Different Effect of Humidity on Growth and Salt Tolerance of Two Soybean Cultivars

Two soybean (Glycine max (L.) Merr.) cultivars, Tachiyutaka and Dare, were grown in pots at 30 and 70 % relative humidity (RH) and treated with 0 (control), 40 (moderate), 80 and 120 (severe) mM NaCl for 3 weeks. Increasing RH enhanced growth of salt sensitive cultivar, Tachiyutaka, but had no effect on salt tolerant cultivar, Dare, under control and moderate saline conditions. Both cultivars benefited from elevated humidity under severe saline conditions. Cultivar Tachiyutaka had poorer ability for controlling translocation of Na⁺ to the leaves, lower Na⁺ exclusion ability in the roots, and lower root activity under NaCl treatment, compared with cv. Dare. The increased growth of cv. Tachiyutaka at high RH was consistent with decreased Na⁺ accumulation in the leaves, increased stomatal conductance and root activity, while the unchanged growth of cv. Dare was consistent with similar Na⁺ accumulation in the leaves, and the decreased root activity.     

Photosynthesis Parameters in Two Cultivars of Mulberry Differing in Salt Tolerance

Three-month-old mulberry (Morus alba L.) cultivars (salt tolerant cv. S1 and salt sensitive cv. ATP) were subjected to different concentrations of NaCl for 12 d. Leaf area, dry mass accumulation, total chlorophyll (Chl) content, net CO₂ assimilation rate (P _N), stomatal conductance (g _s), and transpiration rate (E) declined, and intercellular CO₂ concentration (C _i) increased. The changes in these parameters were dependent on stress severity and duration, and differed between the two cultivars. The tolerant cultivar showed a lesser reduction in P _N and g _s coupled with a better C _i and water use efficiency (WUE) than the sensitive cultivar. This revised version was published online in June 2006 with corrections to the Cover Date.     

Activity of Ion Transporters and Salt Tolerance in Barley

The authors attempted to relate the cultivar-specific salt tolerance in barley (Hordeum distichum L.) to the efficiency of ion transporters in the plasmalemma and tonoplast. The study involved plasmalemma and tonoplast membrane vesicles isolated from roots and leaves of the 7-day-old barley seedlings exposed to elevated NaCl concentrations. Two barley cultivars were employed: salt-tolerant cv. Elo and salt-susceptible cv. Belogorskii. The vesicles were used to measure the transport activity of plasmalemma and tonoplast proton pumps and the cation/anion exchange. The data obtained in the experiments demonstrated that the changes in the activity of ion transporters under salt stress conditions correlated with the barley cultivar-specific tolerance to elevated NaCl concentrations.     

Expression of a Novel Antiporter Gene from Brassica napus Resulted in Enhanced Salt Tolerance in Transgenic Tobacco Plants

Tobacco leaf discs were transformed with a plasmid pBIBnNHX1, containing the selectable marker neomycin phosphotransferase gene (nptII) and Na⁺/H⁺ vacuolar antiporter gene from Brassica napus (BnNHX1), via Agrobacterium tumefaciens-mediated transformation. Thirty-two independent transgenic plants were regenerated. Polymerase chain reaction (PCR) and Southern blot analyses confirmed that the BnNHX1 gene had integrated into plant genome and Northern blot analysis revealed the transgene expression at various levels in transgenic plants. Transgenic plants expressing BnNHX1 had enhanced salt tolerance and could grow and produce seeds normally in the presence of 200 mM NaCl. Analysis for the T₁ progenies derived from seven independent transgenic primary transformants expressing BnNHX1 showed that the transgenes in most tested independent T₁ lines were inherited at Mendelian 3:1 segregation ratios. Transgenic T₁ progenies could express _BnNHX1 and had salt tolerance at levels comparable to their T₀ parental lines. This study implicates that the BnNHX1 gene represents a promising candidate in the development of crops for enhanced salt tolerance by genetic engineering.     

Photosynthetic Parameters at the Vegetative Stage and during Grain Development of Two Hexaploid Wheat Cultivars Differing in Salt Tolerance

Response of two spring wheat (Triticum aestivum L.) cultivars, salt tolerant SARC-I and salt sensitive Potohar, to different concentrations of NaCl was examined under glasshouse conditions. Eighteen-day-old plants of both the lines grown in sand culture were irrigated with 0 (control), 80, 160 or 240 mM NaCl in full strength Hoagland's nutrient solution. Shoot fresh and dry masses, and leaf area per plant of SARC-I at the vegetative stage, were significantly greater than those of cv. Potohar at higher salt concentrations, however, relative growth rate (RGR) of cv. Potohar was significantly higher than that of SARC-I. SARC-I had higher net photosynthetic rate (P_N), stomatal conductance (g_s) and transpiration rate (E) than cv. Potohar at the vegetative stage, but the cultivars did not differ significantly in water-use efficiency (P_N/E), intrinsic water use efficiency (P_N/g_s), and intercellular/ambient CO₂ concentration ratio. At the grain development stage, SARC-I had significantly higher P_N and g_s in the flag leaf than cv. Potohar under salinity. SARC-I was superior to cv. Potohar with respect to number of grains per spike, number of grains per spikelet, mean grain mass, and grain yield per plant at all NaCl concentrations.     

Salt tolerance in rice in vitro: Implication of accumulation of Na+, K+ and proline

The implication of accumulation of both inorganic (Na⁺, K⁺) and organic (proline) solutes were evaluated in unadapted and NaCl-adapted callus of a salt-sensitive (Basmati 370) and a salt-tolerant (SR-26B) cultivar of rice (Oryza sativa L.) after a NaCl shock. Accumulation of Na⁺,K⁺ and/or proline in callus was co relatable and the relative presence of these components in tissues after shock treatment was found to be important factors to support differential regrowth capacities of the shock treated calluses. Presence or retention of K⁺ in rice callus was a key factor for salt tolerance as it was observed to be positively correlated with growth in both the varieties. The results indicated that K⁺ was the first candidate to counteract the negative water potential of outside milieu, while proline was probably the last metabolic device that rice calluses opted for when exposed to salt stress.     

Ionic and Osmotic Effects of Salinity on Single-Leaf Photosynthesis in Two Wheat Cultivars with Different Drought Tolerance

The effects of iso-osmotic salinity and drought stresses on leaf net photosynthetic rate (P _N) in two wheat (Triticum aestivum L.) cultivars BR 8 and Norin 61, differing in drought tolerance, were compared. In drought-sensitive Norin 61, the decline of P _N was larger than that in drought-tolerant BR 8. Under NaCl treatment, P _N decreased in two phases similarly in both cultivars. In the first phase, photosynthetic depression was gradual without any photochemical changes. In the second phase, photosynthetic depression was rapid and accompanied with a decline of the energy conversion efficiency in photosystem 2 (_PS2). Our observations suggest that the osmotic factor may induce a gradual depression of photosynthesis due to stomatal closure under both stress treatments. However, under NaCl treatment, a ionic factor (uptake and accumulation of excess Na⁺) may have direct effects on electron transport and cause more severe photosynthetic depression. The drought tolerance mechanism of BR 8 was insufficient to maintain single-leaf photosynthesis under salinity.     

Effects of Eu3+ on the metabolism of amino acid and protein in xerophytic Lathyrus sativus L.

The contents of amino acids and proteins and the activity of Na⁺, K⁺-ATPase were determined in roots, stems, and leaves of Eu³⁺-treated Lathyrus sativus L. The results showed that the treatment of Eu³⁺ made the contents of amino acid and protein and the activity of Na⁺, K⁺-ATPase change. The first possible mechanism was that Eu³⁺ directly made the electric potential of −NH₂ or −COOH of amino acid change. The second possible mechanism was that Eu³⁺ played a role in metallic-activated factors of certain enzymes, which catalyze the catabolism and anabolism of protein. Then, the contents of amino acids and proteins were relatively changed. The third possible mechanism was that Eu³⁺ regulated the activity of ATPase through changing the Na⁺/K⁺ ratio. The energy released by ATPase was the driving force for the translocation of amino acids and proteins in the plant cell. Because of the changeability of its valence, Eu played an, important role in regulating certain physiological reactions to increase the adaptability of L. sativus in arid environment. These authors contributed equally to this work.     

不同类型甜瓜成熟过程中果肉质地及其细胞显微结构的变化

采用质地剖面分析(TPA)和穿刺方法,测定3种不同口感质地的5个甜瓜材料(梗硬果肉的P10和3-6、脆酥果肉的417和20-5以及软果肉的Charentais)不同成熟时期果肉硬度、咀嚼性和黏着性以及脆性和平均硬度,评价甜瓜果肉的质地特性;采用组织切片法观察果肉组织细胞的显微结构,并通过Image-pro plus 6.0软件测定细胞大小参数(细胞面积、周长、长度及宽度)和形状参数(细胞纵横比及圆度),明确不同果肉质地类型甜瓜果实成熟过程中果肉细胞显微结构的变化特征,探讨甜瓜细胞形态参数与果肉质地的关系,为甜瓜品质育种提供理论依据。结果显示:(1)梗硬果肉甜瓜(P10和3-6)的果肉细胞较小,排列紧密;脆酥果肉甜瓜(417和20-5)的细胞较大,排列较疏松;软果肉甜瓜(Charentais)的细胞最大,排列极不规则。(2)不同口感甜瓜果肉在成熟期的细胞面积和周长差异显著,与口感呈显著负相关关系;在成熟过程中,甜瓜果肉细胞面积、周长和长度等表现出不同程度的增大,而细胞纵横比和圆度总体表现为下降趋势,即细胞越来越圆。(3)甜瓜果肉的口感与质地及细胞大小参数呈显著或极显著相关关系;细胞大小与黏着性呈显著或极显著正相关关系(0.951*~0.983**),细胞面积与TPA硬度及脆性呈显著负相关关系(分别为-0.910*和-0.926*),长度和宽度与脆性呈显著负相关关系(分别为-0.884*和-0.894*);细胞形状参数中的圆度与黏着性具有显著相关性(0.936*)。研究表明,口感不同的甜瓜果肉具有显著不同的质地和细胞显微结构,且甜瓜果肉口感与其果肉质地及细胞大小密切相关,即细胞越小,甜瓜果肉质地越硬。     

Effect of Osmotic Stress on Abscisic Acid Efflux and Compartmentation in the Roots of Two Maize Lines Differing in Drought Susceptibility

Roots of two Zea mays L. lines (drought-resistant Polj 17, and drought-susceptible F-2) were exposed to osmotic stress induced by sorbitol (osmotic potential –1.0 MPa). The following parameters were determined in cortex cells: membrane permeability for abscisic acid (ABA), ABA fluxes across membranes, pH values and ABA content in cytoplasm and vacuole. Osmotic stress induced different distribution of ABA within cell compartments in the investigated lines. ABA transport in the F-2 line occurred according to the intracellular pH gradient and the anion trap concept. In Polj 17, however, osmotic stress did not cause any significant effect on pH gradient and compartmental ABA content, but had a stimulating effect on ABA efflux from cytoplasm to apoplast and than via xylem to the leaf. These findings indicate different mechanisms of ABA transport between the investigated lines in response to osmotic stress.     

Short-Term Salinity Induced Changes in Two Wheat Cultivars at Different Growth Stages

Soluble sugars, proline, total chlorophyll contents and electrolyte leakage were measured in two wheat (Triticum aestivum L.) cultivars KRL 1-4 and HD 2009 at different growth stages [crown root initiation (CRI), flowering, and soft dough] under short term salinity (NaCl, CaCl₂ and Na₂SO₄). In control plants sugar contents were maximum at flowering stage. Proline and sugar concentrations increased in both cultivars under salinity with a maximum increase at CRI. Electrolyte leakage increased and chlorophyll content decreased with the plant age. A sharp increase of electrolyte leakage was noticed at salinity of 10 and 15 dS m^–1 in HD 2009 and KRL 1-4, respectively. The short-term salinity at CRI stage proved more detrimental as compared to salinity at flowering and soft dough stages in term of all biochemical changes induced. In wheat, plant resistance to salinity increased with the age of plant. The cultivar KRL 1-4 performed better under salinity as compared to HD 2009.     

Characteristics of Salt-Tolerant and Salt-Susceptible Cultivars of Barley

Twelve cultivars of barley (Hordeum vulgare L.) from the collection of the Vavilov Institute of Plant Industry, Russian Academy of Agricultural Sciences, were screened by assessing the length of 6-day-old seedlings grown in water culture at 70, 120, and 170 mM NaCl. As a result, two salt-susceptible cultivars, Belogorskii and QB 60.1, and three salt-tolerant cultivars, Elo, Odesskii 115, and Local from Ecuador, were selected, and these cultivars were used in the greenhouse soil-culture experiments. The grain yield of salt-tolerant cultivars was affected by NaCl to a lesser degree than that of the salt-susceptible cultivars. In both cases, soil salinization increased the sodium content in the seedlings as compared to the control plants. Characteristically, salt-susceptible cultivars accumulated more Na⁺ in their shoots than salt-tolerant cultivars; the reciprocal pattern was found in the roots. Soil salinization decreased K⁺ content in the shoots of the salt-susceptible cv. Belogorskii as compared to the control, whereas in the most tolerant cv. Local from Ecuador, the potassium content increased.     

Regulation of intracellular level of Na+, K+ and glycerol in Saccharomyces cerevisiae under osmotic stress

The intracellular level of Na⁺ and K⁺ of S. cerevisiae strain AB1375 revealed that under KCl as well as sorbitol stress, the cationic level was comparable to the level under no stress conditions. On the other hand, there was a sharp drop in the intracellular K⁺ content and increase in the Na⁺ content on addition of NaCl to the medium. However, the total cationic level was close to that under control conditions. In addition to changes in the cationic level, an enhanced production and accumulation of glycerol were also observed under osmotic stress. A regulatory mechanism co-ordinating the intracellular concentration of glycerol as well as Na⁺, K⁺ content under osmotic stress conditions has been proposed.     

The Effects of NaCl Treatment on Water Relations, Growth, and ABA Content in Barley Cultivars Differing in Drought Tolerance

Changes in transpiration and stomatal conductance and other characteristics of water relations, growth rate, and ABA content have been followed in short- and long-term experiments in two barley cultivars (cv. Michaelovsky and cv. Prairie) with contrasting drought resistance characteristics. The aim of this work was to reveal the importance of stomatal behavior in salt tolerance and also the involvement of ABA in its control. Salinity stress brought about a reduction in stomatal conductance in both cultivars, but the effect was initially more pronounced in the drought-tolerant cv. Prairie than in the drought-sensitive cv. Michaelovsky. The difference between the two cultivars changed with time, and later on transpiration and stomatal conductance became higher in Prairie than in Michaelovsky. In both the short and the long term, the extent of stomatal closure due to salinity correlated with the level of ABA accumulation in the leaves of the plants. Fast stomatal closure was likely to be responsible for growth resumption after an initial arrest by salt treatment and for the maintenance of extension growth later on, thus enabling its higher rate in Prairie than in Michaelovsky plants. Leaves of Prairie accumulated less toxic chloride ions, which may be the result of a lower transpiration rate observed during the first phase of salt treatment. A subsequent increase in stomatal conductance observed in Prairie is likely to ameliorate their gas exchange and maintain photosynthesis and growth. Thus, differences between the cultivars in the stomatal response to salinity changed with time, which may be why there are discrepancies in the attempts to relate stomatal conductance to salt tolerance observed in literature.     

Effects of Paclobutrazol on Response of Two Barley Cultivars to Salt Stress

The seeds of two barley (Hordeum vulgare L.) cultivars (a drought resistant cv. Tokak-137/57 and a drought sensitive cv. Erginel-90) were imbibed either in distilled water (control) or in a solution containing 40 mg dm⁻³ paclobutrazol (PBZ) and air dried. Seeds were germinated and grown in a glasshouse for 21 d and seedlings were subjected to salt stress by treating them with 100 and 200 mM NaCl for 12 d. The height of shoots was significantly decreased and root length was increased in PBZ-treated plants prior and after NaCl stress for 12 d leading to an increase in root to shoot ratio. Leaf chlorophyll and carotenoid contents in PBZ treated plants were increased in controls and especially in plants subjected to salt stress. PBZ induced increase in superoxide dismutase (SOD) activities was higher in cv. Tokak-157/37, than in cv. Erginel-90. However, an increase in SOD activity was not accompanied by an increase in peroxidase activity. This revised version was published online in July 2006 with corrections to the Cover Date.     

Na+/Na+ exchange and Na+/H+ antiport in rabbit erythrocytes: Two distinct transport systems

Summary Rabbit erythrocytes are well known for possessing highly active Na⁺/Na⁺ and Na⁺/H⁺ countertransport systems. Since these two transport systems share many similar properties, the possibility exists that they represent different transport modes of a single transport molecule. Therefore, we evaluated this hypothesis by measuring Na⁺ transport through these exchangers in acid-loaded cells. In addition, selective inhibitors of these transport systems such as ethylisopropyl-amiloride (EIPA) and N-ethylmaleimide (NEM) were used. Na⁺/Na⁺ exchange activity, determined as the Na _o ⁺ -dependent²²Na efflux or Na _i ⁺ -induced²²Na entry was completely abolished by NEM. This inhibitor, however, did not affect the H _i ⁺ -induced Na⁺ entry sensitive to amiloride (Na⁺/H⁺ exchange activity). Similarly, EIPA, a strong inhibitor of the Na⁺/H⁺ exchanger, did not inhibit Na⁺/Na^– countertransport, suggesting the independent nature of both transport systems. The possibility that the NEM-sensitive Na⁺/Na⁺ exchanger could be involved in Na⁺/H⁺ countertransport was suggested by studies in which the net Na⁺ transport sensitive to NEM was determined. As expected, net Na⁺ transport through this transport system was zero at different [Na⁺] _i /[Na⁺] _o ratios when intracellular pH was 7.2. However, at pH _i =6.1, net Na⁺ influx occurred when [Na⁺] _i was lower than 39mm. Valinomycin, which at low [K⁺] _o was lower than 39mm. Valinomycin, which at low [K⁺] _o clamps the membrane potential close to the K⁺ equilibrium potential, did not affect the net NEM-sensitive Na⁺ entry but markedly stimulated, the EIPA-and NEM-resistant Na⁺ uptake. This suggest that the net Na⁺ entry through the NEM-sensitive pathway at low pH _i , is mediated by an electroneutral process possibly involving Na⁺/H⁺ exchange. In contrast, the EIPA-sensitive Na⁺/H⁺ exchanger is not involved in Na⁺/Na⁺ countertransport, because Na⁺ transport through this mechanism is not affected by an increase in cell Na^– from 0.4 to 39mm. Altogether, these findings indicate that both transport systems: the Na⁺/Na⁺ and Na⁺/H⁺ exchangers, are mediated by distinct transport proteins.     

Root-pressure driven xylem sap flow in greenhouse melon (Cucumis melo L.): diurnal change and the effects of shading, growth stage, rootstock and fruit number

Grafted and ungrafted greenhouse melon were used to investigate the effect of diurnal change, shading, growth stage, rootstock and fruit numbers on melon xylem sap flow rate. A clear diurnal change was observed in xylem sap flow rate. Shading of the plant on the previous day decreased the sap flow rate. An increase in the number of fruit on a plant decreased the sap flow rate, but, grafting to squash plant lessened the effect of fruit number. Consideration of timing and environmental factors is necessary when the xylem sap is collected for root study.     

Salt tolerance conferred by overexpression of Arabidopsis vacuolar Na+/H+ antiporter gene AtNHX1 in common buckwheat (Fagopyrum esculentum)

Agriculture productivity is severely affected by soil salinity. One possible mechanism by which plants could survive salt stress is to compartmentalize sodium ions away from the cytosol. In the present work, transgenic buckwheat plants overexpressing AtNHX1, a vacuolar Na⁺/H⁺ antiporter gene from Arabidopsis thaliana, were regenerated after transformation with Agrobacterium tumefaciens. These plants were able to grow, flower and accumulate more rutin in the presence of 200 mmol/l sodium chloride. Moreover, the content of important nutrients in buckwheat was not affected by the high salinity of the soil. These results demonstrated the potential value of these transgenic plants for agriculture use in saline soil.     

Structural and Functional Analysis of Transmembrane Segment IV of the Salt Tolerance Protein Sod2

Sod2 is the plasma membrane Na⁺/H⁺ exchanger of the fission yeast Schizosaccharomyces pombe. It provides salt tolerance by removing excess intracellular sodium (or lithium) in exchange for protons. We examined the role of amino acid residues of transmembrane segment IV (TM IV) (¹²⁶FPQINFLGSLLIAGCITSTDPVLSALI¹⁵²) in activity by using alanine scanning mutagenesis and examining salt tolerance in sod2-deficient S. pombe. Two amino acids were critical for function. Mutations T144A and V147A resulted in defective proteins that did not confer salt tolerance when reintroduced into S. pombe. Sod2 protein with other alanine mutations in TM IV had little or no effect. T144D and T144K mutant proteins were inactive; however, a T144S protein was functional and provided lithium, but not sodium, tolerance and transport. Analysis of sensitivity to trypsin indicated that the mutations caused a conformational change in the Sod2 protein. We expressed and purified TM IV (amino acids 125–154). NMR analysis yielded a model with two helical regions (amino acids 128–142 and 147–154) separated by an unwound region (amino acids 143–146). Molecular modeling of the entire Sod2 protein suggested that TM IV has a structure similar to that deduced by NMR analysis and an overall structure similar to that of Escherichia coli NhaA. TM IV of Sod2 has similarities to TM V of the Zygosaccharomyces rouxii Na⁺/H⁺ exchanger and TM VI of isoform 1 of mammalian Na⁺/H⁺ exchanger. TM IV of Sod2 is critical to transport and may be involved in cation binding or conformational changes of the protein.