小麦耐盐细胞系对盐胁迫的伤害性反应

通过逐级提高ＮａＣｌ浓度的筛选方法,得到了能在１．５％ＮａＣｌ下生长良好的小麦（ＴｒｉｔｉｃｕｍａｅｓｔｉｖｕｍＬ．）耐盐细胞系。在盐分胁迫下,耐盐细胞系含水量的降低幅度小于不耐盐细胞系（对照）,Ｈ２Ｏ２含量和Ｏ－２产生速率的增加幅度也明显小于对照细胞系。同时,膜的相对透性、膜脂过氧化和脱酯化程度的提高幅度也明显低于对照细胞系。表明盐分对小麦细胞系膜的伤害与活性氧介导的膜脂过氧化和脱酯化有关,而耐盐细胞系比对照细胞系表现出较强的抗活性氧伤害的能力。     

Changes in plasma membrane lipids, aquaporins and proton pump of broccoli roots, as an adaptation mechanism to salinity

Salinity stress is known to modify the plasma membrane lipid and protein composition of plant cells. In this work, we determined the effects of salt stress on the lipid composition of broccoli root plasma membrane vesicles and investigated how these changes could affect water transport via aquaporins. Brassica oleracea L. var. Italica plants treated with different levels of NaCl (0, 40 or 80 mM) showed significant differences in sterol and fatty acid levels. Salinity increased linoleic (18:2) and linolenic (18:3) acids and stigmasterol, but decreased palmitoleic (16:1) and oleic (18:1) acids and sitosterol. Also, the unsaturation index increased with salinity. Salinity increased the expression of aquaporins of the PIP1 and PIP2 subfamilies and the activity of the plasma membrane H⁺-ATPase. However, there was no effect of NaCl on water permeability (P_f) values of root plasma membrane vesicles, as determined by stopped-flow light scattering. The counteracting changes in lipid composition and aquaporin expression observed in NaCl-treated plants could allow to maintain the membrane permeability to water and a higher H⁺-ATPase activity, thereby helping to reduce partially the Na⁺ concentration in the cytoplasm of the cell while maintaining water uptake via cell-to-cell pathways. We propose that the modification of lipid composition could affect membrane stability and the abundance or activity of plasma membrane proteins such as aquaporins or H⁺-ATPase. This would provide a mechanism for controlling water permeability and for acclimation to salinity stress.     

Plasma membrane lipid composition of the halophilic cyanobacterium Aphanothece halophytica

The plasma membrane from Aphanothece halophytica was isolated using both glycerol and sucrose gradient centrifugation. The isolated membrane was characterized for lipid content by TLC and isolated lipids were quantified by chemical analysis. The plasma membrane of A. halophytica was composed of MGDG, DGDG and PG. The sulfur containing lipid SQDG was not detected. The mole percent of each lipid in the plasma membrane varied with the external salinity of the media. MGDG was the most abundant lipid in the plasma membrane of cells grown at one molar external NaCl. At three molar external NaCl, PG was the most abundant lipid. The ratio of uncharged to charged lipids comprising the plasma membrane decreased as the external salinity increased. It is possible that the alteration in lipid composition is of major importance in the adaptation of A. halophytica to changing external salinity.Abbreviations TLC Thin-layer chromatography - MGDG momogalactosyldiacylglycerol - DGDG digaloctosyldiacylglycerol - PG phosphatidylglycerol - SQDG sulphoquinovosyldiacylglycerol     

Osmotic water permeability and solute reflection coefficients of rat kidney brush-border membrane vesicles

Solute reflection coefficients, sigma i, of rat kidney brush-border membrane vesicles were determined by the comparison of water flows induced by equiosmolal gradients of sucrose and NaCl, KCl or mannitol. The values of 0.53 for sigma NaCl and 0.56 for sigma KCl when compared with 0.92 for sigma mannitol suggested some interactions between salt and water pathways. Altering the membrane proteins with 0.4 mM HgCl2 decreased the osmotic water permeability of the vesicles by 70 to 80% and brought sigma NaCl and sigma KCl to a value not different from 1. This argued in favor of water protein pathways in the luminal membrane of kidney proximal cells which are partly accessible to NaCl and KCl.     

Variation in polar-group content in lipids of cowpea (Vigna unguiculata) Cell cultures as a mechanism of haloadaptation

Callus and cell suspension cultures of cowpea (Vigna unguiculata) were induced with 2,4-dichlorophenoxyacetic acid and grown at different NaCl concentrations. The cell biomass yield and its total lipid content decreased with increasing salinity. However, while the hexose content in lipids was higher, the amount of lipid phosphorus was significantly lower in both agar and cell suspension cultures. Ion-transport rates with artificial membranes prepared with different lipid fractions showed that lipids from cells grown in a saline medium were less permeable to Na⁺ and to Cl^- than those grown in a non-saline medium. Also the permeability of membranes prepared with glycolipids was lower than those prepared with phospholipids and whole lipids. Apparently, the increase of hexose/phosphorus ratio in membrane lipids is induced in response to the halo-adaptation process.     

NaCl胁迫对小黄花菜生长及相关生理指标的影响

为探讨小黄花菜的耐盐机理,选育良好的耐盐植物以缓解土壤盐渍化问题,该文选取小黄花菜(Hemerocallis minor)为试材,采用砂培法,研究不同浓度Na Cl(50、100、150、200、250 mmol·L~(-1))胁迫对小黄花菜的生长性状、细胞质膜透性和有机渗透调节物质含量等的影响。结果表明:(1)小黄花菜在100～150mmol·L~(-1)Na Cl胁迫时,损害初步显现,但不影响其存活;在Nacl浓度为200 mmol·L~(-1)以上时,小黄花菜生长被显著抑制,造成根系不发育、叶片受害、植株干物质积累显著不足,严重影响其生存状态。(2)在50～150mmol·L~(-1)盐渍环境下,叶片膜透性、MDA含量增幅较小,该浓度范围的Na Cl胁迫造成的膜脂损伤有限; 200mmol·L~(-1)以上浓度的Na Cl胁迫使得小黄花菜叶片的离子平衡无法继续维持,膜的选择透性丧失。(3)随着Na Cl浓度的增加,叶片中脯氨酸含量显著增加;在50～100 mmol·L~(-1)Nacl胁迫下,叶片可溶性糖含量在胁迫初期有所增加,在15 d时达到最大,胁迫后期开始下降;叶片中可溶性蛋白含量的变幅较为平缓,说明小黄花菜的主要渗透调节物质不是可溶性蛋白。该研究发现通过提高叶片膜透性,促进自身有机渗透调节物质的合成,能够在一定程度上缓解盐渍对植株的侵害,使得小黄花菜能够在50～100 mmol·L~(-1)的盐碱环境下正常生长。     

Aluminum and Temperature Alteration of Cell Membrane Permeability of Quercus rubra

This report extends research on Al-induced changes in membrane behavior of intact root cortex cells of Northern red oak (Quercus rubra). Membrane permeability was determined by the plasmometric method for individual intact cells at temperatures from 2 or 4 to 35°C. Al (0.37 millimolar) significantly increased membrane permeability to urea and monoethyl urea and decreased permeability to water. Al significantly altered the activation energy required to transport water (+32%), urea (+9%), and monoethyl urea (−7%) across cell membranes. Above 9°C, Al increased the lipid partiality of the cell membranes; below 7°C, Al decreased it. Al narrowed by 6°C the temperature range over which plasmolysis occurred without membrane damage. These changes in membrane behavior are explainable if Al reduces membrane lipid fluidity and kink frequency and increases packing density and the occurrence of straight lipid chains.     

Characterization of cell surface material removed by water and NaCl washing ofParacoccus denitrificans grown under conditions of divalent cation deficiency and sufficiency

Paracoccus denitrificans grown on complex medium deficient in Mg²⁺ and Ca²⁺ are rendered lysozyme susceptible by washing with NaCl, whereas cells grown in a succinate-salts medium (Mg²⁺ and Ca²⁺ sufficient) or complex medium supplemented with Mg²⁺+Ca²⁺ are not. The material released by water washing of cells grown on complex medium and complex medium supplemented with Mg²⁺ and Ca²⁺ was characterized by a high protein content. There was a high lipid: protein ratio and an appreciable amount of 3-deoxyoctulosonic acid in the material released by NaCl washing of cells grown under all conditions, indicating release of outer membrane material. The lipid ornithine: lipid phosphorous ratios of NaCl wash from cells grown on complex medium and complex medium supplemented with Mg²⁺ and Ca²⁺ were 0.54 and 0.34, respectively. Although NaCl washing removed outer membrane material from cells grown under all conditions, only divalent cation deficient cells were rendered lysozyme susceptible. This might be explained by the increased outer membrane ornithine-containing lipid to phospholipid ratio in these cells yielding a more permeable outer membrane.     

Phallolysin A mushroom toxin,forms proton and voltage gated membrane channels

Phallolysin, a water soluble protein of M _r 34,000 produced by the poisonous mushroom Amanita phalloides, causes lysis of various mammalian cell types. Lysis is thought to be initiated by the formation of ion permeable membrane channels. We therefore studied the interaction of phallolysin with solvent-free planar lipid bilayers. In the presence of low phallolysin concentrations (10–100 nM) single channel current fluctuations were observed. Unit channel conductances are 44 pS in 500 mM NaCl and 77 pS in 1 M NaCl. Although the channel does not significantly discriminate between alkali cations, its permeability to Cl^- is lower (P _K ⁺/P _Cl ^-=4/1). Gating kinetics display a pronounced bursting behavior and a dependence on membrane voltage, cis side pH-value, and on membrane lipid composition. An equivalence relation between membrane voltage and proton concentration was found, i.e. a pH change of one unit is equivalent to a corresponding voltage change of 130 mV. Dependence on the amount of negatively charged lipids is explained by changes of the actual pH due to surface charge effects.Abbreviations 1,3-SMPC 1-stearoyl-3-myristoyl-glycero-2-phosphocholine - 1,2-DOPS 1,2-dioleoyl-glycero-3-phosphoserine     

盐诱导的依赖叶黄素循环的热耗散提高了小麦的耐盐性

以小麦抗盐品种‘DK961’和盐敏感品种‘LM15’为材料,探讨盐胁迫条件下叶黄素循环与膜脂过氧化的关系。结果表明：200mmol·L-1NaCl处理后二者地上部分鲜重、含水量、K＋含量显著下降,Na＋含量、Na＋/K＋比、丙二醛含量显著升高,膜透性显著增大,‘LM15’的变化幅度均明显大于‘DK961’,而‘LM15’脱环氧化状态（A＋Z）/（A＋Z＋V）的增加明显小于‘DK961’。这表明盐胁迫下‘DK961’通过增加依赖叶黄素循环的热耗散减轻了膜脂过氧化,提高其耐盐性。     

等渗Ca(NO3)2和NaCl胁迫对黄瓜幼苗生长及渗透调节物质含量的影响

采用营养液培养方法,以耐盐性较弱的‘津春2号’黄瓜品种为试材,研究了等渗Ca(NO3)2和NaCl胁迫对黄瓜幼苗生长、根系电解质渗透率、根系活力、Na+和K+含量及渗透调节物质含量的影响。结果显示:(1)在84mmol.L-1 NaCl和56mmol.L-1 Ca(NO3)2等渗胁迫下,黄瓜幼苗鲜重和干重均显著下降,且NaCl处理下降的幅度大于等渗Ca(NO3)2处理。(2)NaCl主要通过对黄瓜根系的伤害来抑制植株生长,表现为根系活力下降、根系质膜透性增大、Na+大量积累、K+含量显著下降、Na+/K+明显上升,最终导致根冠比下降;而Ca(NO3)2处理对根系质膜透性、K+含量、Na+/K+的影响均小于NaCl胁迫,且根系活力和根冠比上升,但Ca(NO3)2胁迫后叶片含水量和渗透调节能力均小于NaCl胁迫。(3)NaCl胁迫条件下,黄瓜幼苗内渗透调节物质以可溶性糖为主,而Ca(NO3)2胁迫以可溶性蛋白为主。研究表明,NaCl胁迫对黄瓜幼苗的伤害大于等渗Ca(NO3)2,NaCl主要通过破坏根系质膜结构影响植株生长,而Ca(NO3)2主要通过引起地上部生理干旱来影响植株生长。     

Effect of retinol and retinoic acid on permeability,electrical resistance and phase transition of lipid bilayers

Retinol and retinoic acid have been incorporated into the artificial membrane systems, planar bimolecular lipid membranes and liposomes, and their effects on several membrane parameters have been measured. 1. Retinol and retinoic acid increased the permeability of egg lecithin liposomes to K⁺, I^? and glucose when incorporated into the membranes at levels as low as 0.5 membrane mol%. Retinoic acid influenced permeability more than did retinol for each of the solutes tested. 2. Retinol and retinoic acid both decreased the electrical resistance of egg lecithin-planar bimolecular lipid membranes from 0.5 to 8 membrane mol%. Retinoic acid effected a larger change than did retinol. 3. Retinol and retinoic acid increased the permeability of dimyristoylphosphatidylcholine and dipalmitoylphosphatidylcholine liposomes to water at 1.0 and 3.0 membrane mol%. A larger effect on water permeability was measured for retinoic acid than for retinol. 4. Retinol and retinoic acid at 1.0 and 3.0 membrane mol% were shown to lower the phase-transition temperature of liposomes composed of dimyristoylphosphatidylcholine or dipalmitoylphosphatidylcholine. Phase-transition temperatures were monitored by abrupt changes in water permeability and liposome size associated with the transition. Retinoic acid lowered the phase-transition temperature of dimyristoylphosphatidylcholine liposomes more than did retinol, while both retinoids had almost the same effect on dipalmitoylphosphatidylcholine liposomes.     

Permeability behaviour of lipid vesicles prepared from plant plasma membranes--impact of compositional changes

Exposure of oat seedlings to repeated moderate water deficit stress causes a drought acclimation of the seedlings. This acclimation is associated with changes in the lipid composition of the plasma membrane of root cells. Here, plasma membranes from root cells of acclimated and control plants were isolated using the two-phase partitioning method. Membrane vesicles were prepared of total lipids extracted from the plasma membranes. In a series of tests the vesicle permeability for glucose and for protons were analysed and compared with the permeability of model vesicles. Further, the importance of critical components for the permeability properties was analysed by modifying the lipid composition of the vesicles from acclimated and from control plants. The purpose was to add specific lipids to vesicles from acclimated plants to mimic the composition of the vesicles from control plants and vice versa. The plasma membrane lipid vesicles from acclimated plants had a significantly increased permeability for glucose and decreased permeability for protons as compared to control vesicles. The results point to the importance of the ratio phosphatidylcholine (PC)/phosphatidylethanolamine (PE), the levels of cerebrosides and free sterols and the possible interaction of these components for the plasma membrane as a permeability barrier.     

Alterations in root lipid peroxidation and antioxidative responses in two rice cultivars under NaCl-salinity stress

The comparative alterations of short term NaCl stress and recovery on growth, water relations, ionic composition, lipid peroxidation and antioxidants in roots of two rice cultivars differing in salt tolerance were studied. Exposed for 24 h to increasing (50, 100 and 150 mmol l⁻¹) concentrations of NaCl, roots of 12D Oryza sativa L. cv. Lunishree and cv. Begunbitchi decreased in fresh weight, dry weight and relative water content. Increased Na⁺ and decreased K⁺ ion were determined at increasing NaCl concentrations. Both peroxide content and lipid peroxidation measured in terms of MDA level increased and the ratio was higher in Begunbitchi compared to Lunishree. Recovered roots showed lower peroxide and MDA content. Ascorbate and glutathione contents increased in the stressed and recovered roots of Lunishree, but decreased in Begunbitchi with increasing NaCl concentrations. Although SOD, CAT and GR activities decreased in the stressed roots, CAT activity also increased in recovered roots of both the cultivars. The POX activity increased in stressed and recovered roots of both Lunishree and Begunbitchi. Higher free radicals scavenging capacity and more efficient protection mechanism of Lunishree against salt stress, as revealed by the lower level of lipid peroxidation and improved plant water status as well as activities of some of the antioxidants, suggest that significant cultivar differences in response to salt stress in rice are closely related to differences in the activities of antioxidants and ion content. Another possible conclusion is that improved tolerance to salt stress may be accomplished by increased capacity of antioxidative system.     

Stopped-flow study of anesthetic effect on water-transport kinetics through phospholipid membranes. Interfacial versus lipid core ligands

We have compared ligand effects between polar and apolar anesthetic molecules upon water transport across phospholipid membranes by kinetic analysis of the osmotic swelling rate, using a stopped-flow technique. Chloroform and 1-hexanol were used as interfacial ligands, and carbon tetrachloride and n-hexane were used as their counterparts, representing lipid core action. Because anesthetics transform the solid-gel membrane into a liquid-crystalline state, and because phospholipid membranes display an anomaly in permeability at the phase transition, dimyristoylphosphatidylcholine vesicles were studied at temperatures above the main phase transition to avoid this anomaly. All these molecules increased the osmotic swelling rate. However, a significant difference was observed in the activation energy, delta Ep, between polar and apolar molecules; delta Ep was almost unaltered by the addition of polar molecules (chloroform and 1-hexanol), whereas it was decreased by apolar molecules (carbon tetrachloride and n-hexane). The obtained results were analyzed in terms of the dissolution-diffusion mechanism for water permeation across the lipid membrane. It is suggested that polar molecules affect water permeability by altering the partition of water between the membrane interior and water phase, and apolar molecules affect it by altering both the partition and the diffusion of water within the membrane interior.     

Al and Ca Alteration of Membrane Permeability of Quercus rubra Root Cortex Cells

This study was undertaken to quantify the effect of aluminum and calcium on membrane permeability. The influence of Ca²⁺ (0.2-3.7 millimolar) and Al³⁺ (0-3.7 millimolar) on the permeability of root cortical cells of Quercus rubra was measured using three nonelectrolytes (urea, methyl urea, and ethyl urea) as permeators of progressively increasing lipid solubility. Water permeability was also measured. Al³⁺ (a) increased membrane permeability to the nonelectrolytes, (b) decreased the membrane's partiality for lipid permeators, and (c) decreased membrane permeability to water. Ca²⁺ had effects on permeability that were opposite to those of Al³⁺. When Al³⁺ and Ca²⁺ were tested in combination, these opposite effects counteracted each other. The results suggest that Al³⁺ altered the architecture of membrane lipids.     

Salt tolerance inNitellopsis obtusa

Summary The mechanism of salt tolerance was studied using isolated internodal cells of the charophyteNitellopsis obtusa grown in fresh water. When 100 mM NaCl was added to artificial pond water (0.1 mM each of NaCl, KC1, CaCl₂), no cell survived for more than one day. Within the first 30 minutes, membrane potential (E_m) depolarized and membrane resistance (R_m) decreased markedly. Simultaneously, cytoplasmic Na⁺ increased and K⁺ decreased greatly. At steady state the increase in Na⁺ content was roughly equal to the decrease in K⁺ content. The Cl^– content of the cytoplasm did not change. These results suggest that Na⁺ enters the cytoplasm by exchange with cytoplasmic K⁺. Both the entry of Na⁺ and the exit of K⁺ are assumed to be passive and the latter being caused by membrane depolarization. Vacuolar K⁺, Na⁺, and Cl^– remained virtually constant, suggesting that rapid influx of Na⁺ from the cytoplasm did not occur.In 100 mM NaCl containing 10 mM CaCl₂, membrane depolarization, membrane resistance decrease and changes in cytoplasmic [Na⁺] and [K⁺] did not occur, and cells survived for many days. When cells treated with 100 mM NaCl were transferred within 1 hour to 100 mM NaCl containing 10 mM CaCl₂, Em decreased, Rm increased, cytoplasmic Na⁺ and K⁺ returned to their initial levels, and cells survived. Two possible mechanisms for the role of Ca²⁺ in salt tolerance inNitellopsis are discussed; one a reduction in plasmalemma permeability to Na⁺ and the other a stimulation of active Na⁺-extrusion.     

Influence of the growth at high osmolality on the lipid composition, water permeability and osmotic response of Lactobacillus bulgaricus

Changes in water permeability and membrane packing were measured in cells of Lactobacillus bulgaricus and in vesicles prepared with lipids extracted from them. The osmotic response of whole cells and vesicles is compared with the one of bacteria grown in a high osmolal medium. Both bacteria and vesicles, behave as osmometers. This means that the volume decrease is promoted by the outflow of water, driven by the NaCl concentration difference, arguing that neither Na+ nor Cl- permeates the cell or the lipid membrane in these conditions. Therefore, the volume changes can be correlated with the rate of water permeation across the cell or the vesicle membranes. The permeation of water was analyzed as a function of the lipid species by measuring the volume changes and the saturation ratio of the lipids. To put into relevance the membrane processes, the permeation properties of lipid vesicles prepared with lipids extracted from bacteria grown in normal and high osmolality conditions were also analyzed. The permeation response was correlated with the physical properties of the membrane of whole cells and vesicles, by means of fluorescence anisotropy of diphenyl hexatriene (DPH). The modifications in membrane properties are related with the changes in the membrane composition triggered by the growth in a high osmolal medium. The changes appear related to an increase in the sugar content of the whole pool of lipids and in the saturated fatty acid residues.     

Monovalent ion permeability of model bilayer membranes based on lipids from tissues of various vertebrates

The ionic permeability of model bilayer membranes prepared from total lipids of the brain, heart, liver, and muscles of four species of higher vertebrates was studies. The electrical conductivity and potential of membranes breakdown were measured in KCl, NaCl, and LiCl solutions. It was found that the permeability for potassium ions of bilayer lipid membranes from the lipids of nervous tissue decreases in the row of poikilothermal vertebrates and increases in mammals. A reverse regularity was observed for membrane stability, which manifested itself as an increase in membrane breakdown in the order fish-amphibian-reptile and a decrease in membrane breakdown in mammalian membranes. It was shown that, in most cases, the the permeability of bilayer lipid membranes for K+ ions is higher than for Na+ and Li+.     

Salinity tolerance of Kosteletzkya virginica. II. Root growth, lipid content, ion and water relations

Abstract. Kosteletzkya virginica (L.) Presl., a dicot halophyte native to brackish tidal marshes, was grown on nutrient solution containing 0. 85, 170 or 255 mol m ³ NaCl, and the effects of external salinity on root growth, ion and water levels, and lipid content were examined in successive harvests. Root growth paralleled shoot growth trends, with some enhancement observed at 85 mol m ³ NaCl and a reduction noted at the higher salinities. Root Na⁺ content increased with increasing external NaCl, but remained constant with time for each treatment. K⁺ content, although lower in salt-grown plants after 14 d salinization, subsequently increased to levels comparable to unsalinized plants. A strong K⁺ affinity was reflected in the increased K⁺/Na⁺ selectivity of salt-grown plants and by their low Na⁺/K⁺ ratios. Cl levels rose in salinized plants and values were double or more those for Na⁺, indicating the possibility of a sodium-excluding mechanism in roots. Root phospholipids and sterols, principal membrane constituents, were maintained or elevated and the free sterol/phospholipids ratio increased in salinized K. virginica plants, suggesting retention of overall membrane structure and decreased permeability. This response, considered in light of root calcium maintenance and high potassium levels, suggests that salinity-induced changes in membrane lipid composition may be important in preventing K⁺ leakage from cells.