Physiological and molecular characterization of salt response of <Emphasis Type="Italic">Arabidopsis thaliana NOK2</Emphasis> ecotype

Arabidopsis thaliana is a glycophyte capable to tolerate mild salinity. Although salt sensitivity of this species, a variability of this characteristic was revealed between different ecotypes. This study presents the physiological and molecular characteristics of salt response of two ecotypes, NOK2 and Columbia (Col). Seedlings were cultivated in hydroponics in the presence of 0 or 50 mM NaCl during 25 days. Rosette leaf samples were collected after 19, 22, and 25 days for determination of physiological parameters, and after 18 days for study of DNA polymorphism. Salt treatment decreased rosette dry matter, leaf number, leaf hydration, and leaf surface area. All these effects were significantly more visible in Col than in NOK2. Moreover, the NOK2 leaves accumulated less Na⁺ and more K⁺ than those of Col. DNA polymorphism between the two ecotypes was analyzed with codominant molecular markers based on PCR amplification, namely, microsatellites, cleaved amplified polymorphism sequence (CAPS), and single nucleotide polymorphism markers (SNP). Among the 35 tested markers, 17 showed a clear polymorphism and were distributed on the five Arabidopsis chromosomes ending with a genetic map construction. These results could play an important role in the future establishment of cartography of candidate gene controlling the K⁺/Na⁺ selectivity of ion transport in leaves, a component of plant salt tolerance.     

Responses of some genetically diverse lines of chick pea (Cicer arietinum L.) to salt

The salt tolerance of three tolerant accessions of chick pea, CM 663, 10130 and 10572 and three sensitive accessions 10582, 12908 and 12909 selected at the germination and seedling stage was assessed at the adult stage using sand culture salinized with 0, 40 or 80 mol m^-3 NaCl. The two tolerant accessions, CM 663 and 10572 and one sensitive, 12908 showed consistent correlation between the degrees of salt tolerance at the early growth stages and adult stage as the former two produced significantly higher seed yield compared with the other accessions and the latter did not survive till seed setting in the salt treatments. By contrast 10130 which was found relatively salt tolerant at the two early growth stages could not survive in 40 mol m^-3 NaCl till seed setting. Similarly two sensitive accessions, 10582 and 12909 not only survived at the adult stage but produced some yield as well. On the basis of performance of the six accessions at three different stages, accessions CM 663 and 10572 can be categorised as relatively salt tolerant, 12908 as sensitive and 10130, 10582 and 12909 as moderately tolerant. The tolerant accession CM 663 had high Na⁺ and Cl^- in the leaves but maintained high K:Na ratios and high K⁺ versus Na⁺ selectivity. This accession had relatively low leaf osmotic potential which may be due to its high accumulation of Na⁺ and Cl^- in the leaves. By contrast the second tolerant accession 10572 had lowest Na⁺ and moderate Cl^- in the leaves.of all accessions but had highest K⁺ versus Na⁺ selectivity, although its leaf K:Na was intermediate. It had also relatively low osmotic potential which cannot be related to different ions determined in this study. The salt sensitive accession 12908 had high leaf Na⁺ and moderate Cl^- but had very low K:Na ratio (less than one) and K⁺ versus Na⁺ selectivity. The remaining accessions as a whole did not show any consistent pattern of uptake of different ions. The positive correlation between the degree of salt tolerance at different growth stages do exist in some accessions of chick pea examined in the present study, but for others in which no positive correlation was observed suggests that a combination of certain characters can be used as selection criterion for improving salt tolerance in chick pea.     

Photosynthesis and ion content of leaves and isolated chloroplasts of salt-stressed spinach

Spinach (Spinacia oleracea) plants were subjected to salt stress by adding NaCl to the nutrient solution in increments of 25 millimolar per day to a final concentration of 200 millimolar. Plants were harvested 3 weeks after starting NaCl treatment. Fresh and dry weight of both shoots and roots was decreased more than 50% compared to control plants but the salt-stressed plants appeared healthy and were still actively growing. The salt-stressed plants had much thicker leaves. The salt-treated plants osmotically adjusted to maintain leaf turgor. Leaf K⁺ was decreased but Na⁺ and Cl⁻ were greatly increased.

The potential photosynthetic capacity of the leaves was measured at saturating CO₂ to overcome any stomatal limitation. Photosynthesis of salt-stressed plants varied only by about 10% from the controls when expressed on a leaf area or chlorophyll basis. The yield of variable chlorophyll a fluorescence from leaves was not affected by salt stress. Stomatal conductance decreased 70% in response to salt treatment.

Uncoupled rates of electron transport by isolated intact chloroplasts and by thylakoids were only 10 to 20% below those for control plants. CO₂-dependent O₂ evolution was decreased by 20% in chloroplasts isolated from salt-stressed plants. The concentration of K⁺ in the chloroplast decreased by 50% in the salt-stressed plants, Na⁺ increased by 70%, and Cl⁻ increased by less than 20% despite large increases in leaf Na⁺ and Cl⁻.

It is concluded that, for spinach, salt stress does not result in any major decrease in the photosynthetic potential of the leaf. Actual photosynthesis by the plant may be reduced by other factors such as decreased stomatal conductance and decreased leaf area. Effective compartmentation of ions within the cell may prevent the accumulation of inhibitory levels of Na⁺ and Cl⁻ in the chloroplast.

     

The mechanism of stomatal closing by salicylic acid inCommelina communis L.

The mechanism of stomatal closing by salicylic acid (SA) has been investigated. The addition of 1 mM SA to fully opened stomata resulted in a significant reduction of 75% in stomatal aperture. Stomata in the treatment of SA with EGTA closed as observed in the treatment of SA. However, the addition of catalase with SA completely inhibited stomatal closing. Stomatal closing induced by SA was also reduced by Ca²⁺. To understand the relation bewteen stomatal closing by SA and catalase activity, the effect of SA on catalse activity and the effect of AT (catalase inhibitor) on stomatal closing was investigated. SA inhibited 32% of catalase activity. Stomata in isolated epidermis floated on an incubation solution containing 0.1 mM AT closed from 9.6 μm to 3.2 μm after 1 hour. SA stimulated K⁺ efflux as much as the twice of the control in isolated strips. SA inhibited 53% of photosynthetic activity at the light intensity of 1000 μmole m² s¹ on SA infiltrated leaves. A similar result was found on stomatal conductance in SA infiltrated leaves. These results indicate that SA inhibit catalase activity and increase the concentration of H₂O₂ in guard cell cytoplasm. H₂O₂ oxidize the plasma membrane and increase the membrane permeability of K⁺. The mass efflux of K⁺ induce the loss of turgor pressure and lead to stomatal closing. The inhibition of photosynthetic activity by SA suggests that stomatal closing by SA is also related with the decrease of photosynthetic activity.     

Effect of sodium chloride on gas exchange, antioxidative defense mechanism and ion accumulation in different cultivars of Indian jujube (Ziziphus mauritiana L.)

An experiment was conducted to study the effect of NaCl (electric conductivity of 0, 4, 8, 12, and 16 dS m^?1) on growth, gas exchange parameters, water status, membrane injury, chlorophyll stability index and oxidative defense mechanisms in two cultivars (Gola and Umran) of Indian jujube (Ziziphus mauritiana). Results showed that the dry mass and leaf area reduced linearly with increasing levels of salinity. Net photosynthetic rate (P _N), transpiration (E), and stomatal conductance (g _s) were comparatively lower in Umran which further declined with salinity. Leaf relative water content, chlorophyll (Chl) stability and membrane stability also decreased significantly under salt stress, with higher magnitude in Umran. Superoxide dismutase (SOD), peroxidase (POX) and catalase (CAT) activities were higher in Gola whereas hydrogen peroxide (H₂O₂) accumulation and lipid peroxidation (MDA content) were higher in control as well as salttreated plants of Umran. The Na⁺ content was higher in the roots of Gola and in the leaves of Umran, resulting in high K⁺/Na⁺ ratio in Gola leaves. Thus it is suggested that salt tolerance mechanism is more efficiently operative in cultivar Gola owing to better management of growth, physiological attributes, antioxidative defense mechanism, and restricted translocation of Na⁺ from root to leaves along with larger accumulation of K⁺ in its leaves.     

Effects of NaCl stress on the growth and photosynthetic characteristics of Ulmus pumila L. seedlings in sand culture

The effects of NaCl stress on the growth and photosynthetic characters of Ulmus pumila L. seedlings were investigated under sand culture condition. With increasing NaCl concentration, main stem height, branch number, leaf number, and leaf area declined, while Na⁺ content and the Na⁺/K⁺ ratio in both expanded and expanding leaves increased. Na⁺ content was significantly higher in expanded leaves than in those just expanding. Chlorophyll (Chl) a and Chl b contents declined as NaCl concentration increased. The net photosynthetic rate, intercellular CO₂ concentration, stomatal conductance, and transpiration rate also declined, but stomatal limitation value increased as NaCl concentration increased. Both the maximal quantum yield of PSII photochemistry and the effective quantum yield of PSII photochemistry declined as NaCl concentration rose. These results suggest that the accumulation of Na⁺ in already expanded leaves might reduce damage to the expanding leaves and help U. pumila endure high salinity. The reduced photosynthesis in response to salt stress was mainly caused by stomatal limitation.     

Analysis of salinity effects on basil leaf surface area, photosynthetic activity, and growth

Basil (Ocimum basilicum L.) seedlings were cultured on liquid medium in controlled conditions. Two varieties differing in leaf size were compared. When plants were 30?days old, the medium was supplemented with 50?mM NaCl. After 15?days of treatment, root, stem and leaf biomass, leaf number, and leaf surface area were measured. Ion accumulation was determined in roots, stems, and leaves. Photosynthetic parameters (CO₂ fixation rate, internal CO₂ concentration, stomatal conductance) as well as transpiration rate were determined on separate leaves. Electrolyte leakage and malondialdehyde content were used to estimate damage to membranes and lipid peroxidation, respectively. Several antioxidant enzymatic activities were used as proxies of oxidative stress. High Na⁺ concentration was reached in leaf tissues. Salt restricted whole plant biomass deposition rate by diminishing leaf number and leaf expansion, as well as photosynthetic activity were estimated from whole plant biomass production per unit leaf surface area. Diminished stomatal conductance restricted CO₂ fixation rate, and decrease in chlorophyll content presumably limited photosynthetic activity. Lipid peroxidation revealed damages to membranes. The magnitude of these responses differed between the two varieties, indicating that an intraspecific variability in salt response exists in basil.     

Exogenous proline effects on water relations and ions contents in leaves and roots of young olive

The ability of exogenous compatible solutes, such as proline, to counteract salt inhibitory effects was investigated in 2-year-old olive trees (Olea europaea L. cv. Chemlali) subjected to different saline water irrigation levels supplied or not with exogenous proline. Leaf water relations [relative water content (RWC), water potential], photosynthetic activity, leaf chlorophyll content, and starch contents were measured in young and old leaves. Salt ions (Na⁺, K⁺, and Ca²⁺), proline and soluble sugars contents were determined in leaf and root tissues. Supplementary proline significantly mitigated the adverse effects of salinity via the improvement of photosynthetic activity (Pn), RWC, chlorophyll and carotenoid, and starch contents. Pn of young leaves in the presence of 25 mM proline was at 1.18 and 1.38 times higher than the values recorded under moderate (SS1) and high salinity (SS2) treatments, respectively. Further, the proline supply seems to have a more important relaxing effect on the photosynthetic chain in young than in old leaves of salt-stressed olive plants. The differential pattern of proline content between young and old leaves suggests that there would be a difference between these tissues in distinguishing between the proline taken from the growing media and that produced as a result of salinity stress. Besides, the large reduction in Na⁺ accumulation in leaves and roots in the presence of proline could be due to its interference in osmotic adjustment process and/or its dilution by proline supply. Moreover, the lower accumulation of Na⁺ in proline-treated plants, compared to their corresponding salinity treatment, displayed the improved effect of proline on the ability of roots to exclude the salt ions from the xylem sap flowing to the shoot, and thus better growth rates.     

Comprehensive evaluating of wild and cultivated emmer wheat (<Emphasis Type="Italic">Triticum turgidum</Emphasis> L.) genotypes response to salt stress

Emmer wheat as the progenitor of common wheat, holds the genetic potentiality for improvement of wheat yield, quality and stress tolerance such as drought and salt. To comprehensively evaluate the salt tolerance of emmer wheat, a total of 30 traits including growth, physiology and photosynthesis related as well as K⁺ and Na⁺ content of 30 wild emmer and 14 durum wheat accessions were systematically investigated and compared between normal and saline conditions. Salt tolerance index (STI) based on multiple regression analysis of these traits was calculated and five wild emmer accessions showed high salt tolerance, which could be used as valuable resource for wheat salt tolerance improvement. Furthermore, wild emmer genotypes showed wider trait performance variation compared to durum wheat, indicating the higher genetic diversity in wild emmer wheat. Then, shoot Na⁺ content, shoot K⁺/Na⁺ ratio, root length and root surface area were identified as suitable indexes for salt tolerance evaluation. Na⁺ exclusion mechanism was found to be playing an important role in response to salt stress in emmer wheat. The salt tolerance in emmer wheat was systematically characterized here, which not only provided the elite germplasm for wheat improvement, but also provided the efficient method and some useful indexes for salt tolerance assessing.     

Effect of NaCl on leaf salt secretion and antioxidative enzyme level in roots of a mangrove, Aegiceras corniculatum

Short-term salt (NaCl) treatment on Aegiceras corniculatum in roots and leaves showed no change in fresh and dry weight of leaves, roots and leaf area. There was no significant change in total soluble root protein, photosynthetic pigments of leaves and spectral characteristics of thylakoids. However, the activity of antioxidative enzymes (catalase, ascorbate peroxidase and guaiacol peroxidase) in roots decreased by 72, 58 and 80% respectively after 96 hr of treatment (300 mM of NaCl). Secretion of salts from the leaf salt glands and salt accumulation on upper surface of the leaves were quantified that revealed linear increase of salt secretion of leaf with increase in period of salt treatment. It was concluded that loss of activities of antioxidative enzymes at high salt treatment, caused leaf senescence in spite of high rates of salt secretion by Aegiceras corniculatum.     

Assessment of a non-destructive method to estimate the leaf area of <Emphasis Type="Italic">Armoracia rusticana</Emphasis>

The aim of the study was to analyze horseradish growth for developing a mathematical model to estimate the leaf area based on linear measurements of the leaf surface. Leaf area (LA), number, and morphometric characteristics of the leaves including lamina length (L) and width (W) were evaluated on two horseradish accessions (Cor and Mon) throughout a 2 year growing cycle. In both accessions, increased values of LA and leaf number were found by comparing the second with the first-growing season. Leaf development occurs along with variations in size and not in shape during the plant growth. The leaves are elliptical in shape but tend to be wider and bigger in Cor accession and tapered and similar to narrow ellipses in Mon showing different length/width relationship. Consequently, several regression models relating to the LA and L, W, L², and W² individually or in combination were fitted for each accession based on a set of 1000 leaves. The horseradish LA can be predicted based on either length or width alone. However, the regression linear model LA?=?aLW?+?b (LA?=?0.71LW ??0.27 and LA?=?0.76LW ??3.22 for Cor and Mon, respectively) provided the best LA estimation (R²?>?0.95). The validation of this latter model showed high correlation between LA measured and LA predicted in both accessions (R²?=?0.98). Considering the type of foliage of horseradish, the proposed model can be used to estimate the leaf area throughout the entire crop cycle.     

Responses of Arabidopsis thaliana to bicarbonate-induced iron deficiency

Morpho-physiological and biochemical responses of Arabidopsis thaliana (accession N1438) to bicarbonate-induced iron deficiency were investigated. Plants were grown in cabinet under controlled conditions, in a nutrient solution containing 5 μM Fe, added or not with 10 mM NaHCO₃. After 30 days, bicarbonate-treated plants displayed significantly lower biomass, leaf number and leaf surface area as compared to control plants, and slight yellowing of their younger leaves was observed. Potassium (K⁺) content was not modified by bicarbonate treatment in roots, whereas it was significantly diminished in shoots. Their content in ferrous iron (Fe²⁺) and in leaf total chlorophylls was noticeably lower than in control plants. Root Fe(III)-chelate reductase and phosphoenolpyruvate carboxylase (PEPC) activities were significantly enhanced, but leaf ribulose 1.5-bisphosphate carboxylase (Rubisco) activity was decreased.     

盐胁迫对3种平欧杂种榛幼苗叶片解剖结构及离子吸收、运输与分配的影响

研究盐胁迫下3个品种平欧杂种榛幼苗叶片解剖结构和离子代谢特征,以揭示盐胁迫响应与适应机制及不同品种的耐盐性差异。以‘达维’、‘辽榛7号’、‘玉坠’2年生压条苗为材料,在盆栽条件下经轻度、中度、重度(分别为50、100、200 mmol/L NaCl)盐胁迫处理,设对照为0,研究幼苗叶片显微解剖结构参数和Na~+、K~+、Cl~-、Ca²⁺含量的变化及其在根、茎、叶中的吸收、运输和分配特征。不同品种平欧杂种榛叶片厚度、上表皮厚度、下表皮厚度、栅栏组织和海绵组织厚度随着盐胁迫程度的增强呈现出先增加后降低的特点,轻度和中度胁迫下各参数显著高于对照。中度盐胁迫显著提高了各品种叶片结构紧密度。盐胁迫导致平欧杂种榛根、茎、叶Na~+和Cl~-含量明显高于对照。盐胁迫下,Na~+和Cl~-在叶中的绝对含量明显高于茎和根,但二者的增幅以根中最大,叶中最小,表明平欧杂种榛根系首先会吸收并截留一定数量的Na~+和Cl~-,然后将其运输至茎和叶中。与对照相比,轻度和中度盐胁迫下根、茎对K~+和Ca²⁺的吸收保持稳定或减少,叶对K~+和Ca²⁺...     

The strategy of Na+ compartmentation and growth of Atriplex centralasiatica in adaptation to saline environments

In this study, we investigated the adaptation strategy employed by Atriplex centralasiatica Iljin in response to high salinity. When grown in high saline environments (100–200 mM NaCl), A. centralasiatica plants were larger and more succulent. This increased growth and water uptake was correlated with a large and specific cellular accumulation of sodium, demonstrating that in A. centralasiatica Na⁺ is beneficial rather than toxic. More than 95% of Na⁺ absorbed by salt-treated A. centralasiatica plants accumulated in shoots, especially in leaves; approximately 98% of Na⁺ that accumulated in leaves was localized in leaf protoplasts, a situation that was responsible for the decreased photosynthetic rate observed with increasing salt concentration. Because of the greater leaf area per plant found under saline conditions, no reduction in biomass of individual plants was observed. Measurements on isolated tonoplast-enriched membrane vesicles derived from the leaves of A. centralasiatica revealed increased V-H⁺-ATPase hydrolytic activity and V-H⁺-ATPase proton pump activity in salt-treated leaves compared with controls. These results suggest that, as an adaptation to saline environments, A. centralasiatica can efficiently sequester Na⁺ into vacuoles, thereby increasing leaf area to maintain its CO₂ assimilation capabilities.     

Effects of calcium deficiency on Coffea arabica. Nutrient changes and correlation of calcium levels with some photosynthetic parameters

Calcium deficiency was induced in hydroponically grown 1.5-years-old coffee plants with 12–14 pairs of leaves. Calcium was given in the form of Ca(NO₃)₂: 5, 2.5, 0.1, 0.01 and 0 mM. After 71 days of Ca-treatment root and shoot as well as total biomass were decreased by severe Ca-deficiency. However, a stronger decrease was observed for shoot growth as revealed by the increase in the root/shoot ratio. New leaves were affected showing decreases in the total leaf area and in Leaf Area Duration (LAD). After 91 days of deficiency, leaf protein concentration decreased (by about 45%) in the top leaves while nitrate reductase activity (NRA) and NO₃ content showed no significant changes. Total nitrogen and mineral concentrations (P, K, Ca, Mg and Na) were also determined in leaves and roots. With the decrease in calcium concentration in Ca-deficiency conditions, we observed concomitant increases in the concentrations of K⁺, Mg²⁺ and Na⁺ in leaves (maximal changes of 32% for K⁺, 96% for Mg²⁺ and 438% for Na⁺) and in roots (108% for K⁺, 86% for Mg²⁺ and 38% for Na⁺). Accordingly, the ratio between elements changed, including the ratio N/P, showing a non-equilibrium in the balance of nutrients. Significant correlations were obtained between Ca²⁺ concentration and some photosynthetic parameters. Ca-deficiency conditions would increase the loss of energy as expressed by the rise in a_E and decrease the photochemical efficiency, which confirms the importance of this element in the stabilization of chlorophyll and in the maintenance of good photochemical efficiency at PS II level.Abbreviations Chl Chlorophyll - Fv/Fm ratio of variable to maximal fluorescence - LAD leaf area duration - LHC II light harvesting complex of PS II - NRA nitrate reductase activity - PC photosynthetic capacity - PS II photosystem II - P₆₈₀ reaction center of PS II - q_N non-photochemical quenching - q_E high-energy dependent quenching - q_p photochemical quenching - SLA specific leaf area     

Effects of 24‐epibrassinolide on plant growth,osmotic regulation and ion homeostasis of salt‐stressed canola

This study evaluated effects of foliar spraying 24‐epibrassinoide (24‐EBL) on the growth of salt‐stressed canola. Seedlings at the four‐leaf stage were treated with 150 mm NaCl and different concentrations of 24‐EBL (10^?6, 10^?8, 10^?10, 10^?12 m ) for 15 days. A concentration of 10^?10 m 24‐EBL was chosen as optimal and used in a subsequent experiment on plant biomass and leaf water potential parameters. The results showed that 24‐EBL mainly promoted shoot growth of salt‐stressed plants and also ameliorated leaf water status. Foliar spraying of salt‐stressed canola with 24‐EBL increased osmotic adjustment ability in all organs, especially in younger leaves and roots. This was mainly due to an increase of free amino acid content in upper leaves, soluble sugars in middle leaves, organic acids and proline in lower leaves, all of these compounds in roots, as well as essential inorganic ions. Na⁺ and Cl^? sharply increased in different organs under salt stress, and 24‐EBL reduced their accumulation. 24‐EBL improved the uptake of K⁺, Ca²⁺, Mg²⁺ and NO₃^? in roots, which were mainly transported to upper leaves, while NO₃^? was mainly transported to middle leaves. Thus, 24‐EBL improvements in ion homeostasis of K⁺/Na⁺, Ca²⁺/Na⁺, Mg²⁺/Na⁺ and NO₃^?/Cl^?, especially in younger leaves and roots, could be explained. As most important parts, younger leaves and roots were the main organs protected by 24‐EBL via improvement in osmotic adjustment ability and ion homeostasis. Further, physiological status of growth of salt‐stressed canola was ameliorated after 24‐EBL treatment.     

Effect of potassium on growth and nitrate reductase during water stress and recovery in maize

Abstract The effect of potassium (0,50, 100 and 200 mg/pot) was studied on growth characteristics and nitrate reductase activity in maize (Zea mays) seedlings during water stress and subsequent recovery. In irrigated plants K⁺ increased the rate of leaf area expansion, leading to increased leaf area per plant. Increased leaf area was associated with decreased chlorophyll content. Water stress (–15 bars) enhanced the stomatal resistance of leaves which was further accentuated by K⁺ application. Nitrate reductase activity rose in irrigated plants 24 h after K⁺ application. Subsequently, as water stress developed, K⁺ helped to maintain higher NR activity for the first two days. However, K⁺ had no effect on half life of NR in light or darkness. During recovery from stress K⁺ aided to maintain the higher leaf expansion rate, the chlorophyll content and the stomatal resistance. The results above are discussed in relation to the ability of K⁺ to maintain better growth under water stress.