The enzyme 9-cis-epoxycarotenoid dioxygenase (NCED) is the key enzyme for abscisic acid (ABA) biosynthesis signalling in higher plants. However, its role in regulating plant height has not been reported to date. Here, we cloned a new NCED gene, IbNCED1, from the sweet potato cultivar Jishu26. This gene encoded the 587-amino acid polypeptide containing an NCED superfamily domain. The expression level of IbNCED1 was highest in the stem and the old tissues in the in vitro-grown and field-grown Jishu26, respectively. The expression of IbNCED1 was induced by ABA and GA3. Overexpression of IbNCED1 promoted the accumulation of ABA and inhibited the content of active GA3 and plant height and affected the expression levels of genes involved in the GA metabolic pathway. Exogenous application of GA3 could rescue the dwarf phenotype. In conclusion, we suggest that IbNCED1 regulates plant height and development by controlling the ABA and GA signalling pathways in transgenic sweet potato.

Research status of transcription factors involved in controlling gene expression by nitrate signaling in higher plants

Research status of transcription factors involved in controlling gene expression by nitrate signaling in higher plants

Transport of organic anions in root cells and its role in cell signaling in higher plants

The organic anion balance is critical for metabolic, bioenergetic, and electrochemical processes in plant cells, controlling the quality and quantity of yield and plant stress resistance. Nevertheless, the redistribution and membrane transport of these substances in plant tissues have not been investigated in detail. The mechanism of passive anion efflux from a plant cell through the ion channels has not been established so far. Here, using the patch-clamp technique, we have characterized the ion channel-mediated conductances of ascorbate, malate, gluconate, citrate, fumarate, and pronionate in the root cells of Arabidopsis thaliana, Triticum aestivum, and Helianthus annuus. These conductances showed high permeability to ascorbate, malate, and citrate, as well as low permeability to fumarate, propionate, and gluconate. Anion channel conductances of root cells showed rapid activation kinetics and low potential dependence. They were also inhibited by 9-anthracenecarboxylic acid, suggesting that they belong to the ALMT family of anion channels found only in higher plants. Aequorin chemilu minometry was used to test the effect of organic anions on the Ca2+ signaling in root cells. Among four organic anions tested, only ascorbate induced a significant increase in the cytosolic Ca2+ activity at physiological levels (1 and 10 mM). This effect may underlie the previously unknown functions of exogenous ascorbate related to short- and long-distance signaling in higher plants.

Initiation and amplification of SnRK2 activation in abscisic acid signaling

The phytohormone abscisic acid (ABA) is crucial for plant responses to environmental challenges. The SNF1-regulated protein kinase 2s (SnRK2s) are key components in ABA-receptor coupled core signaling, and are rapidly phosphorylated and activated by ABA. Recent studies have suggested that Raf-like protein kinases (RAFs) participate in ABA-triggered SnRK2 activation. In vitro kinase assays also suggest the existence of autophosphorylation of SnRK2s. Thus, how SnRK2 kinases are quickly activated during ABA signaling still needs to be clarified. Here, we show that both B2 and B3 RAFs directly phosphorylate SnRK2.6 in the kinase activation loop. This transphosphorylation by RAFs is essential for SnRK2 activation. The activated SnRK2s then intermolecularly trans-phosphorylate other SnRK2s that are not yet activated to amplify the response. High-order Arabidopsis mutants lacking multiple B2 and B3 RAFs show ABA hyposensitivity. Our findings reveal a unique initiation and amplification mechanism of SnRK2 activation in ABA signaling in higher plants.

Influence of Local Burning on Difference Reflectance Indices Based on 400-700 nm Wavelengths in Leaves of Pea Seedlings.

Local damage (e.g., burning) induces a variation potential (VP), which is an important electrical signal in higher plants. A VP propagates into undamaged parts of the plant and influences numerous physiological processes, including photosynthesis. Rapidly increasing plant tolerance to stressors is likely to be a result of the physiological changes. Thus, developing methods of revealing VP-induced physiological changes can be used for the remote sensing of plant systemic responses to local damage. Previously, we showed that burning-induced VP influenced a photochemical reflectance index in pea leaves, but the influence of the electrical signals on other reflectance indices was not investigated. In this study, we performed a complex analysis of the influence of VP induction by local burning on difference reflectance indices based on 400–700 nm wavelengths in leaves of pea seedlings. Heat maps of the significance of local burning-induced changes in the reflectance indices and their correlations with photosynthetic parameters were constructed. Large spectral regions with significant changes in these indices after VP induction were revealed. Most changes were strongly correlated to photosynthetic parameters. Some indices, which can be potentially effective for revealing local burning-induced photosynthetic changes, are separately shown. Our results show that difference reflectance indices based on 400–700 nm wavelengths can potentially be used for the remote sensing of plant systemic responses induced by local damages and subsequent propagation of VPs.

A Theoretical Analysis of Relations between Pressure Changes along Xylem Vessels and Propagation of Variation Potential in Higher Plants.

Variation potential (VP) is an important long-distance electrical signal in higher plants that is induced by local damages, influences numerous physiological processes, and participates in plant adaptation to stressors. The transmission of increased hydraulic pressure through xylem vessels is the probable mechanism of VP propagation in plants; however, the rates of the pressure transmission and VP propagation can strongly vary. We analyzed this problem on the basis of a simple mathematical model of the pressure distribution along a xylem vessel, which was approximated by a tube with a pressure gradient. It is assumed that the VP is initiated if the integral over pressure is more than a threshold one, taking into account that the pressure is transiently increased in the initial point of the tube and is kept constant in the terminal point. It was shown that this simple model can well describe the parameters of VP propagation in higher plants, including the increase in time before VP initiation and the decrease in the rate of VP propagation with an increase in the distance from the zone of damage. Considering three types of the pressure dynamics, our model predicts that the velocity of VP propagation can be stimulated by an increase in the length of a plant shoot and also depends on pressure dynamics in the damaged zone. Our results theoretically support the hypothesis about the impact of pressure variations in xylem vessels on VP propagation.

Action and variation potential electrical signals in higher plants

Action and variation potential electrical signals in higher plants

Inactivation of H+-ATPase Participates in the Influence of Variation Potential on Photosynthesis and Respiration in Peas.

Local damage (e.g., burning, heating, or crushing) causes the generation and propagation of a variation potential (VP), which is a unique electrical signal in higher plants. A VP influences numerous physiological processes, with photosynthesis and respiration being important targets. VP generation is based on transient inactivation of H+-ATPase in plasma membrane. In this work, we investigated the participation of this inactivation in the development of VP-induced photosynthetic and respiratory responses. Two- to three-week-old pea seedlings (Pisum sativum L.) and their protoplasts were investigated. Photosynthesis and respiration in intact seedlings were measured using a GFS-3000 gas analyzer, Dual-PAM-100 Pulse-Amplitude-Modulation (PAM)-fluorometer, and a Dual-PAM gas-exchange Cuvette 3010-Dual. Electrical activity was measured using extracellular electrodes. The parameters of photosynthetic light reactions in protoplasts were measured using the Dual-PAM-100; photosynthesis- and respiration-related changes in O2 exchange rate were measured using an Oxygraph Plus System. We found that preliminary changes in the activity of H+-ATPase in the plasma membrane (its inactivation by sodium orthovanadate or activation by fusicoccin) influenced the amplitudes and magnitudes of VP-induced photosynthetic and respiratory responses in intact seedlings. Decreases in H+-ATPase activity (sodium orthovanadate treatment) induced fast decreases in photosynthetic activity and increases in respiration in protoplasts. Thus, our results support the effect of H+-ATPase inactivation on VP-induced photosynthetic and respiratory responses.

Plant Osmotic Stress Signaling: MAPKKKs Meet SnRK2s.

Osmotic stress signaling in higher plants is crucial to cope with abiotic stress. RAF-like MAPKKKs are activated by hyperosmotic stress and activate downstream ABA-unresponsive and ABA-activated SnRK2s, integrating early osmotic stress and ABA signaling cascades. The connection of B2/B3/B4 RAF-like MAPKKKs with SnRK2s is a new paradigm in signal transduction.

A RAF-SnRK2 kinase cascade mediates early osmotic stress signaling in higher plants

Osmoregulation is important for plant growth, development and response to environmental changes. SNF1-related protein kinase 2s (SnRK2s) are quickly activated by osmotic stress and are central components in osmotic stress and abscisic acid (ABA) signaling pathways; however, the upstream components required for SnRK2 activation and early osmotic stress signaling are still unknown. Here, we report a critical role for B2, B3 and B4 subfamilies of Raf-like kinases (RAFs) in early osmotic stress as well as ABA signaling in Arabidopsis thaliana. B2, B3 and B4 RAFs are quickly activated by osmotic stress and are required for phosphorylation and activation of SnRK2s. Analyses of high-order mutants of RAFs reveal critical roles of the RAFs in osmotic stress tolerance and ABA responses as well as in growth and development. Our findings uncover a kinase cascade mediating osmoregulation in higher plants.

Possible amino acid signaling in higher plants

Possible amino acid signaling in higher plants

Molecular machinery of auxin synthesis, secretion, and perception in the unicellular chlorophyte alga Chlorella sorokiniana UTEX 1230.

Indole-3-acetic acid is a ubiquitous small molecule found in all domains of life. It is the predominant and most active auxin in seed plants, where it coordinates a variety of complex growth and development processes. The potential origin of auxin signaling in algae remains a matter of some controversy. In order to clarify the evolutionary context of algal auxin signaling, we undertook a genomic survey to assess whether auxin acts as a signaling molecule in the emerging model chlorophyte Chlorella sorokiniana UTEX 1230. C. sorokiniana produces the auxin indole-3-acetic acid (IAA), which was present in both the cell pellet and in the supernatant at a concentration of ~ 1 nM, and its genome encodes orthologs of genes related to auxin synthesis, transport, and signaling in higher plants. Candidate orthologs for the canonical AUX/IAA signaling pathway were not found; however, auxin-binding protein 1 (ABP1), an alternate auxin receptor, is present and highly conserved at essential auxin binding and zinc coordinating residues. Additionally, candidate orthologs for PIN proteins, responsible for intercellular, vectorial auxin transport in higher plants, were not found, but PILs (PIN-Like) proteins, a recently discovered family that mediates intracellular auxin transport, were identified. The distribution of auxin related gene in this unicellular chlorophyte demonstrates that a core suite of auxin signaling components was present early in the evolution of plants. Understanding the simplified auxin signaling pathways in chlorophytes will aid in understanding phytohormone signaling and crosstalk in seed plants, and in understanding the diversification and integration of developmental signals during the evolution of multicellular plants.

Electrical Signaling, Photosynthesis and Systemic Acquired Acclimation.

Electrical signaling in higher plants is required for the appropriate intracellular and intercellular communication, stress responses, growth and development. In this review, we have focus on recent findings regarding the electrical signaling, as a major regulator of the systemic acquired acclimation (SAA) and the systemic acquired resistance (SAR). The electric signaling on its own cannot confer the required specificity of information to trigger SAA and SAR, therefore, we have also discussed a number of other mechanisms and signaling systems that can operate in combination with electric signaling. We have emphasized the interrelation between ionic mechanism of electrical activity and regulation of photosynthesis, which is intrinsic to a proper induction of SAA and SAR. In a special way, we have summarized the role of non-photochemical quenching and its regulator PsbS. Further, redox status of the cell, calcium and hydraulic waves, hormonal circuits and stomatal aperture regulation have been considered as components of the signaling. Finally, a model of light-dependent mechanisms of electrical signaling propagation has been presented together with the systemic regulation of light-responsive genes encoding both, ion channels and proteins involved in regulation of their activity. Due to space limitations, we have not addressed many other important aspects of hormonal and ROS signaling, which were presented in a number of recent excellent reviews.

Identification of two CiGADs from Caragana intermedia and their transcriptional responses to abiotic stresses and exogenous abscisic acid.

BackgroundGlutamate decarboxylase (GAD), as a key enzyme in the γ -aminobutyric acid (GABA) shunt, catalyzes the decarboxylation of L-glutamate to form GABA. This pathway has attracted much interest because of its roles in carbon and nitrogen metabolism, stress responses, and signaling in higher plants. The aim of this study was to isolate and characterize genes encoding GADs from Caragana intermedia, an important nitrogen-fixing leguminous shrub.MethodsTwo full-length cDNAs encoding GADs (designated as CiGAD1 and CiGAD2) were isolated and characterized. Multiple alignment and phylogenetic analyses were conducted to evaluate their structures and identities to each other and to homologs in other plants. Tissue expression analyses were conducted to evaluate their transcriptional responses to stress (NaCl, ZnSO4, CdCl2, high/low temperature, and dehydration) and exogenous abscisic acid.ResultsThe CiGADs contained the conserved PLP domain and calmodulin (CaM)-binding domain in the C-terminal region. The phylogenetic analysis showed that they were more closely related to the GADs of soybean, another legume, than to GADs of other model plants. According to Southern blotting analysis, CiGAD1 had one copy and CiGAD2-related genes were present as two copies in C. intermedia. In the tissue expression analyses, there were much higher transcript levels of CiGAD2 than CiGAD1 in bark, suggesting that CiGAD2 might play a role in secondary growth of woody plants. Several stress treatments (NaCl, ZnSO4, CdCl2, high/low temperature, and dehydration) significantly increased the transcript levels of both CiGADs, except for CiGAD2 under Cd stress. The CiGAD1 transcript levels strongly increased in response to Zn stress (74.3-fold increase in roots) and heat stress (218.1-fold increase in leaves). The transcript levels of both CiGADs significantly increased as GABA accumulated during a 24-h salt treatment. Abscisic acid was involved in regulating the expression of these two CiGADs under salt stress.DiscussionThis study showed that two CiGADs cloned from C. intermedia are closely related to homologs in another legume, soybean. CiGAD2 expression was much higher than that of CiGAD1 in bark, indicating that CiGAD2 might participate in the process of secondary growth in woody plants. Multiple stresses, interestingly, showed that Zn and heat stresses had the strongest effects on CiGAD1 expression, suggesting that CiGAD1 plays important roles in the responses to Zn and heat stresses. Additionally, these two genes might be involved in ABA dependent pathway during stress. This result provides important information about the role of GADs in woody plants’ responses to environmental stresses.

Evolutionary Analysis of DELLA-Associated Transcriptional Networks.

DELLA proteins are transcriptional regulators present in all land plants which have been shown to modulate the activity of over 100 transcription factors in Arabidopsis, involved in multiple physiological and developmental processes. It has been proposed that DELLAs transduce environmental information to pre-wired transcriptional circuits because their stability is regulated by gibberellins (GAs), whose homeostasis largely depends on environmental signals. The ability of GAs to promote DELLA degradation coincides with the origin of vascular plants, but the presence of DELLAs in other land plants poses at least two questions: what regulatory properties have DELLAs provided to the behavior of transcriptional networks in land plants, and how has the recruitment of DELLAs by GA signaling affected this regulation. To address these issues, we have constructed gene co-expression networks of four different organisms within the green lineage with different properties regarding DELLAs: Arabidopsis thaliana and Solanum lycopersicum (both with GA-regulated DELLA proteins), Physcomitrella patens (with GA-independent DELLA proteins) and Chlamydomonas reinhardtii (a green alga without DELLA), and we have examined the relative evolution of the subnetworks containing the potential DELLA-dependent transcriptomes. Network analysis indicates a relative increase in parameters associated with the degree of interconnectivity in the DELLA-associated subnetworks of land plants, with a stronger effect in species with GA-regulated DELLA proteins. These results suggest that DELLAs may have played a role in the coordination of multiple transcriptional programs along evolution, and the function of DELLAs as regulatory ‘hubs’ became further consolidated after their recruitment by GA signaling in higher plants.

Expression of the chloroplast genome: Modern concepts and experimental approaches

A unique feature of plants is the presence of two extra-nuclear genomes of chloroplasts and mitochondria. The chloroplast (cp) genome is relatively small and contains only 100–120 genes, which encode less than 5% of all proteins required for plastid to function. Expression of the cpDNA retains certain prokaryotic features, such as gene cotranscription within the operon, bacteria-like RNA polymerases and promoters, and 70S ribosomes. However, eukaryotic features also appear in this process, such as the uncoupling of transcription and translation, the involvement of phage-type RNA polymerases, RNA editing, and splicing of the primary transcripts. The interaction between the nucleus (nuclear genome) and cytoplasm (plastid and mitochondrial genomes) during plant development is necessary for proper development and adaptation to the environment. The aim of this review is to disclose the peculiarities of plastid genome expression. The way how the genetic information in chloroplasts is used (transcription, editing, splicing, polyadenylation, and translation) is consequently described. Furthermore, the importance of all expression machinery components in plant life is discussed. Modern approaches for RNA pool studies are described, and the critical points of the nuclear-cytoplasmic interactions in the chloroplast function are revealed. Information about the most important factors of nuclear-cytoplasmic signaling in higher plants (sigma factors and PPR proteins encoded by the nucleus) are reviewed. Thus, the multilevelness and viability of regulating the plastid genome expression in plant cells and the interdependence of the processes in different compartments is proved. A summary of the latest studies of the expression of plastid genome using genetic chips (microarrays and macroarrays) is described. The original results are presented.

Electrical signals in higher plants: Mechanisms of generation and propagation

Local stimulation induces generation and propagation of electric signals in higher plants. Noninvasive stimulus induces an action potential and damaging influences lead to the variation potential. The mechanism of the generation of an action potential is rather complex in nature and is associated with both activation of ion channels (Ca2+, Cl–, and K+) and transient change in the activity of the plasma membrane H+-ATPase. Generation of the variation potential, the duration of which is considerably longer than that of the action potential, is based on transient inactivation of the electrogenic pump; however, passive ion fluxes also contribute to such process, which causes qualitative similarity of the mechanisms of action potential and variation potential generation. Propagation of electrical signals mainly occurs in conducting bundles; thus, transfer of an action potential is associated with vascular parenchyma and sieve elements, while the variation potential is connected to the xylem vessels. The mechanism of the distribution the action potential is similar to nerve impulse transmission, while generation of the variation potential is induced by transfer of a chemical substance, whose propagation is accelerated by a hydraulic wave.