ABSTRACT
Partial resistance to multiple biotrophic fungal pathogens in wheat (Triticum aestivum L.) is conferred by a variant of the Lr67 gene, which encodes a hexose-proton symporter. Two mutations (G144R and V387L) differentiate the resistant and susceptible protein variants (Lr67res and Lr67sus). Lr67res lacks sugar transport capability and was associated with anion transporter-like properties when expressed in Xenopus laevis oocytes. Here, we extended this functional characterization to include yeast and in planta studies. The Lr67res allele, but not Lr67sus, induced sensitivity to ions in yeast (including NaCl, LiCl, and KI), which is consistent with our previous observations that Lr67res expression in oocytes induces novel ion fluxes. We demonstrate that another naturally occurring single amino acid variant in wheat, containing only the Lr67G144R mutation, confers rust resistance. Transgenic barley plants expressing the orthologous HvSTP13 gene carrying the G144R and V387L mutations were also more resistant to Puccinia hordei infection. NaCl treatment of pot-grown adult wheat plants with the Lr67res allele induced leaf tip necrosis and partial leaf rust resistance. An Lr67res-like function can be introduced into orthologous plant hexose transporters via single amino acid mutation, highlighting the strong possibility of generating disease resistance in other crops, especially with gene editing.
Subject(s)
Disease Resistance , Hordeum , Plant Diseases , Plant Proteins , Triticum , Triticum/genetics , Triticum/microbiology , Plant Diseases/microbiology , Plant Diseases/genetics , Plant Diseases/immunology , Plant Proteins/genetics , Plant Proteins/metabolism , Disease Resistance/genetics , Hordeum/genetics , Hordeum/microbiology , Basidiomycota/physiology , Polymorphism, Genetic , Monosaccharide Transport Proteins/genetics , Monosaccharide Transport Proteins/metabolism , Plants, Genetically Modified/geneticsABSTRACT
Many disease resistance genes in wheat (Triticum aestivum L.) confer strong resistance to specific pathogen races or strains, and only a small number of genes confer multipathogen resistance. The Leaf rust resistance 67 (Lr67) gene fits into the latter category as it confers partial resistance to multiple biotrophic fungal pathogens in wheat and encodes a Sugar Transport Protein 13 (STP13) family hexose-proton symporter variant. Two mutations (G144R, V387L) in the resistant variant, Lr67res, differentiate it from the susceptible Lr67sus variant. The molecular function of the Lr67res protein is not understood, and this study aimed to broaden our knowledge on this topic. Biophysical analysis of the wheat Lr67sus and Lr67res protein variants was performed using Xenopus laevis oocytes as a heterologous expression system. Oocytes injected with Lr67sus displayed properties typically associated with proton-coupled sugar transport proteins-glucose-dependent inward currents, a Km of 110 ± 10 µM glucose, and a substrate selectivity permitting the transport of pentoses and hexoses. By contrast, Lr67res induced much larger sugar-independent inward currents in oocytes, implicating an alternative function. Since Lr67res is a mutated hexose-proton symporter, the possibility of protons underlying these currents was investigated but rejected. Instead, currents in Lr67res oocytes appeared to be dominated by anions. This conclusion was supported by electrophysiology and 36Cl- uptake studies and the similarities with oocytes expressing the known chloride channel from Torpedo marmorata, TmClC-0. This study provides insights into the function of an important disease resistance gene in wheat, which can be used to determine how this gene variant underpins disease resistance in planta.
Subject(s)
Disease Resistance , Triticum , Disease Resistance/genetics , Triticum/metabolism , Chlorine/metabolism , Radioisotopes/metabolism , Monosaccharide Transport Proteins/genetics , Protons , Oocytes/metabolism , Hexoses/metabolism , Glucose , Sugars , Plant Diseases/genetics , Plant Diseases/microbiologyABSTRACT
Growing at either 15 or 25°C, roots of Arabidopsis thaliana, Columbia accession, produce cells at the same rate and have growth zones of the same length. To determine whether this constancy is related to energetics, we measured oxygen uptake by means of a vibrating oxygen-selective electrode. Concomitantly, the spatial distribution of elongation was measured kinematically, delineating meristem and elongation zone. All seedlings were germinated, grown, and measured at a given temperature (15 or 25°C). Columbia was compared to lines where cell production rate roughly doubles between 15 and 25°C: Landsberg and two Columbia mutants, er-105 and ahk3-3. For all genotypes and temperatures, oxygen uptake rate at any position was highest at the root cap, where mitochondrial density was maximal, based on the fluorescence of a reporter. Uptake rate declined through the meristem to plateau within the elongation zone. For oxygen uptake rate integrated over a zone, the meristem had steady-state Q10 values ranging from 0.7 to 2.1; by contrast, the elongation zone had values ranging from 2.6 to 3.3, implying that this zone exerts a greater respiratory demand. These results highlight a substantial energy consumption by the root cap, perhaps helpful for maintaining hypoxia in stem cells, and suggest that rapid elongation is metabolically more costly than is cell division.
Subject(s)
Arabidopsis Proteins , Arabidopsis , Arabidopsis/genetics , Arabidopsis Proteins/genetics , Meristem , Oxygen , Plant Roots , TemperatureABSTRACT
In plants, calcineurin B-like (CBL) proteins and their interacting protein kinases (CIPK) form functional complexes that transduce downstream signals to membrane effectors assisting in their adaptation to adverse environmental conditions. This study addresses the issue of the physiological role of CIPK9 in adaptive responses to salinity, osmotic stress, and K+ deficiency in rice plants. Whole-plant physiological studies revealed that Oscipk9 rice mutant lacks a functional CIPK9 gene and displayed a mildly stronger phenotype, both under saline and osmotic stress conditions. The reported difference was attributed to the ability of Oscipk9 to maintain significantly higher stomatal conductance (thus, a greater carbon gain). Oscipk9 plants contained much less K+ in their tissues, implying the role of CIPK9 in K+ acquisition and homeostasis in rice. Oscipk9 roots also showed hypersensitivity to ROS under conditions of low K+ availability suggesting an important role of H2O2 signalling as a component of plant adaptive responses to a low-K environment. The likely mechanistic basis of above physiological responses is discussed.
ABSTRACT
Soybean (Glycine max) yields are threatened by multiple stresses including soil salinity. GmSALT3 (a cation-proton exchanger protein) confers net shoot exclusion for both Na+ and Cl- and improves salt tolerance of soybean; however, how the ER-localized GmSALT3 achieves this is unknown. Here, GmSALT3's function was investigated in heterologous systems and near isogenic lines that contained the full-length GmSALT3 (NIL-T; salt-tolerant) or a truncated transcript Gmsalt3 (NIL-S; salt-sensitive). GmSALT3 restored growth of K+ -uptake-defective Escherichia coli and contributed towards net influx and accumulation of Na+ , K+ and Cl- in Xenopus laevis oocytes, while Gmsalt3 was non-functional. Time-course analysis of NILs confirmed shoot Cl- exclusion occurs distinctly from Na+ exclusion. Grafting showed that shoot Na+ exclusion occurs via a root xylem-based mechanism; in contrast, NIL-T plants exhibited significantly greater Cl- content in both the stem xylem and phloem sap compared to NIL-S, indicating that shoot Cl- exclusion likely depends upon novel phloem-based Cl- recirculation. NIL-T shoots grafted on NIL-S roots contained low shoot Cl- , which confirmed that Cl- recirculation is dependent on the presence of GmSALT3 in shoots. Overall, these findings provide new insights on GmSALT3's impact on salinity tolerance and reveal a novel mechanism for shoot Cl- exclusion in plants.
Subject(s)
Chlorides/metabolism , Glycine max/metabolism , Plant Leaves/metabolism , Plant Proteins/metabolism , Plant Roots/metabolism , Plant Shoots/metabolism , Sodium/metabolism , Animals , Escherichia coli , Ion Transport , Microscopy, Electron, Transmission , Oocytes , Organisms, Genetically Modified , Plant Leaves/physiology , Plant Proteins/physiology , Plant Roots/physiology , Plant Shoots/physiology , Potassium/metabolism , Salt Tolerance , Glycine max/physiology , Xenopus laevis , Xylem/metabolismABSTRACT
Malate exudation through wheat (Triticum aestivum L) aluminium-activated malate transporter 1 (TaALMT1) confers Al3+ tolerance at low pH, but is also activated by alkaline pH, and is regulated by and facilitates significant transport of gamma-aminobutyric acid (GABA, a zwitterionic buffer). Therefore, TaALMT1 may facilitate acidification of an alkaline rhizosphere by promoting exudation of both malate and GABA. Here, the performance of wheat near isogenic lines ET8 (Al+3 -tolerant, high TaALMT1 expression) and ES8 (Al+3 -sensitive, low TaALMT1 expression) are compared. Root growth (at 5 weeks) was higher for ET8 than ES8 at pH 9. ET8 roots exuded more malate and GABA at high pH and acidified the rhizosphere more rapidly. GABA and malate exudation was enhanced at high pH by the addition of aluminate in both ET8 and transgenic barley expressing TaALMT1. Xenopus laevis oocytes expressing TaALMT1 acidified an alkaline media more rapidly than controls corresponding to higher GABA efflux. TaALMT1 expression did not change under alkaline conditions but key genes involved in GABA turnover changed in accordance with a high rate of GABA synthesis. We propose that TaALMT1 plays a role in alkaline tolerance by exuding malate and GABA, possibly coupled to proton efflux.
Subject(s)
GABA Plasma Membrane Transport Proteins/metabolism , Malates/metabolism , Organic Anion Transporters/metabolism , Plant Proteins/metabolism , Triticum/metabolism , gamma-Aminobutyric Acid/metabolism , Animals , Animals, Genetically Modified , Chlorophyll/metabolism , GABA Plasma Membrane Transport Proteins/physiology , Hordeum , Hydrogen-Ion Concentration , Oocytes , Organic Anion Transporters/physiology , Plant Leaves/metabolism , Plant Proteins/physiology , Plant Roots/metabolism , Plant Roots/physiology , Plants, Genetically Modified , Rhizosphere , Seedlings/metabolism , Seedlings/physiology , Stress, Physiological , Triticum/physiology , XenopusABSTRACT
In rice, the OsHKT1;5 gene has been reported to be a critical determinant of salt tolerance. This gene is harbored by the SKC1 locus, and its role was attributed to Na+ unloading from the xylem. No direct evidence, however, was provided in previous studies. Also, the reported function of SKC1 on the loading and delivery of K+ to the shoot remains to be explained. In this work, we used an electrophysiological approach to compare the kinetics of Na+ uptake by root xylem parenchyma cells using wild type (WT) and NIL(SKC1) plants. Our data showed that Na+ reabsorption was observed in WT, but not NIL(SKC1) plants, thus questioning the functional role of HKT1;5 as a transporter operating in the direct Na+ removal from the xylem. Instead, changes in the expression level of HKT1;5 altered the activity of membrane transporters involved in K+ and Ca2+ acquisition and homeostasis in the rice epidermis and stele, explaining the observed phenotype. We conclude that the role of HKT1;5 in plant salinity tolerance cannot be attributed to merely reducing Na+ concentration in the xylem sap but triggers a complex feedback regulation of activities of other transporters involved in the maintenance of plant ionic homeostasis and signaling under stress conditions.
Subject(s)
Cation Transport Proteins/genetics , Homeostasis/genetics , Oryza/genetics , Oryza/metabolism , Plant Proteins/genetics , Potassium/metabolism , Sodium/metabolism , Symporters/genetics , Gene Expression Regulation, Plant/genetics , Genes, Plant/genetics , Homeostasis/physiology , Membrane Transport Proteins/genetics , Mesophyll Cells/metabolism , Mesophyll Cells/physiology , Phenotype , Plant Roots/genetics , Plant Roots/metabolism , Plant Roots/physiology , Plant Shoots/genetics , Plant Shoots/metabolism , Plant Shoots/physiology , Salt Tolerance/genetics , Salt Tolerance/physiology , Salt-Tolerant Plants/genetics , Salt-Tolerant Plants/metabolism , Salt-Tolerant Plants/physiology , Stress, Physiological/genetics , Xylem/genetics , Xylem/metabolism , Xylem/physiologyABSTRACT
Improving salinity tolerance in the most widely cultivated cereal, bread wheat (Triticum aestivum L.), is essential to increase grain yields on saline agricultural lands. A Portuguese landrace, Mocho de Espiga Branca accumulates up to sixfold greater leaf and sheath sodium (Na+ ) than two Australian cultivars, Gladius and Scout, under salt stress in hydroponics. Despite high leaf and sheath Na+ concentrations, Mocho de Espiga Branca maintained similar salinity tolerance compared to Gladius and Scout. A naturally occurring single nucleotide substitution was identified in the gene encoding a major Na+ transporter TaHKT1;5-D in Mocho de Espiga Branca, which resulted in a L190P amino acid residue variation. This variant prevents Mocho de Espiga Branca from retrieving Na+ from the root xylem leading to a high shoot Na+ concentration. The identification of the tissue-tolerant Mocho de Espiga Branca will accelerate the development of more elite salt-tolerant bread wheat cultivars.
Subject(s)
Plant Proteins/genetics , Plant Shoots/metabolism , Sodium/metabolism , Triticum/genetics , Triticum/metabolism , Animals , Female , Gene Expression Regulation, Plant , Models, Molecular , Oocytes/metabolism , Plant Leaves/genetics , Plant Leaves/metabolism , Plant Proteins/chemistry , Plant Proteins/metabolism , Plant Shoots/genetics , Polymorphism, Single Nucleotide , Potassium-Hydrogen Antiporters/chemistry , Potassium-Hydrogen Antiporters/genetics , Potassium-Hydrogen Antiporters/metabolism , Salt Tolerance/genetics , Xenopus laevis , Xylem/genetics , Xylem/metabolismABSTRACT
Agriculture is expanding into regions that are affected by salinity. This review considers the energetic costs of salinity tolerance in crop plants and provides a framework for a quantitative assessment of costs. Different sources of energy, and modifications of root system architecture that would maximize water vs ion uptake are addressed. Energy requirements for transport of salt (NaCl) to leaf vacuoles for osmotic adjustment could be small if there are no substantial leaks back across plasma membrane and tonoplast in root and leaf. The coupling ratio of the H+ -ATPase also is a critical component. One proposed leak, that of Na+ influx across the plasma membrane through certain aquaporin channels, might be coupled to water flow, thus conserving energy. For the tonoplast, control of two types of cation channels is required for energy efficiency. Transporters controlling the Na+ and Cl- concentrations in mitochondria and chloroplasts are largely unknown and could be a major energy cost. The complexity of the system will require a sophisticated modelling approach to identify critical transporters, apoplastic barriers and root structures. This modelling approach will inform experimentation and allow a quantitative assessment of the energy costs of NaCl tolerance to guide breeding and engineering of molecular components.
Subject(s)
Crops, Agricultural/physiology , Energy Metabolism , Salt Tolerance/physiology , Biological Transport , Cell Respiration , Plant Roots/anatomy & histologyABSTRACT
Essential oil from the leaves of Artemisia vulgaris L. (Compositae) cultivated in Brazil was investigated for its chemical composition and biological activities including antibacterial, antifungal, and anthelmintic. The constituents of essential oils isolated by hydro-distillation were examined by GC-MS and a total of 18 components were identified. The essential oil was dominated by oxygenated sesquiterpenes (44.4%), sesquiterpene hydrocarbons (33.3%), and oxygenated monoterpenes (16.6%). Caryophyllene (37.45%), germacrene D (16.17%), and humulene (13.66%) were the major components. The essential oils from A. vulgaris showed bactericidal and fungicidal properties against Staphylococcus aureus and Candida albicans, respectively. Anthelmintic activity against Haemonchus contortus was absent in this essential oil. Altogether above results indicate that essential oils from A. vulgaris can be used for various medicinal purposes.
ABSTRACT
Oxidative stress signaling is essential for plant adaptation to hostile environments. Previous studies revealed the essentiality of hydroxyl radicals (HOâ¢)-induced activation of massive K⺠efflux and a smaller Ca2+ influx as an important component of plant adaptation to a broad range of abiotic stresses. Such activation would modify membrane potential making it more negative. Contrary to these expectations, here, we provide experimental evidence that HO⢠induces a strong depolarization, from -130 to -70 mV, which could only be explained by a substantial HOâ¢-induced efflux of intracellular anions. Application of Gd3+ and NPPB, non-specific blockers of cation and anion conductance, respectively, reduced HOâ¢-induced ion fluxes instantaneously, implying a direct block of the dual conductance. The selectivity of an early instantaneous HOâ¢-induced whole cell current fluctuated from more anionic to more cationic and vice versa, developing a higher cation selectivity at later times. The parallel electroneutral efflux of K⺠and anions should underlie a substantial leak of the cellular electrolyte, which may affect the cell's turgor and metabolic status. The physiological implications of these findings are discussed in the context of cell fate determination, and ROS and cytosolic K⺠signaling.
Subject(s)
Calcium/metabolism , Cell Membrane/metabolism , Hydroxyl Radical/metabolism , Oxidative Stress , Plant Roots/physiology , Potassium/metabolism , Adaptation, Physiological , Anions/chemistry , Anions/metabolism , Calcium/chemistry , Cell Membrane/chemistry , Hordeum/physiology , Hydroxyl Radical/chemistry , Ion Transport , Membrane Potentials , Patch-Clamp Techniques , Pisum sativum/physiology , Plant Roots/cytology , Potassium/chemistry , Protoplasts/ultrastructure , Reactive Oxygen Species/chemistry , Reactive Oxygen Species/metabolismABSTRACT
Salt stress impacts multiple aspects of plant metabolism and physiology. For instance it inhibits photosynthesis through stomatal limitation, causes excessive accumulation of sodium and chloride in chloroplasts, and disturbs chloroplast potassium homeostasis. Most research on salt stress has focused primarily on cytosolic ion homeostasis with few studies of how salt stress affects chloroplast ion homeostasis. This review asks the question whether membrane-transport processes and ionic relations are differentially regulated between glycophyte and halophyte chloroplasts and whether this contributes to the superior salt tolerance of halophytes. The available literature indicates that halophytes can overcome stomatal limitation by switching to CO2 concentrating mechanisms and increasing the number of chloroplasts per cell under saline conditions. Furthermore, salt entry into the chloroplast stroma may be critical for grana formation and photosystem II activity in halophytes but not in glycophytes. Salt also inhibits some stromal enzymes (e.g. fructose-1,6-bisphosphatase) to a lesser extent in halophyte species. Halophytes accumulate more chloride in chloroplasts than glycophytes and appear to use sodium in functional roles. We propose the molecular identities of candidate transporters that move sodium, chloride and potassium across chloroplast membranes and discuss how their operation may regulate photochemistry and photosystem I and II activity in chloroplasts.
Subject(s)
Chloroplasts/metabolism , Salt Tolerance , Salt-Tolerant Plants/physiology , Sodium Chloride/metabolism , Ion TransportABSTRACT
The aquaporin AtPIP2;1 is an abundant plasma membrane intrinsic protein in Arabidopsis thaliana that is implicated in stomatal closure, and is highly expressed in plasma membranes of root epidermal cells. When expressed in Xenopus laevis oocytes, AtPIP2;1 increased water permeability and induced a non-selective cation conductance mainly associated with Na+ . A mutation in the water pore, G103W, prevented both the ionic conductance and water permeability of PIP2;1. Co-expression of AtPIP2;1 with AtPIP1;2 increased water permeability but abolished the ionic conductance. AtPIP2;2 (93% identical to AtPIP2;1) similarly increased water permeability but not ionic conductance. The ionic conductance was inhibited by the application of extracellular Ca2+ and Cd2+ , with Ca2+ giving a biphasic dose-response with a prominent IC50 of 0.32 mм comparable with a previous report of Ca2+ sensitivity of a non-selective cation channel (NSCC) in Arabidopsis root protoplasts. Low external pH also inhibited ionic conductance (IC50 pH 6.8). Xenopus oocytes and Saccharomyces cerevisiae expressing AtPIP2;1 accumulated more Na+ than controls. Establishing whether AtPIP2;1 has dual ion and water permeability in planta will be important in understanding the roles of this aquaporin and if AtPIP2;1 is a candidate for a previously reported NSCC responsible for Ca2+ and pH sensitive Na+ entry into roots.
Subject(s)
Aquaporins/metabolism , Arabidopsis Proteins/metabolism , Calcium/metabolism , Amino Acid Substitution , Animals , Aquaporins/genetics , Arabidopsis Proteins/genetics , Cadmium/pharmacology , Calcium/pharmacology , Gene Expression Regulation, Plant , Glycine/genetics , Hydrogen-Ion Concentration , Oocytes/drug effects , Oocytes/physiology , Recombinant Proteins/genetics , Recombinant Proteins/metabolism , Saccharomyces cerevisiae/genetics , Sodium/metabolism , Tryptophan/genetics , Water/metabolism , Xenopus laevisABSTRACT
Soil salinity remains a major threat to global food security, and the progress in crop breeding for salinity stress tolerance may be achieved only by pyramiding key traits mediating plant adaptive responses to high amounts of dissolved salts in the rhizosphere. This task may be facilitated by studying natural variation in salinity tolerance among plant species and, specifically, exploring mechanisms of salinity tolerance in halophytes. The aim of this work was to establish the causal link between mesophyll ion transport activity and plant salt tolerance in a range of evolutionary contrasting halophyte and glycophyte species. Plants were grown under saline conditions in a glasshouse, followed by assessing their growth and photosynthetic performance. In a parallel set of experiments, net K+ and H+ transport across leaf mesophyll and their modulation by light were studied in control and salt-treated mesophyll segments using vibrating non-invasive ion selective microelectrode (the MIFE) technique. The reported results show that mesophyll cells in glycophyte species loses 2-6 fold more K+ compared with their halophyte counterparts. This decline was reflected in a reduced maximum photochemical efficiency of photosystem II, chlorophyll content and growth observed in the glasshouse experiments. In addition to reduced K+ efflux, the more tolerant species also exhibited reduced H+ efflux, which is interpreted as an energy-saving strategy allowing more resources to be redirected towards plant growth. It is concluded that the ability of mesophyll to retain K+ without a need to activate plasma membrane H+-ATPase is an essential component of salinity tolerance in halophytes and halophytic crop plants.
Subject(s)
Potassium/metabolism , Salt-Tolerant Plants/metabolism , Aizoaceae/metabolism , Beta vulgaris/metabolism , Chenopodium quinoa/metabolism , Chlorophyll/metabolism , Mesembryanthemum/metabolism , Mesophyll Cells/metabolism , Photosynthesis , Plant Leaves/metabolism , Salinity , Salt Tolerance/physiology , Species Specificity , Vicia faba/metabolismABSTRACT
While the importance of cell type specificity in plant adaptive responses is widely accepted, only a limited number of studies have addressed this issue at the functional level. We have combined electrophysiological, imaging, and biochemical techniques to reveal the physiological mechanisms conferring higher sensitivity of apical root cells to salinity in barley (Hordeum vulgare). We show that salinity application to the root apex arrests root growth in a highly tissue- and treatment-specific manner. Although salinity-induced transient net Na+ uptake was about 4-fold higher in the root apex compared with the mature zone, mature root cells accumulated more cytosolic and vacuolar Na+, suggesting that the higher sensitivity of apical cells to salt is not related to either enhanced Na+ exclusion or sequestration inside the root. Rather, the above differential sensitivity between the two zones originates from a 10-fold difference in K+ efflux between the mature zone and the apical region (much poorer in the root apex) of the root. Major factors contributing to this poor K+ retention ability are (1) an intrinsically lower H+-ATPase activity in the root apex, (2) greater salt-induced membrane depolarization, and (3) a higher reactive oxygen species production under NaCl and a larger density of reactive oxygen species-activated cation currents in the apex. Salinity treatment increased (2- to 5-fold) the content of 10 (out of 25 detected) amino acids in the root apex but not in the mature zone and changed the organic acid and sugar contents. The causal link between the observed changes in the root metabolic profile and the regulation of transporter activity is discussed.
Subject(s)
Acclimatization , Hordeum/enzymology , Hordeum/physiology , Plant Roots/enzymology , Potassium/metabolism , Proton-Translocating ATPases/metabolism , Salinity , Stress, Physiological , Acclimatization/drug effects , Allantoin/pharmacology , Cations/metabolism , Hordeum/drug effects , Metabolome/drug effects , Metabolomics , Models, Biological , Organ Specificity/drug effects , Plant Epidermis/cytology , Plant Epidermis/drug effects , Plant Epidermis/metabolism , Plant Roots/drug effects , Plant Roots/growth & development , Plant Roots/physiology , Reactive Oxygen Species/metabolism , Sodium/metabolism , Sodium Chloride/pharmacology , Stress, Physiological/drug effectsABSTRACT
Brassica species are known to possess significant inter and intraspecies variability in salinity stress tolerance, but the cell-specific mechanisms conferring this difference remain elusive. In this work, the role and relative contribution of several key plasma membrane transporters to salinity stress tolerance were evaluated in three Brassica species (B. napus, B. juncea, and B. oleracea) using a range of electrophysiological assays. Initial root growth assay and viability staining revealed that B. napus was most tolerant amongst the three species, followed by B. juncea and B. oleracea At the mechanistic level, this difference was conferred by at least three complementary physiological mechanisms: (i) higher Na(+) extrusion ability from roots resulting from increased expression and activity of plasma membrane SOS1-like Na(+)/H(+) exchangers; (ii) better root K(+) retention ability resulting from stress-inducible activation of H(+)-ATPase and ability to maintain more negative membrane potential under saline conditions; and (iii) reduced sensitivity of B. napus root K(+)-permeable channels to reactive oxygen species (ROS). The last two mechanisms played the dominant role and conferred most of the differential salt sensitivity between species. Brassica napus plants were also more efficient in preventing the stress-induced increase in GORK transcript levels and up-regulation of expression of AKT1, HAK5, and HKT1 transporter genes. Taken together, our data provide the mechanistic explanation for differential salt stress sensitivity amongst these species and shed light on transcriptional and post-translational regulation of key ion transport systems involved in the maintenance of the root plasma membrane potential and cytosolic K/Na ratio as a key attribute for salt tolerance in Brassica species.
Subject(s)
Brassica napus/physiology , Brassica/physiology , Mustard Plant/physiology , Plant Roots/physiology , Potassium Channels/physiology , Potassium/metabolism , Salt Tolerance/physiology , Brassica/metabolism , Brassica napus/metabolism , Calcium-Transporting ATPases/metabolism , Calcium-Transporting ATPases/physiology , Gene Expression Regulation, Plant/physiology , Membrane Potentials/physiology , Mustard Plant/metabolism , Plant Roots/metabolism , Potassium Channels/metabolism , Reactive Oxygen Species/metabolismABSTRACT
Three different species of Brassica, with differential salt sensitivity were used to understand physiological mechanisms of salt tolerance operating in these species and to evaluate the relative contribution of different strategies to cope with salt load. Brassica napus was the most tolerant species in terms of the overall performance, with Brassica juncea and Brassica oleracea being much more sensitive to salt stress with no obvious difference between them. While prominent reduction in net CO2 assimilation was observed in both sensitive species, physiological mechanisms beyond this reduction differed strongly. Brassica juncea plants possessed high osmotolerance and were able to maintain high transpiration rate but showed a significant reduction in leaf chlorophyll content and efficiency of leaf photochemistry. On the contrary, B. oleracea plants possessed the highest (among the three species) tissue tolerance but showed a very significant stomatal limitation of photosynthesis. Electrophysiological experiments revealed that the high tissue tolerance in B. oleracea was related to the ability of leaf mesophyll cells to maintain highly negative membrane potential in the presence of high apoplastic Na(+) . In addition to high osmotolerance, the most tolerant B. napus showed also lesser accumulation of toxic Na(+) and Cl(-) in the leaf, possessed moderate tissue tolerance and had a superior K(+) retention ability. Taken together, the results from this study indicate that the three Brassica species employ very different mechanisms to cope with salinity and, despite its overall sensitivity to salinity, B. oleracea could be recommended as a valuable 'donor' of tissue tolerance genes to confer this trait for marker-assisted breeding programs.
Subject(s)
Brassica/physiology , Plant Transpiration , Sodium Chloride/pharmacology , Brassica/drug effects , Brassica/radiation effects , Chlorophyll/metabolism , Light , Osmotic Pressure , Photochemistry , Photosynthesis , Plant Leaves/drug effects , Plant Leaves/physiology , Plant Leaves/radiation effects , Plant Stomata/drug effects , Plant Stomata/physiology , Plant Stomata/radiation effects , Salinity , Salt Tolerance , Sodium Chloride/metabolism , Species Specificity , Stress, PhysiologicalABSTRACT
Abiotic stresses such as salinity, drought, and flooding severely limit food and fibre production and result in penalties of in excess of US$100 billion per annum to the agricultural sector. Improved abiotic stress tolerance to these environmental constraints via traditional or molecular breeding practices requires a good understanding of the physiological and molecular mechanisms behind roots sensing of hostile soils, as well as downstream signalling cascades to effectors mediating plant adaptive responses to the environment. In this review, we discuss some common mechanisms conferring plant tolerance to these three major abiotic stresses. Central to our discussion are: (i) the essentiality of membrane potential maintenance and ATP production/availability and its use for metabolic versus adaptive responses; (ii) reactive oxygen species and Ca(2+) 'signatures' mediating stress signalling; and (iii) cytosolic K(+) as the common denominator of plant adaptive responses. We discuss in detail how key plasma membrane and tonoplast transporters are regulated by various signalling molecules and processes observed in plants under stress conditions (e.g. changes in membrane potential; cytosolic pH and Ca(2+); reactive oxygen species; polyamines; abscisic acid) and how these stress-induced changes are related to expression and activity of specific ion transporters. The reported results are then discussed in the context of strategies for breeding crops with improved abiotic stress tolerance. We also discuss a classical trade-off between tolerance and yield, and possible avenues for resolving this dilemma.