Genetic control and mechanisms of salt and hyperosmotic shock resistance in cyanobacteria

Exposure to high concentrations of environmental NaCl exerts two stress effects on living cells, increasing the osmotic pressure and the concentration of inorganic ions. Salt stress dramatically suppresses the photosynthetic activity in cells of phototrophic organisms, such as cyanobacteria. During salt adaptation, cyanobacterial cells accumulate osmoprotectors, export excessive Na+ with the help of Na+/H+ antiporters, and actively absorb K+ with the help of K+-transporting systems. These physiological processes are accompanied by induction or suppression of several genes involved in salt adaptation. The review considers the main mechanisms responsible for the resistance of cyanobacterial cells to salt and hyperosmotic stresses. Special emphasis is placed on recent achievements in studying the genetic control of salt resistance and regulation of gene expression during adaptation of cyanobacteria to salt and hyperosmotic stresses.     

[14C]Carbon-dioxide fixation by isolated leaf epidermes with stomata closed or open

Following small hypo-osmotic shocks, ion concentrations (Na⁺, K⁺, Cl^-) in Platymonas subcordiformis decreased; this was due mainly to an increase of cell volume. With larger hypo-osmotic stresses, the decrease of ion concentration continued and, additionally, extrusion of mannitol was observed. The ion and mannitol concentrations were not regained after 240 min. In contrast, following hyperosmotic shocks, the ion concentrations increased transitorily during the first 20–40 min. The same was true for K⁺ following small hyperosmotic stresses and for Na⁺ and — partially — Cl^- with larger shocks. Large hyperosmotic stresses caused permanent accumulation of mannitol, which levelled off after 60–80 min. Thus the transient increase of ions bridged the concentration gap until mannitol was accumulated to a high enough concentration to account for the osmotic adaptation of Platymonas, together with a basal level of the ions K⁺, Na⁺, Cl^-.Abbreviations PS photosynthesis - Resp respiration     

Osmotic adaptation in halotolerant yeast, Debaryomyces nepalensis NCYC 3413: role of osmolytes and cation transport

Debaryomyces nepalensis NCYC 3413, a food spoiling yeast isolated from rotten apple, has been previously demonstrated as halotolerant yeast. In the present study, we assessed its growth, change in cell size, and measured the intracellular polyol and cations (Na⁺ or K⁺) accumulated during growth in the absence and presence of different concentrations of salts (NaCl and KCl). Cells could tolerate 2 M NaCl and KCl in defined medium. Scanning electron microscopic results showed linear decrease in mean cell diameter with increase in medium salinity. Cells accumulated high amounts of K⁺ during growth at high concentrations of KCl. However, it accumulated low amounts of Na⁺ and high amounts of K⁺ when grown in the presence of NaCl. Cells grown in the absence of salt showed rapid influx of Na⁺/K⁺ on incubation with high salt. On incubation with 2 M KCl, cells grown at 2 M NaCl showed an immediate efflux of Na⁺ and rapid uptake of K⁺ and vice versa. To withstand the salt stress, osmotic adjustment of intracellular cation was accompanied by intracellular accumulation of polyol (glycerol, arabitol, and sorbitol). Based on our result, we hypothesize that there exists a balanced efflux and synthesis of osmolytes when D. nepalensis was exposed to hypoosmotic and hyperosmotic stress conditions, respectively. Our findings suggest that D. nepalensis is an Na⁺ excluder yeast and it has an efficient transport system for sodium extrusion.     

Na<Superscript>+</Superscript>/H<Superscript>+</Superscript> and K<Superscript>+</Superscript>/H<Superscript>+</Superscript> antiporters AtNHX1 and AtNHX3 from <Emphasis Type="Italic">Arabidopsis</Emphasis> improve salt and drought tolerance in transgenic poplar

The tonoplast and plasma membrane localized sodium (potassium)/proton antiporters have been shown to play an important role in plant resistance to salt stress. In this study, AtNHX1 and AtNHX3, two tonoplast Na⁺(K⁺)/H⁺ antiporter encoding genes from Arabidopsis thaliana, were expressed in poplar to investigate their biological functions in the resistance to abiotic stresses in woody plants. Transgenic poplar plants expressing either gene exhibited increased resistance to both salt and water-deficit stresses. Compared to the wild type (WT) plants, transgenic plants accumulated more sodium and potassium ions in the presence of 100 mM NaCl and showed reduced electrolyte leakage in the leaves under water stress. Furthermore, the proton-translocating and cation-dependent H⁺ (Na⁺/H⁺ or K⁺/H⁺) exchange activities in the tonoplast vesicles isolated from the leaves of transgenic plants were higher than in those isolated from WT plants. Therefore, constitutive expression of either AtNHX1 or AtNHX3 genetically modified the salt and water stress tolerance of transgenic poplar plants, providing a potential tool for engineering tree species with enhanced resistance to multiple abitotic stresses.     

Relationship between Sodium Influx and Salt Tolerance of Nitrogen-Fixing Cyanobacteria

The relationship between sodium uptake and cyanobacterial salt (NaCl) tolerance has been examined in two filamentous, heterocystous, nitrogen-fixing species of Anabaena. During diazotrophic growth at neutral pH of the growth medium, Anabaena sp. strain L-31, a freshwater strain, showed threefold higher uptake of Na⁺ than Anabaena torulosa, a brackish-water strain, and was considerably less salt tolerant (50% lethal dose of NaCl, 55 mM) than the latter (50% lethal dose of NaCl, 170 mM). Alkaline pH or excess K⁺ (>25 mM) in the medium causes membrane depolarization and inhibits Na⁺ influx in both cyanobacteria (S. K. Apte and J. Thomas, Eur. J. Biochem. 154:395-401, 1986). The presence of nitrate or ammonium in the medium caused inhibition of Na⁺ influx accompanied by membrane depolarization. These experimental manipulations affecting Na⁺ uptake demonstrated a good negative correlation between Na⁺ influx and salt tolerance. All treatments which inhibited Na⁺ influx (such as alkaline pH, K⁺ above 25 mM, NO₃⁻, and NH₄⁺), enhanced salt tolerance of not only the brackish-water but also the freshwater cyanobacterium. The results indicate that curtailment of Na⁺ influx, whether inherent or effected by certain environmental factors (e.g., combined nitrogen, alkaline pH), is a major mechanism of salt tolerance in cyanobacteria.     

盐旱交叉胁迫对柽柳幼苗渗透调节物质含量的影响

以2年生柽柳幼苗为试验材料,采用盆栽模拟试验研究不同盐分和干旱胁迫对其叶片中渗透调节物质的影响,以探讨柽柳幼苗对盐旱交叉胁迫的适应性.结果表明:(1)随盐旱胁迫的不断加剧,幼苗叶片中可溶性糖含量呈先升高后降低的趋势,且中度和重度盐旱胁迫下均显著高于对照(CK).(2)幼苗叶片中脯氨酸含量在不同盐旱胁迫下均呈逐渐上升趋势,但在重度盐分和中度、重度干旱交叉胁迫下显著高于CK.(3)幼苗叶片中Na+、Cl含量在不同干旱胁迫下,随盐胁迫的加剧呈不同的变化规律,盐旱胁迫的各个处理水平下均显著高于CK,而K+、Ca2+、SO42-含量在轻度和重度干旱胁迫下随盐胁迫增强不断降低.(4)在中度盐旱胁迫下,K+、Ca2+含量与CK无明显差异.研究表明,柽柳幼苗中渗透调节物质在其抗旱耐盐性方面具有积极的调节作用;柽柳幼苗在盐旱胁迫下表现出一定的交叉适应性,适度的干旱胁迫能增强柽柳幼苗对盐分胁迫的耐受能力.     

不同倍性小麦对盐胁迫的适应性差异

植物染料在工业化应用过程中存在着资源限制,目标色相不丰富、色牢度不理想、植物染料本身的鉴别和成品的鉴别等问题。为了丰富染料植物资源的来源和提高染料植物资源的利用效率,该研究对西双版纳傣族利用的染料植物及其染色工艺涉及的相关植物进行了系统调查。2014年10月到2016年1月,采用半结构式访谈法对西双版纳14个村寨的56个关键信息人进行访谈,收集信息包括使用着色植物、媒染植物和助染植物的种类、傣名、利用部位和资源来历,以及预处理和染色过程工艺条件与技术步骤;采用参与式观察法对4种色相的10个染色工艺过程进行了记录,采集了凭证标本和图像资料;对调查信息进行了整理编目。结果表明：西双版纳地区的傣族使用11种着色植物和17种助染植物;目标色相有红、黄、蓝和绿。分析了傣族染料植物资源的发掘潜力、傣族利用植物染色对于染料植物利用的应用启发。该研究详细深入地记录了西双版纳傣族使用的染料植物的种类及其相关的组合和染色的过程。该研究结果对民族民间染料植物与染色工艺的产业化应用具有重要借鉴意义,为染料植物资源筛选及其染色工艺条件优化提供了参考。     

Physiology and proteome responses of two contrasting rice mutants and their wild type parent under salt stress conditions at the vegetative stage

Salinity is one of the major environmental limiting factors that affects growth and productivity of rice (Oryza sativa L.) worldwide. Rice is among the most sensitive crops to salinity, especially at early vegetative stages. In order to get a better understanding of molecular pathways affected in rice mutants showing contrasting responses to salinity, we exploited the power of 2-DE based proteomics to explore the proteome changes associated with salt stress response. Our physiological observations showed that standard evaluation system (SES) scores, Na⁺ and K⁺ concentrations in shoots and Na⁺/K⁺ ratio were significantly different in contrasting mutants under salt stress condition. Proteomics analysis showed that, out of 854 protein spots which were reproducibly detected, 67 protein spots showed significant responses to salt stress. The tandem mass spectrometry analysis of these significantly differentially accumulated proteins resulted in identification of 34 unique proteins. These proteins are involved in various molecular processes including defense to oxidative stresses, metabolisms, photosynthesis, protein synthesis and processing, signal transduction. Several of the identified proteins were emerged as key participants in salt stress tolerance. The possible implication of salt responsive proteins in plant adaptation to salt stress is discussed.     

Use of ion-channel modulating agents to study cyanobacterial Na<Superscript>+</Superscript>-K<Superscript>+</Superscript> fluxes

Here we describe an experimental design aimed to investigate changes in total cellular levels of Na⁺ and K⁺ ions in cultures of freshwater filamentous cyanobacteria. Ion concentrations were measured in whole cells by flame photometry. Cellular Na⁺ levels increased exponentially with rising alkalinity, with K⁺ levels being maximal for optimal growth pH (∼8). At standardized pH conditions, the increase in cellular Na⁺, as induced by NaCl at 10 mM, was coupled by the two sodium channel-modulating agents lidocaine hydrochloride at 1 μM and veratridine at 100 μM. Both the channel-blockers amiloride (1 mM) and saxitoxin (1 μM), decreased cell-bound Na⁺ and K⁺ levels. Results presented demonstrate the robustness of well-defined channel blockers and channel-activators in the study of cyanobacterial Na⁺- K⁺ fluxes. Published: June 29, 2004.     

短期NaCl胁迫对不同小麦品种幼苗K+吸收和Na+、K+积累的影响

为研究盐胁迫下小麦幼苗生长及Na⁺、K⁺的吸收和积累规律,以中国春、洲元9369和长武134等3种耐盐性不同小麦品种为材料,采用非损伤微测技术检测盐胁迫2 d后的根系K⁺离子流变化,并对植株体内的Na⁺、K⁺含量进行测定。结果表明:短期(2d)盐胁迫对小麦生长有抑制作用,且对根系的抑制大于地上部,耐盐品种下降幅度小于盐敏感品种。盐胁迫下,小麦根际的 K⁺大量外流,盐敏感品种中国春K⁺流速显著高于耐盐品种长武134,最高可达15倍。小麦幼苗地上部分和根系均表现为Na⁺积累增加,K⁺积累减少,Na⁺/K⁺比随盐浓度增加而上升。中国春限Na⁺能力显著低于长武134,Na⁺/K⁺则显著高于长武134。综上所述,盐胁迫下造成小麦组织器官中Na⁺/K⁺比上升的主要原因是根系K⁺大量外流和Na⁺的过量积累,耐盐性不同的小麦品种间差异显著,并认为根系对K⁺的保有能力可能是作物耐盐性评价的一个重要指标。     

Overexpression of a poplar two-pore K+ channel enhances salinity tolerance in tobacco cells

Populus euphratica is a plant model intensively studied for elucidating physiological and molecular mechanisms of salt tolerance in woody species. Several studies have shown that vacuolar potassium (K⁺) ion channels of the two-pore K⁺ (TPK) family play an important role in maintaining K⁺ homeostasis. Here, we cloned a putative TPK channel gene from P. euphratica, termed PeTPK. Sequence analysis of PeTPK1 identified the universal K-channel-specific pore signature, TXGYGD. Over-expression of PeTPK1 in tobacco BY-2 cells improved salt tolerance, but did not enhance tolerance to hyperosmotic stress caused by mannitol (200?C600?mM). After 3?weeks of NaCl stress (100 and 150?mM), PeTPK1-transgenic cells had higher fresh and dry weights than wild-type cells. Salt treatment caused significantly higher Na⁺ accumulation and K⁺ loss in wild-type cells compared to transgenic cells. During short-term salt stress (100?mM NaCl, 24-h), PeTPK1-transgenic cells showed higher cell viability and reduced membrane permeabilization compared to wild-type cells. Scanning ion-selective electrode data revealed that salt-shock elicited a significantly higher transient K⁺ efflux from PeTPK1-transgenic callus cells and protoplasts compared to that observed in wild-type cells and protoplasts. We concluded that salt tolerance in P. euphratica is most likely mediated through PeTPK1. We propose that, under salt stress, PeTPK1 functions as an outward-rectifying, K⁺ efflux channel in the vacuole that transfers K⁺ to the cytosol to maintain K⁺ homeostasis.     

Herbivore exposure alters ion fluxes and improves salt tolerance in a desert shrub

Plants have evolved complex mechanisms that allow them to withstand multiple environmental stresses, including biotic and abiotic stresses. Here, we investigated the interaction between herbivore exposure and salt stress of Ammopiptanthus nanus, a desert shrub. We found that jasmonic acid (JA) was involved in plant responses to both herbivore attack and salt stress, leading to an increased NaCl stress tolerance for herbivore-pretreated plants and increase in K⁺/Na⁺ ratio in roots. Further evidence revealed the mechanism by which herbivore improved plant NaCl tolerance. Herbivore pretreatment reduced K⁺ efflux and increased Na⁺ efflux in plants subjected to long-term, short-term, or transient NaCl stress. Moreover, herbivore pretreatment promoted H⁺ efflux by increasing plasma membrane H⁺-adenosine triphosphate (ATP)ase activity. This H⁺ efflux creates a transmembrane proton motive force that drives the Na⁺/H⁺ antiporter to expel excess Na⁺ into the external medium. In addition, high cytosolic Ca²⁺ was observed in the roots of herbivore-treated plants exposed to NaCl, and this effect may be regulated by H⁺-ATPase. Taken together, herbivore exposure enhance s A. nanus tolerance to salt stress by activating the JA-signalling pathway, increasing plasma membrane H ⁺ - ATPase activity, promoting cytosolic Ca²⁺ accumulation, and then restricting K⁺ leakage and reducing Na⁺ accumulation in the cytosol.     

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⁺...     

Ionic and osmotic components of salt stress specifically modulate net ion fluxes from bean leaf mesophyll

Ionic mechanisms of salt stress perception were investigated by non‐invasive measurements of net H⁺, K⁺, Ca²⁺, Na⁺, and Cl^? fluxes from leaf mesophyll of broad bean (Vicia faba L.) plants using vibrating ion‐selective microelectrodes (the MIFE technique). Treatment with 90 m M NaCl led to a significant increase in the net K⁺ efflux and enhanced activity of the plasma membrane H⁺‐pump. Both these events were effectively prevented by high (10 m M ) Ca²⁺ concentrations in the bath. At the same time, no significant difference in the net Na⁺ flux has been found between low‐ and high‐calcium treatments. It is likely that plasma membrane K⁺ and H⁺ transporters, but not the VIC channels, play the key role in the amelioration of negative salt effects by Ca²⁺ in the bean mesophyll. Experiments with isotonic mannitol application showed that cell ionic responses to hyperosmotic treatment are highly stress‐specific. The most striking difference in response was shown by K⁺ fluxes, which varied from an increased net K⁺ efflux (NaCl treatment) to a net K⁺ influx (mannitol treatment). It is concluded that different ionic mechanisms are involved in the perception of the ‘ionic’ and ‘osmotic’ components of salt stress.     

Functional Characterization of a Wheat NHX Antiporter Gene TaNHX2 That Encodes a K+/H+ Exchanger

The subcellular localization of a wheat NHX antiporter, TaNHX2, was studied in Arabidopsis protoplasts, and its function was evaluated using Saccharomyces cerevisiae as a heterologous expression system. Fluorescence patterns of TaNHX2-GFP fusion protein in Arabidopsis cells indicated that TaNHX2 localized at endomembranes. TaNHX2 has significant sequence homology to NHX sodium exchangers from Arabidopsis, is abundant in roots and leaves and is induced by salt or dehydration treatments. Western blot analysis showed that TaNHX2 could be expressed in transgenic yeast cells. Expressed TaNHX2 protein suppressed the salt sensitivity of a yeast mutant strain by increasing its K⁺ content when exposed to salt stress. TaNHX2 also increased the tolerance of the strain to potassium stress. However, the expression of TaNHX2 did not affect the sodium concentration in transgenic cells. Western blot analysis for tonoplast proteins indicated that the TaNHX2 protein localized at the tonoplast of transgenic yeast cells. The tonoplast vesicles from transgenic yeast cells displayed enhanced K⁺/H⁺ exchange activity but very little Na^+/H⁺ exchange compared with controls transformed with the empty vector; Na⁺/H⁺ exchange was not detected with concentrations of less than 37.5 mM Na⁺ in the reaction medium. Our data suggest that TaNHX2 is a endomembrane-bound protein and may primarily function as a K⁺/H⁺ antiporter, which is involved in cellular pH regulation and potassium nutrition under normal conditions. Under saline conditions, the protein mediates resistance to salt stress through the intracellular compartmentalization of potassium to regulate cellular pH and K⁺ homeostasis.     

Cation co-tolerance phenomenon in cell cultures of Oryza sativa adapted to LiCl and NaCl

Cell lines of Oryza sativa L. (cv. Taipei-309) were adapted to 30 mM LiCl and 150 mM NaCl. Both adapted lines were considerably more tolerant than non adapted line when grown on 200, 250 and 300 mM NaCl and 30 mM LiCl stresses. The tolerance of LiCl-adapted line to NaCl (150 to 300 mM) and the tolerance of NaCl-adapted cells line to LiCl (30 mM) indicated that there was a cross-adaptation towards alkali metals (Na⁺ and Li⁺) not the Cl^–. Na⁺ and K⁺ contents of all lines which increased with increasing medium salinity but to a different degree. The increase in Na⁺ and K⁺ content in NaCl-adapted and non-adapted lines were comparable, while LiCl-adapted line accumulated significantly lower Na⁺and higher K⁺ content. Proline content of all lines increased with the increase in NaCl-stress but the magnitude of increase was much higher in the LiCl-adapted than other lines. The differential response of adapted lines to NaCl stress in accumulating proline and maintaining the ionic contents reveals that adapted lines have evolved different features of adaptation to cope with NaCl stress.     

Effects of NaCl and mannitol induced stress on sugarcane (<Emphasis Type="Italic">Saccharum</Emphasis> sp.) callus cultures

The effects of NaCl and mannitol iso-osmotic stresses on calli issued from sugarcane cultivars (cvs.) R570, CP59-73 and NCo310 were investigated in relation to callus growth, water content, ion and proline concentrations. Callus growth and water content decreased under both stresses with the highest reduction under mannitol-induced osmotic stress. The ion concentration was drastically affected after exposure to NaCl and mannitol. Salt stress induced an increase in Na⁺ and Cl⁻ accumulation and a decrease in K⁺ and Ca²⁺ concentrations. Under mannitol-induced osmotic stress, K⁺ and Ca²⁺ concentrations decreased significantly while Na⁺ and Cl⁻ concentrations remained unchanged. Free proline accumulation occurred under both stresses and was more marked in stress-sensitive cv. than in stress-resistant one. Our results indicated that the physiological mechanisms operating at the plant cell level in response to salt- and osmotic-induced stress in sugarcane cvs. are different. Among the cvs., we concluded that the stress resistance is closely related to the maintain of an adequate water status and a high level of K⁺ and Ca²⁺ under both stresses and a low level of Na⁺ concentration in the presence of NaCl. Thus, sugarcane (Saccharum sp.) can be regarded as a Na⁺ excluder. We also provided evidence that proline accumulation is a stress-sensitive trait rather than a stress resistance marker.     

Extracellular ATP Signaling Is Mediated by H2O2 and Cytosolic Ca2+ in the Salt Response of Populus euphratica Cells

Extracellular ATP (eATP) has been implicated in mediating plant growth and antioxidant defense; however, it is largely unknown whether eATP might mediate salinity tolerance. We used confocal microscopy, a non-invasive vibrating ion-selective microelectrode, and quantitative real time PCR analysis to evaluate the physiological significance of eATP in the salt resistance of cell cultures derived from a salt-tolerant woody species, Populus euphratica. Application of NaCl (200 mM) shock induced a transient elevation in [eATP]. We investigated the effects of eATP by blocking P2 receptors with suramin and PPADS and applying an ATP trap system of hexokinase-glucose. We found that eATP regulated a wide range of cellular processes required for salt adaptation, including vacuolar Na⁺ compartmentation, Na⁺/H⁺ exchange across the plasma membrane (PM), K⁺ homeostasis, reactive oxygen species regulation, and salt-responsive expression of genes related to K⁺/Na⁺ homeostasis and PM repair. Furthermore, we found that the eATP signaling was mediated by H₂O₂ and cytosolic Ca²⁺ released in response to high salt in P. euphratica cells. We concluded that salt-induced eATP was sensed by purinoceptors in the PM, and this led to the induction of downstream signals, like H₂O₂ and cytosolic Ca²⁺, which are required for the up-regulation of genes linked to K⁺/Na⁺ homeostasis and PM repair. Consequently, the viability of P. euphratica cells was maintained during a prolonged period of salt stress.     

Photosynthesis,chlorophyll fluorescence,inorganic ion and organic acid accumulations of sunflower in responses to salt and salt-alkaline mixed stress

Sunflowers were treated with mixing proportions of NaCl, Na₂SO₄, NaHCO₃, and Na₂CO₃. Effects of salt and saltalkaline mixed stress on growth, photosynthesis, chlorophyll fluorescence, and contents of inorganic ions and organic acids of sunflower were compared. The growth of sunflower decreased with increasing salinity. The contents of photosynthetic pigments did not decrease under salt stress, but their contents decreased sharply under salt-alkaline mixed stress. Net photosynthetic rates, stomatal conductance and intercellular CO₂ concentration decreased obviously, with greater reductions under salt-alkaline mixed stress than under salt one. Fluorescence parameters showed no significant differences under salt stress. However, maximal efficiency of PSII photochemistry, photochemical quenching coefficient, electron transport rate, and actual PSII efficiency significantly decreased but non-photochemical quenching increased substantially under salt-alkaline mixed stress. Under salt-alkaline mixed stress, sunflower leaves maintained a low Na⁺- and high K⁺ status; this may be an important feature of sunflower tolerance to salinity. Analysis of the mechanism of ion balance showed that K⁺ but not Na⁺ was the main inorganic cation in sunflower leaves. Our results indicated that the change in organic acid content was opposite to the change of Cl⁻, and the contribution of organic acid to total charge in sunflower leaves under both stresses decreased with increasing salinity. This may be a special adaptive response to stresses for sunflower. Sunflower under stress conditions mainly accumulated inorganic ions instead of synthesizing organic compounds to decrease cell water potential in order to save energy consumption.