Fatty acid composition of lipids in vegetative organs of the halophyte Suaeda altissima under different levels of salinity

Qualitative and quantitative composition of fatty acids (FA) in the lipids of vegetative organs of the halophyte Suaeda altissima (L.) Pall. grown at different NaCl concentrations in nutrient solution was studied. Along with this, the biomass of these organs, the content of water and Na⁺, Cl^?, and K⁺ ions in them, and the ultrastructure of root and leaf cells were determined. At both low (1 mM) and high (750 mM) NaCl concentrations in nutrient solution, plants could maintain growth and water content in organs, demonstrating a noticeable increase in the dry weight and a slight increase in the water content at 250 mM NaCl. At all NaCl concentrations in nutrient solution, S. altissima tissues contained a relatively high K⁺ amount. Under salinity, Na⁺ and Cl^? ions contributed substantially into the increase in the cell osmotic pressure, i.e., a decrease in their water potential; in the absence of salinity, K⁺ fulfilled this function. In the cells of both roots and leaves, NaCl stimulated endo- and exocytosis, supposedly involved in the vesicular compound transport. 750 mM NaCl induced plasmolysis and changes in the membrane structure, which can be interpreted as degradation processes. Under optimal NaCl concentration in medium (250 mM), the content of lipids in plant aboveground organs per fresh weight was more than 2.5-fold higher than under 1 or 750 mM NaCl, whereas in the roots opposite patten was observed. When plants were grown under non-optimal conditions, substantial changes occurred in the qualitative and quantitative FA composition in lipids of both aboveground organs and roots. Observed changes are discussed in relation to processes underlying S. altissima salt tolerance and those of disintegration occurring at the high external NaCl concentration (750 mM).     

Conversion of quaternary ammonium compounds in the whole plant system of the halophyte Suaeda altissima

The total content of quaternary ammonium compounds (QAC) and the choline content in roots, leaves, and xylem exudates of the halophyte Suaeda altissima (L.) Pall. were determined after growing plants at various NaCl concentrations in the nutrient solution (1, 50, 100, and 250 mM). Based on the results obtained, the content of glycine betaine in organs and xylem exudates of S. altissima was estimated as the difference between the total content of QAC and the choline content. In roots choline accounted for the largest portion of QAC (from 69 to 96% at various NaCl concentrations in nutrient media), whereas in leaves it contributed only 12–23%. The contribution of choline to QAC content in the xylem exudates was 84–90%. It is concluded that choline in S. altissima is mainly synthesized in roots and is delivered with the ascending water flow to leaves where it is utilized as a substrate for glycine betaine synthesis. The content of glycine betaine in leaves increased with elevation of NaCl concentration in the nutrient solution, thus contributing appreciably to the maintenance of osmotic balance in the cytoplasm of S. altissima at high salinity.     

In vitro salt tolerance of cell wall enzymes from halophytes and glycophytes

The halophyte Suaeda maritima grows optimally in high concentrations(40–60% seawater) of salt. In these conditions the concentrationof salt in the apoplast of the leaves is at least 500 mM, aconcentration which severely inhibits the activity of cytoplasmicenzymes of both glycophytes and halophytes. The in vitro salttolerance of a number of cell wall enzymes was assayed in thepresence of a range of concentrations of NaCl. There was nosignificant inhibition of the activity of galactosidase, glucosidase,peroxidase or xyloglucan endo-transglycosylase extracted fromSuaeda maritima by in vitro concentrations of NaCl up to atleast 1 M. In vitro salt tolerance of cell wall enzymes wasnot restricted to the halophyte, similar enzymes from the non-halophilicrelative Kochia tricophylla, and from the glycophytes Vignaradiata and Cicer arietinum, were inhibited little, or not atall, by the same concentrations of salt. Pectin esterase wassomewhat less tolerant, but activity at 500 mM NaCl was stillgreater than at 0 mM NaCl in both Suaeda and Vigna. It is concludedthat these enzymes of the cell wall compartment are much moresalt-tolerant than cytoplasmic enzymes of higher plants. Theresults are discussed in relation to conditions thought to pertainin the apoplast. Key words: Apoplast, cell wall enzymes, halophyte, salt tolerance, Suaeda maritima     

Pinocytosis in the root cells of a salt-accumulating halophyte <Emphasis Type="Italic">Suaeda altissima</Emphasis> and its possible involvement in chloride transport

Ultrastructure of root cells in salt-accumulating halophyte Suaeda altissima (L.) Pall. was examined with transmission electron microscopy. Plants were grown hydroponically on nutrient media containing 3, 50, 250, and 500 mM NaCl. Some plants were exposed to hypersomotic salt shock by an abrupt increase in NaCl concentration from 50 to 400 mM. Growing S. altissima plants at high NaCl concentrations induced the formation of type 1 pinocytotic structures in root cells. Type 1 structures appeared as pinocytotic invaginations of two membranes, the plasmalemma and tonoplast. These invaginations into vacuoles gave rise to freely ‘floating’ multivesicular bodies (MVB) enclosed by a double membrane layer. The pinocytotic invaginations and MVB contained the plasmalemma-derived vesicles and membranes of endosome origin. The hyperosmotic salt shock led to formation of type 2 and type 3 pinocytotic structures. The type 2 structures were formed as pinocytotic invaginations of the tonoplast and gave rise to MVB in vacuoles. Unlike type 1 MVB, the type 2 MVB had only one enclosing membrane, the tonoplast. The type 3 structures appeared as the plasmalemma-derived vesicles located in the periplasmic space. The cytochemical electron-microscopy method was applied to determine the intracellular Cl^? localization. This method, based on sedimentation of electron-dense AgCl granules in tissues treated with silver nitrate, showed that the pinocytotic structures of all types contain Cl^? ions. The presence of Cl^? in pinocytotic structures implies the involvement of these structures in Cl^? transport between the apoplast, cytoplasm, and the vacuole.     

Sea fennel (<Emphasis Type="Italic">Crithmum maritimum</Emphasis> L.) under salinity conditions: a comparison of leaf and root antioxidant responses

The present study was carried out to compare the effect of NaCl on growth, cell membrane damage, and antioxidant defences in the halophyte Crithmum maritimum L. (sea fennel). Physiological and biochemical changes were investigated under control (0 mM NaCl) and saline conditions (100 and 300 mM NaCl). Biomass and growth of roots were more sensitive to NaCl than leaves. Roots were distinguished from leaves by increased electrolyte leakage and high malondialdehyde (MDA) concentration. Superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX) activities, ascorbic acid (AA) and glutathione (GSH) concentrations were lower in the roots than in the leaves of control plants. The different activity patterns of antioxidant enzymes in response to 100 and 300 mM NaCl indicated that leaves and roots reacted differently to salt stress. Leaf CAT, APX and glutathione reductase (GR) activities were lowest at 300 mM NaCl, but they were unaffected by 100 mM NaCl. Only SOD activity was reduced in the latter treatment. Root SOD activity was significantly decreased in response to 300 mM NaCl and root APX activity was significantly higher in plants treated with 100 and 300 mM compared to the controls. The other activities in roots were insensitive to salt. The concentration of AA decreased in leaves at 100 and 300 mM NaCl, and in roots at 300 mM NaCl, when compared to control plants. The concentrations of GSH in NaCl-treated leaves and roots were not significantly different from the controls. In both organs, AA and GSH were predominating in the total pool in ascorbic acid and glutathione, under control or saline conditions.     

Involvement of Long-Distance Na<Superscript>+</Superscript> Transport in Maintaining Water Potential Gradient in the Medium-Root-Leaf System of a Halophyte <Emphasis Type="Italic">Suaeda altissima</Emphasis>

The aim of this study was to determine the range of NaCl concentrations in the nutrient solution that allow Suaeda altissima (L.) Pall., a salt-accumulating halophyte, to maintain the upward gradient of water potential in the “medium-root-leaf” system. We evaluated the contribution of Na⁺ ions in the formation of water potential gradient and demonstrated that Na⁺ loading into the xylem is involved in this process. Plants were grown in water culture at NaCl concentrations ranging from zero to 1 M. The water potential of leaf and root cells was measured with the method of isopiestic thermocouple psychrometry. When NaCl concentration in the growth medium was raised in the range of 0–500 mM (the medium water potential was lowered accordingly), the root and leaf cells of S. altissima decreased their water potential, thus promoting the maintenance of the upward water potential gradient in the medium-root-leaf system. Growing S. altissima at NaCl concentrations f 750 mM and 1 M disordered water homeostasis and abolished the upward gradient of water potential between roots and leaves. At NaCl concentrations of 0–250 mM, the detached roots of S. altissima were capable of producing the xylem exudate. The concentration of Na⁺ in the exudate was 1.3 to 1.6 times higher than in the nutrient medium; the exudate pH was acidic and was lowered from 5.5 to 4.5 with the rise in the salt concentration. The results indicate that the long-distance Na⁺ transport and, especially, the mechanism of Na⁺ loading into the xylem play a substantial role in the formation of water potential gradient in S. altissima. The accumulation of Na⁺ in the xylem and acidic pH values of the xylem sap suggest that Na⁺ loading into the xylem is carried out by the Na⁺/H⁺ antiporter of the plasma membrane in parenchymal cells of the root stele.__________Translated from Fiziologiya Rastenii, Vol. 52, No. 4, 2005, pp. 549–557.Original Russian Text Copyright © 2005 by Balnokin, Kotov, Myasoedov, Khailova, Kurkova, Lun’kov, Kotova.     

Effect of nitrogen deficiency on the ion-exchange properties of cell wall polymers from wheat roots

The ion-exchange properties of cell wall polymers isolated from the roots of wheat (Triticum aestivum L.) plants grown on either nitrate-free (N-deficient) or nitrate-containing (+N) hydroponic nutrient medium have been investigated. Irrespective of the nitrogen nutrition regimen, the studied cell walls contained four types of ion-exchange groups: primary amino groups of structural proteins (pK_a < 3), carboxyl groups of polygalacturonic acid in pectin (pK _a ~4.7), carboxyl groups of hydroxycinnamic acids (pK _a ~7.3), and phenolic OH-groups of lignin (pK_a ~10.2). The quantitative ratio between these types of ion-exchange groups, the mass fraction of cell walls in the dry weight of roots, and the swelling coefficient of cell walls depended on the nitrate presence in the growing medium. Compared to the +N variant, the N-deficient variant was characterized by a 2.4 times higher content of phenolic OH-groups in cell walls and 1.24 times higher mass fraction of cell walls; at the same time, the swelling coefficient for this variant was lower by 10%. The obtained data indicate that nitrogen deficiency results in a formation of thicker root cell walls with a higher degree of polymer cross-linking that may be caused by the increased lignin content.     

3种荒漠盐生植物根系及根毛形态特征的比较研究

在水培条件下,针对梭梭、囊果碱蓬和钠猪毛菜3种荒漠盐生植物,研究它们苗期在不同盐浓度条件下根系和根毛形态的差异。结果表明：一定浓度的盐分可以促进3种盐生植物生长,但较高浓度的盐抑制其生长,特别是对根系生长的抑制作用更大。在同样盐浓度下,钠猪毛菜的生长最快,生物量也最大。在盐分浓度较低时,3种盐生植物的主根长和总根长都有所增加,与对照相比,囊果碱蓬增加的幅度较大,但高浓度的盐会抑制根系总长度的增加,其中囊果碱蓬较梭梭和钠猪毛菜抑制的程度轻。盐分对3种植物的根系平均直径没有显著的影响,但有减小的趋势。在水培条件下,梭梭和囊果碱蓬的根系上、中、下部分布的较均匀,而钠猪毛菜的根系中部比上部和下部有显著的增加,盐分对每种植物的根系的分布没有显著的影响。3种植物的根毛与根系生长环境中的盐浓度没有明显相关性;3种植物之间,根毛的长度和密度有显著差异。从根系和根毛的形态特征可以推断：囊果碱蓬比梭梭和钠猪毛菜具有较强的抗旱性和耐极薄能力;梭梭的耐盐能力较其它2种植物差,囊果碱蓬的耐盐性最强。     

La glutamine synthètase du Suaeda maritima. Action in vivo et in vitro du NaCl

The glutamine synthetase of Suaeda maritima. In vivo and in vitro action of NaCl Glutamine synthetase (GS; EC 6.3.1.2) was isolated and characterized from roots and aerial parts of the halophyte Suaeda maritima (L.) Dum. var. macrocarpa Moq. Km values of GS were identical in both types of organ and unchanged by the salinity in the medium. Addition of NaCl in the culturing solution increased the specific activity of the enzyme especially in the aerial parts, where GS is more abundant. This increase was all the more pronounced if the plant-salt contact period was extended (between 21 and 45 days). In vitro the addition of 0 to 500 m M of salt did not affect the activity of GS at satured substrate concentrations. At low glutamate concentrations in combination with 300 m M NaCl or more, a slight competitive inhibition was observed, never over 18%. – The remarkable insensibility of GS to salinity in vitro and the stimulating effect of NaCl in vivo on the synthesis of the leaf enzyme indicates that GS plays a fundamental part in the assimilation of NH4⁺ in the halophyte Suaeda maritima var. macrocarpa.     

Salinization of the soil solution decreases the further accumulation of salt in the root zone of the halophyte Atriplex nummularia Lindl. growing above shallow saline groundwater

Water use by plants in landscapes with shallow saline groundwater may lead to the accumulation of salt in the root zone. We examined the accumulation of Na⁺ and Cl^? around the roots of the halophyte Atriplex nummularia Lindl. and the impacts of this increasing salinity for stomatal conductance, water use and growth. Plants were grown in columns filled with a sand–clay mixture and connected at the bottom to reservoirs containing 20, 200 or 400 mM NaCl. At 21 d, Na⁺ and Cl^? concentrations in the soil solution were affected by the salinity of the groundwater, height above the water table and the root fresh mass density at various soil depths (P  < 0.001). However, by day 35, the groundwater salinity and height above the water table remained significant factors, but the root fresh mass density was no longer significant. Regression of data from the 200 and 400 mM NaCl treatments showed that the rate of Na⁺ accumulation in the soil increased until the Na⁺ concentration reached ~250 mM within the root zone; subsequent decreases in accumulation were associated with decreases in stomatal conductance. Salinization of the soil solution therefore had a feedback effect on further salinization within the root zone.     

硫腺苷甲硫氨酸合成酶基因的克隆及其在盐胁迫条件下的不同表达

硫腺苷甲硫氨酸作为甲基供体在转甲基反应中起到重要作用.为了解硫腺苷甲硫氨酸在盐地碱蓬(Suaedasalsa (L.)Pall)耐盐中的作用,我们对可能编码硫腺苷甲硫氨酸合成酶的基因(SsSAMS2)进行了分析.该基因在经400 mmol/L NaCl处理的盐地碱蓬地上部分的λ-Zap cDNA文库中克隆到,其插入片段全长1 531 bp,包含一个395个氨基酸的开放阅读框架,该基因推断的分子量约为43 kD.SsSAMS2与长春花(Catharanthus roseus)的SAMS2在氨基酸水平上的一致性为93%.Southern杂交显示,SsSAMS2在盐地碱蓬基因组中可能是两个拷贝.Northern分析显示硫腺苷甲硫氨酸合成酶基因受NaCl等胁迫的正调控.酶活性检测表明,NaCl胁迫条件下该酶活性增强.     

Properties of lead deposits in cell walls of radish (Raphanus sativus) roots

Various mechanisms are involved in detoxification of heavy metals such as lead (Pb) in plant cells. Most of the Pb taken up by plants accumulates in their roots. However, the detailed properties of Pb complexes in roots remain unclear. We have investigated the properties of Pb deposits in root cell walls of radish (Raphanus sativus L.) seedlings grown on glass beads bed containing Pb pellets, which are the source of Pb-contamination in shooting range soils. Pb deposits were tightly bound to cell walls. Cell wall fragments containing about 50,000 ppm Pb were prepared from the roots. After extracting Pb from the cell wall fragments using HCl, Pb ions were recombined with the Pb-extracted cell wall fragments in a solution containing Pb acetate. When the cell wall fragments were treated with pectinase (E.C. 3.2.1.15) and were chemically modified with 1-ethyl-3-dimethylamino-propylcarboimide, the Pb-rebinding ability of the treated cell wall fragments decreased. When acid-treated cell wall fragments were incubated in a solution containing Pb²⁺ and excess amounts of a chelating agent, Pb recombined with the cell wall fragments were measured to estimate the affinity between Pb²⁺ and the cell wall fragments. Our data show that Pb²⁺ binds to carboxyl groups of cell walls. The source of the carboxyl groups is suggested to be pectic compounds. A stability constant of the Pb-cell wall complex was estimated to be about 10⁸. The role of root cell walls in the mechanism underlying heavy metal tolerance was discussed.     

Caractérisation de la glutamate déshydrogénase chez un halophyte obligatoire: le Suaeda maritima var. macrocarpa

Characterization of the glutamate dehydrogenase from an obligate halophyte: Suaeda maritima var. macrocarpa The glutamate dehydrogenase (GDH), E.C. 1.4.1.3, of the obligate halophyte Suaeda maritima (L.) Dum. var. macrocarpa Moq. has been studied. NaCl increase in the culturing solution (0 to 23 g NaCl/litre) lowers the specific activity but does not affect the kinetic characteristics of the enzyme collected after purification on G-25 Sephadex gel. In optimal conditions, GDH is activated by the addition of 25 mM NaCl in the incubation medium and inhibited with concentrations over 100 mM. The inhibitory activity of the salt induces both a modification of the affinity of the enzyme for substrate (competitive inhibition for ketoglutarate) and a modification of the catalytic potency (non-competitive inhibition with NADH and ammonium ion). It is suggested that NaCl has a depressing influence on GDH synthesis. There is a fundamental difference between its short time action (in vitro) versus its continuous effect in the culturing solution.     

Effect of ABA on the contents of proline, polyamines, and cytokinins in the common ice plants under salt stress

The effects of ABA treatment on the contents of proline, polyamines (PA), and cytokinins (CK) in the facultative halophyte the common ice plant (Mesembryanthemum crystallinum L.) subjected to salt stress were studied. Plants grown in the phytotron chamber on Jonson nutrient medium for 6 weeks were subjected to 6-day-long salinity by a single NaCl adding to medium. During first three days of salinity, half plants of each treatment were placed for 30 min on nutrient medium containing 0, 100, or 300 mM NaCl plus ABA in the final concentration of 1 μM. Salinity reduced biomass accumulation and water and chlorophyll contents in plants. This was accompanied by the increase in the levels of MDA, proline, and sodium ions. ABA treatment of salt-stressed plants favored biomass accumulation and photosynthetic pigment protection, reduced the intensity of oxidative stress and the level of NaCl-induced proline accumulation. ABA treatment increased the contents of putrescine (Put) and spermidine (Spd) in the leaves and roots of control plants (not subjected to salt stress), reduced the losses of Put in the leaves and roots and Spd in the roots in the presence of 100 mM NaCl, and suppressed cadaverine (Cad) accumulation in the roots in the presence of 300 mM NaCl. In the presence of NaCl, ABA reduced the contents of zeatin and zeatin riboside and increased the level of zeatin-O-glucoside in the roots and isopentenyladenosine and isopentenyladenine in the leaves. Thus, ABA protective action under salinity can be realized through the weakening of oxidative stress (a decrease in MDA content) and the regulation of PA, proline, and CK metabolism, which has a great significance in plant adaptation to injurious factors.     

NaCl-Induced Alterations in Both Cell Structure and Tissue-Specific Plasma Membrane H+ -ATPase Gene Expression

NaCl-induced plasma membrane H+-ATPase gene expression, which occurs in roots and fully expanded leaves of the halophyte Atriplex nummularia L. (X. Niu, M.L. Narasimhan, R.A. Salzman, R.A. Bressan, P.M. Hasegawa [1993] Plant Physiol 103: 713-718), has been differentially localized to specific tissues using in situ RNA hybridization techniques. Twenty-four-hour exposure of plants to 400 mM NaCl resulted in substantial accumulation of H+ pump message in the epidermis of the root tip and the endodermis of the root elongation/differentiation zone. In expanded leaves, NaCl induction of plasma membrane H+-ATPase message accumulation was localized to bundle-sheath cells. Ultrastructural analyses indicated that significant cytological adaptations in root cells included plasmolysis that is accompanied by plasma membrane invaginations, formation of Hechtian strands and vesiculation, and vacuolation. These results identify specific tissues that are involved in the regulation of Na+ and Cl- uptake into different organs of the halophyte A. nummularia and provide evidence of the intercellular and interorgan coordination that occurs in the mediation of NaCl adaptation.     

Antioxidant responses of halophyte plant Aeluropus littoralis under long-term salinity stress

Salinity influences the agricultural production all over the world. This constrain, similar to others biotic and abiotic stresses generate the reactive oxygen species such as superoxide, hydrogen peroxide and hydroxyl radicals. In the evolution process of halophyte plants the mechanisms to detoxify ROS, such as antioxidant enzymes, have been developed. Aeluropus littoralis is a special halophyte that selected to our research, so the plants treated with NaCl at different salt concentration (0, 250, 450 and 650 mM) for a period 45 days. Leaves and roots (separately) collected and their proteins extracted for superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) and peroxidase (POD) activity assay. Meanwhile the electrolyte leakage of leaves analyzed and increased at 450 and 650 mM of NaCl concentrations. Superoxide dismutase and catalase showed same pattern for changing in enzymatic activities (increasing activity by salt stress in roots and decreasing in shoot at 450 and 650 mM stress), also peroxidase and ascorbate peroxidase activity almost increased in all stress conditions.     

Salt Tolerance in the Halophyte, Suaeda maritima (L.) Dum.: Properties of Malic Enzyme and PEP carboxylase

Malic enzyme and phosphenol pyruvate carboxylase activitieshave been isolated and characterized from the shoots of Suaedamaritima plants grown in culture solution (with and withoutNaCl) or in tap water. The enzymes isolated from the lattershowed increases in both specific activity and K_m values incomparison with plants grown in culture solution: however, theaddition of NaCl to the culture solution had no significanteffect on either enzyme. Malate levels were high in plants grownin tap water, reduced by an ordeT of magnitude by the additionof culture solution and then halved by the addition of NaCl. Both enzymes were inhibited in vitro by NaCl, although the additionof high concentrations of betaine and proline to the assay mediumdid not further inhibit enzyme activity. The significance ofthese results is discussed in relation to the proposed roleof betaine and proline as cytoplasmic osmoregulators. Suaeda maritima, halophyte, salt tolerance, malic enzyme, PEP carboxylase     

X-ray microanalysis of ion distribution within root cortical cells of the halophyte Suaeda maritima (L.) Dum.

The ion content of compartments within cortical cells of mature roots of the halophyte Suaeda maritima grown at 200 mol·m^-3 NaCl has been studied by X-ray microanalysis of freeze-substituted thin sections. Sodium and Cl were found in the vacuoles at about four-times the concentration in the cytoplasm or cell walls, whereas K was more concentrated in the cell walls and cytoplasm than in vacuoles. The vacuolar Na concentration was 12- to 13-times higher than that of K. The Na concentration of cell walls of cortical cells was about 95 mol·m^-3 of analysed volume. The cytoplasmic K concentration within the mature cortical cells was estimated to be 55 mol·m^-3 of analysed volume.