Salicylic Acid Induces Salinity Tolerance in Tomato (Lycopersicon esculentum cv. Roma): Associated Changes in Gas Exchange, Water Relations and Membrane Stabilisation

The present study investigates the role of salicylic acid (SA) in inducing plant tolerance to salinity. The application of 0.1 mM SA to tomato [Lycopersicon esculentum Mill.] plants via root drenching provided protection against 150 mM or 200 mM NaCl stress. SA treated plants had greater survival and relative shoot growth rate compared to untreated plants when exposed to salt stress. At 200 mM salt, shoot growth rates were approximately 4 times higher in SA treated plants than untreated plants. Application of SA increased photosynthetic rates in salt stressed plants and may have contributed to the enhanced survival. Transpiration rates and stomatal conductance were also significantly higher in SA treated plants under saline stress conditions. SA application reduced electrolyte leakage by 44% in 150 mM NaCl and 32% in 200 mM NaCl, compared to untreated plants, indicating possible protection of integrity of the cellular membrane. Beneficial effects of SA in saline conditions include sustaining the photosynthetic/transpiration activity and consequently growth, and may have contributed to the reduction or total avoidance of necrosis. SA, when used in appropriate concentrations, alleviates salinity stress without compromising the plants ability for growth under a favourable environment.     

Understanding Saline and Osmotic Tolerance of Populus euphratica Suspended Cells

Tolerance of Populus euphratica suspended cells to ionic and osmotic stresses implemented respectively by NaCl and PEG (6000) was characterized by monitoring cell growth, morphological features, ion compartmentation and polypeptide patterns. The cells grew and proliferated when submitted to stresses of 137 mM NaCl or 250 g l⁻¹ PEG, and survived at 308 mM of NaCl, showing tolerance to saline and particularly osmotic stress. They were resistant to plasmolysis and had dense cytoplasms, large nuclei and nucleoli, and evident cytoplasmic strands under high saline and osmotic stress. The sequestration of Cl⁻ into the vacuoles was observed in the cells stressed with 137 and 223 mM NaCl. The cellular protein profile was modified by high salt and osmotic stress and showed 28 kDa polypeptides up-regulated by both NaCl and PEG, and 66 and 25 kDa polypeptides up-regulated only by high NaCl stress. The salt tolerance of P. euphratica cells might be related to their capacity of adapting to higher osmotic stress by maintaining cell integrity, sequestrating Cl⁻ into vacuoles and modulating polypeptides that reflect cellular metabolic adaptations.     

Glucose-6-phosphate dehydrogenase acts as a regulator of cell redox balance in rice suspension cells under salt stress

The reduced coenzyme nicotinamide-adenine dinucleotide phosphate (NADPH) is an important molecule in cellular redox balance. Glucose-6-phosphate dehydrogenase (G6PDH) is a key enzyme in the pentose phosphate pathway, the most important NADPH-generating pathway. In this study, roles of G6PDH in maintaining cell redox balance in rice suspension cells under salt stress were investigated. Results showed that the G6PDH activity decreased in the presence of 80 mM NaCl on day 2. Application of exogenous glucose stimulated the activity of G6PDH and NADPH oxidase under salt stress. Exogenous glucose also increased the ion leakage, thiobarbituric acid reactive substances and hydrogen peroxide (H₂O₂) contents in the presence of 80 mM NaCl on day 2, implying that the reduction of the G6PDH activity was necessary to avoid serious damage caused by salt stress. The NAPDH/NADP⁺ ratio increased on day 2 but decreased on day 4 under 80 mM NaCl plus glucose treatment. Diphenyleneiodonium, an NADPH oxidase inhibitor, decreased the H₂O₂ content under 80 mM NaCl treatment on day 2. These results imply that the H₂O₂ accumulation induced by glucose treatment under salt stress on day 2 was related to the NADPH oxidase. Western-blot analysis showed that the G6PDH expression was slightly induced by glucose and was obviously blocked by DPI on day 2 under salt stress. In conclusion, G6PDH plays a key role in maintaining the cell redox balance in rice suspension cells under salt stress. The coordination of G6PDH and NADPH oxidase is required in maintaining cell redox balance in salt tolerance.     

Arabidopsis sos1 mutant in a salt-tolerant accession revealed an importance of salt acclimation ability in plant salt tolerance

An analysis of the salinity tolerance of 354 Arabidopsis thaliana accessions showed that some accessions were more tolerant to salt shock than the reference accession, Col-0, when transferred from 0 to 225 mM NaCl. In addition, several accessions, including Zu-0, showed marked acquired salt tolerance after exposure to moderate salt stress. It is likely therefore that Arabidopsis plants have at least two types of tolerance, salt shock tolerance and acquired salt tolerance. To evaluate a role of well-known salt shock tolerant gene SOS1 in acquired salt tolerance, we isolated a sos1 mutant from ion-beam-mutagenized Zu-0 seedlings. The mutant showed severe growth inhibition under salt shock stress owing to a single base deletion in the SOS1 gene and was even more salt sensitive than Col-0. Nevertheless, it was able to survive after acclimation on 100 mM NaCl for 7 d followed by 750 mM sorbitol for 20 d, whereas Col-0 became chlorotic under the same conditions. We propose that genes for salt acclimation ability are different from genes for salt shock tolerance and play an important role in the acquisition of salt or osmotic tolerance.     

Changes in growth, lipid peroxidation and some key antioxidant enzymes in chickpea genotypes under salt stress

The present study was conducted to evaluate the effect of NaCl on growth and some key antioxidants in chickpea. Eight genotypes of chickpea were grown hydroponically for 15 days and then treated with different concentrations of salt [0 mM (T0), 25 mM (T1), 50 mM (T2), 75 mM (T3), and 100 mM (T4)]. Salinity showed marked changes in growth parameters (fresh and dry weight of root and shoot). The level of lipid peroxidation was measured by estimating malondialdehyde content. Lipid peroxidation increases with the increase in NaCl concentration in all genotypes but salt-tolerant genotypes (SKUA-06 and SKUA-07) were least affected as compared to other genotypes. The chlorophyll content was also affected with elevated levels of NaCl. Increased concentration of salt increased the activity of antioxidant enzymes like superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) and glutathione reductase in all chickpea genotypes but maximum activity was observed in salt-tolerant (SKUA-06 and SKUA-07) genotypes. Two genotypes of salt-tolerant and salt-sensitive varieties were analyzed further by real time PCR which revealed that the expression of SOD, APX and CAT genes were increased by NaCl in the salt-tolerant variety. The enhancement in tolerance against salt stress indicates that the genes involved in the antioxidative process are triggered by oxidative stress induced by environmental change. The results indicate that NaCl-induced oxidative stress hampers the normal functioning of the cell. The efficient antioxidants play a great role in mitigating the effect of NaCl stress in chickpea. This screening of NaCl-tolerant genotypes of chickpea can be performed on salt-affected land.     

The putative plasma membrane Na(+)/H(+) antiporter SOS1 controls long-distance Na(+) transport in plants

The salt tolerance locus SOS1 from Arabidopsis has been shown to encode a putative plasma membrane Na(+)/H(+) antiporter. In this study, we examined the tissue-specific pattern of gene expression as well as the Na(+) transport activity and subcellular localization of SOS1. When expressed in a yeast mutant deficient in endogenous Na(+) transporters, SOS1 was able to reduce Na(+) accumulation and improve salt tolerance of the mutant cells. Confocal imaging of a SOS1-green fluorescent protein fusion protein in transgenic Arabidopsis plants indicated that SOS1 is localized in the plasma membrane. Analysis of SOS1 promoter-beta-glucuronidase transgenic Arabidopsis plants revealed preferential expression of SOS1 in epidermal cells at the root tip and in parenchyma cells at the xylem/symplast boundary of roots, stems, and leaves. Under mild salt stress (25 mM NaCl), sos1 mutant shoot accumulated less Na(+) than did the wild-type shoot. However, under severe salt stress (100 mM NaCl), sos1 mutant plants accumulated more Na(+) than did the wild type. There also was greater Na(+) content in the xylem sap of sos1 mutant plants exposed to 100 mM NaCl. These results suggest that SOS1 is critical for controlling long-distance Na(+) transport from root to shoot. We present a model in which SOS1 functions in retrieving Na(+) from the xylem stream under severe salt stress, whereas under mild salt stress it may function in loading Na(+) into the xylem.     

Effect of salt stress on growth parameters, enzymatic antioxidant system, and lipid peroxidation in wild chicory (Cichorium intybus L.)

Efficient utilization of saline land for food cultivation can increase agricultural productivity and rural income. To obtain information on the salt tolerance/susceptibility of wild chicory (Cichorium intybus L.), the influence of salinity (0–260 mM NaCl) on chicory seed germination and that of two salinity levels of irrigation water (100 and 200 mM NaCl) on plant growth, antioxidative enzyme activity, and accumulation of proline and malondialdehyde (MDA) were investigated. The trials were performed outdoors, in pots placed under a protective glass covering, for two consecutive years. Seeds showed a high capacity to germinate in saline conditions. The use of 100 mM NaCl solution resulted in 81 % germination, whereas seed germinability decreased below 40 % using salt concentrations above 200 mM NaCl. Wild chicory showed tolerance to medium salinity (100 mM NaCl), whereas a drastic reduction in biomass was observed when 200 mM NaCl solution was used for irrigation. MDA, present in higher amounts in leaves than in roots, decreased in both tissues under increasing salinity. Proline content increased remarkably with the level of salt stress, more so in roots than in leaves. In salt stress conditions, the activity of antioxidant enzymes (APX, CAT, POD, SOD) was enhanced. The electrophoretic patterns of the studied enzymes showed that the salinity of irrigation water affected only the intensity of bands, but did not activate new isoforms. Our results suggest that wild chicory is able to grow in soil with moderate salinity by activating antioxidative responses both in roots and leaves.     

Allene Oxide Cyclase from <Emphasis Type="Italic">Camptotheca acuminata</Emphasis> Improves Tolerance Against Low Temperature and Salt Stress in Tobacco and Bacteria

Allene oxide cyclase (AOC, E 5.3.99.6) is an essential enzyme in jasmonate (JA) biosynthetic pathway. An AOC gene (defined as CaAOC, Database Accession No. AY863428) had been isolated from Camptotheca acuminata in previous work. Real-time quantitative PCR analysis indicated that mRNA expression of CaAOC was induced by salt stress (120 mM NaCl) and low temperature (4 degrees C). In order to further investigate the role of AOC gene in the processes, CaAOC was introduced into tobacco via Agrobacterium tumefaciens, and the transgenic lines were subjected to the examination of tolerance against salt stress and low temperature. Under salt stress, the chlorophyll content in transgenic tobacco was higher than that of in the wild plants. The electrolyte leakage test revealed that transgenic tobacco plants were more resistant to low temperature over control. Furthermore, 5'-truncated CaAOC was inserted into pET30 and then expressed in Escherichia coli strain BL21DE3 (pLysS). Interestingly, the transformants could grow on 2YT agar containing 400 mM NaCl. Although these mechanisms are not clear yet, this study suggested that CaAOC could not only be a potential target gene in the engineering of plants and bacteria for improved endurance against salt stress, but also be quite useful in enhancing plant tolerance to cold.     

Role of Salicylic Acid in Promoting Salt Stress Tolerance and Enhanced Artemisinin Production in Artemisia annua L.

In the present investigation, the role of salicylic acid (SA) in inducing salinity tolerance was studied in Artemisia annua L., which is a major source of the antimalarial drug artemisinin. SA, when applied at 1.00 mM, provided considerable protection against salt stress imposed by adding 50, 100, or 200 mM NaCl to soil. Salt stress negatively affected plant growth as assessed by length and dry weight of shoots and roots. Salinity also reduced the values of photosynthetic attributes and total chlorophyll content and inhibited the activities of nitrate reductase and carbonic anhydrase. Furthermore, salt stress significantly increased electrolyte leakage and proline content. Salt stress also induced oxidative stress as indicated by the elevated levels of lipid peroxidation compared to the control. A foliar spray of SA at 1.00 mM promoted the growth of plants, independent of salinity level. The activity of antioxidant enzymes, namely, catalase, peroxidase, and superoxide dismutase, was upregulated by salt stress and was further enhanced by SA treatment. Artemisinin content increased at 50 and 100 mM NaCl but decreased at 200 mM NaCl. The application of SA further enhanced artemisinin content when applied with 50 and 100 mM NaCl by 18.3 and 52.4%, respectively. These results indicate that moderate saline conditions can be exploited to obtain higher artemisinin content in A. annua plants, whereas the application of SA can be used to protect plant growth and induce its antioxidant defense system under salt stress.     

Physiological and Biochemical Characteristics and Response Patterns of Salinity Stress Responsive Genes (SSRGs) in Wild Quinoa (<i>Chenopodium quinoa</i> L.)

Cultivating salt-tolerant crops is a feasible way to effectively utilize saline-alkali land and solve the problem of underutilization of saline soils. Quinoa, a protein-comprehensive cereal in the plant kingdom, is an exceptional crop in terms of salt stress tolerance level. It seems an excellent model for the exploration of salt-tolerance mechanisms and cultivation of salt-tolerant germplasms. In this study, the seeds and seedlings of the quinoa cultivar Shelly were treated with different concentrations of NaCl solution. The physiological, biochemical characteristics and agronomic traits were investigated, and the response patterns of three salt stress-responsive genes (SSRGs) in quinoa were determined by real-time PCR. The optimum level of stress tolerance of quinoa cultivar Shelly was found in the range of 250–350 mM concentration of NaCl. Salt stress significantly induced expression of superoxide dismutase (SOD), peroxidase (POD), and particularly betaine aldehyde dehydrogenase (BADH). BADH was discovered to be more sensitive to salt stress and played an important role in the salt stress tolerance of quinoa seedlings, particularly at high NaCl concentrations, as it displayed upregulation until 24 h under 100 mM salt treatment. Moreover, it showed upregulation until 12 h under 250 mM salt stress. Taken together, these results suggest that BADH played an essential role in the salt-tolerance mechanism of quinoa. Based on the expression level and prompt response induced by NaCl, we suggest that the BADH can be considered as a molecular marker for screening salt-tolerant quinoa germplasm at the early stages of crop development. Salt treatment at different plant ontogeny or at different concentrations had a significant impact on quinoa growth. Therefore, an appropriate treatment approach needs to be chosen rationally in the process of screening salt-tolerant quinoa germplasm, which is useful to the utilization of saline soils. Our study provides a fundamental information to deepen knowledge of the salt tolerance mechanism of quinoa for the development of salt-tolerant germplasm in crop breeding programs.     

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.     

Behaviour of antioxidant defense system in the adaptive response to salt stress in Helianthus annuus L. cells

A relationship between the antioxidant defense system and salt tolerance in two types of sunflower calli differing in salt sensitivity was studied. No reduction in growth occurred in the NaCl-salt-adapted cell line (T) when grown on 175 mM NaCl but growth of the salt-stressed cell line (S) was reduced by 83%. Lipid peroxidation and protein oxidation increased during acute stress of salt stressed cells at 14 and 28 d of the experiment, while salt-adapted calli (T) remained similar to non-shocked (C) values. The antioxidant defense system of callus adapted to growth under NaCl responded differently to 175 mM of salt compared with the corresponding controls under shock treatment. Salt-adapted and salt-stressed calli showed a similar pattern in GSH content at day 14 but at day 28 in S calli, GSH content was increased 100% over the non-shocked calli, while T calli returned to the initial values. In the salt-stressed calli, a general decrease in all the antioxidant enzymes studied (except for glutathione reductase and dehydroascorbate reductase activities) was observed at day 28. Except for catalase, the antioxidant enzymes were elevated constitutively in adapted calli as compared to stressed cells, when both were grown in the absence of NaCl (time 0), and remained unaltered until 28 d after the beginning of the experiment. These results suggest the involvement of an enzymatic antioxidant defense system in the adaptive response to salt stress in Helianthus annuus L. cells.     

Messenger RNA induction in cellular salt tolerance of Alfalfa (Medicago sativa)

A salt tolerant alfalfaMedicago sativa L. cell line (HG2-N1) has been selected for growth in 171 mM NaCl. The salt tolerance characteristic is stable and is retained after growth in absence of salt selection for two months.In vitro translation was used to compare mRNA composition from the salt tolerant HG2-N1 and parent salt sensitive HG2 cell lines grown in the presence and absence of 171 mM NaCl. The results suggest that the mRNA composition differs between HG2-N1 and HG2 in a number of RNA species. The salt tolerant HG2-N1 shows both increases and decreases in specific polypeptides as compared to HG2. Many of the enhanced polypeptide bands from mRNA in the salt tolerant HG2-N1 variant appear to be constitutively expressed, since they can be detected from HG2-N1 cells grown in presence and absence of NaCl, but the expression of a few bands may depend on the presence of added NaCl. Most enhanced polypeptides, which are detected from mRNA in the salt tolerant variant HG2-N1 (grown on NaCl) are different from polypeptide bands enhanced in the salt sensitive HG2 line as a result of 24 hour salt stress. Similar results were obtained from two dimensional analysis ofin vivo labeled polypeptides. At least one isolated cDNA clone shows selective expression of mRNA in salt tolerant cells grown in NaCl. These results indicate that adaptive mechanisms for salt tolerance may differ in some aspects from acute stress mechanisms.     

Effect of salt stress on antioxidant defense systems,lipid peroxidation,and chlorophyll content in green bean

Salt stress-induced changes in antioxidant enzymes, such as catalase (CAT), ascorbate peroxidase (APX), and glutathione reductase (GR), total chlorophyll content, and lipid peroxidation measured as malondialdehyde (MDA) content, in leaves of a green bean genotype Gevas sirsk 57 (GS57) and cv. Fransiz 4F-89 differing in salt tolerance were investigated. Plants were subjected to three salt treatments (0, 50, and 100 mM NaCl) under controlled climatic conditions for 7 days. The salt-sensitive cv. 4F-89 exhibited a decrease in GR activity at all salt treatments, but the salt-tolerant genotype GS57 showed only a slight decrease in GR under 50 mM salt treatment and an increase under 100 mM salt treatment. CAT and APX activities increased with increasing salt stress in both varieties. CAT and APX activities were higher in the salt-tolerant GS57 than salt-ensitive cv. 4F-89. The two varieties showed an increase in MDA content with an increase in salinity, but the increase in sensitive cv. 4F-89 under salt stress was higher than that in salt-tolerant GS57 genotype. The increasing NaCl concentration caused a reduction in the chlorophyll content in cv. 4F-89 but not in GS57.     

In vitro recurrent selection of potato: production and characterization of salt tolerant cell lines and plants

A stable salt-tolerant potato cell line, able to grow on media containing 60–450 mM NaCl (i.e. low to high salinity) was selected. Callus grown on 120 or 150 mM NaCl showed higher fresh weights than the rest of the treatments. Replacing NaCl by KCl or Na2SO4 showed that reductions in fresh weight were mainly due to the presence of Na+ ions. When PEG 6000 was added to the medium instead of salt, the salt tolerant cell lines were unable to overcome the PEG-induced water stress. Whole plants, regenerated from salt tolerant callus, exhibited salt stress tolerance as evidenced by their higher fresh and dry weights when watered with 90 mM NaCl, and they also produced more tubers per plant under salt stress. Salt-tolerant plants differed phenotypically from control plants both in terms of leaf shape, tuber flesh and skin colour, which was reddish. In addition, DNA fingerprinting by RAPDs, with 70 different primers, confirmed that the salt tolerant regenerants also differed genotypically from the control, salt sensitive Kennebec potato plants from which they had been selected. This revised version was published online in June 2006 with corrections to the Cover Date.     

Physiological and epigenetic analyses of <Emphasis Type="Italic">Brassica napus</Emphasis> seed germination in response to salt stress

Salinity stress significantly affects plant growth and development because of osmotic stress, ion toxicity, and nutrient imbalance. Therefore, salinity stress becomes a serious threat to rapeseed production in agriculture. Plants evolved a series of complex mechanisms, including morphological changes, physiological adjustment, and gene expression regulation, at a molecular level to adapt to salt stress. Epigenetic regulations, including DNA methylation and histone modification, play a major role in tuning gene expression in plant response to environmental stimuli. Although many progresses have been reported in plant response to salt stress, the epigenetic changes in Brassica napus under salt stress are far from being understood. A series of physiological parameters, including water content, proline content, malondialdehyde content, electrolyte leakage, and antioxidant enzyme activities, under different concentrations (0, 25, 50, and 100 mM) of NaCl treatment in “Yangyou 9” was determined at the germination stage. Immunofluorescent staining and high-performance liquid chromatography-assisted quantification were conducted to analyze the level and distribution patterns of DNA and histone methylation under salt stress. Results of morphological and physiological analyses under salt stress indicated that 25 mM NaCl treatment promoted the growth of “Yangyou 9” seedlings, whereas 50 and 100 mM NaCl treatments inhibited the growth of “Yangyou 9” seedlings. Epigenetic investigations showed that 25 mM NaCl mediated the enrichment of H3K4me3, as well as decreases in H3K9me2 and 5-methylcytosine (5-mC), whereas 50 and 100 mM NaCl induced increases in H3K9me2 and 5-mC and a decrease in H3K4me3. Overall, this study offers new insights into the epigenetic changes in salt stress response in rapeseed, and this information would be propitious to engineer crops with enhanced salt tolerance.     

Salt tolerance, salt accumulation, and ionic homeostasis in an epidermal bladder-cell-less mutant of the common ice plant Mesembryanthemum crystallinum

The aerial surfaces of the common or crystalline ice plant Mesembryanthemum crystallinum L., a halophytic, facultative crassulacean acid metabolism species, are covered with specialized trichome cells called epidermal bladder cells (EBCs). EBCs are thought to serve as a peripheral salinity and/or water storage organ to improve survival under high salinity or water deficit stress conditions. However, the exact contribution of EBCs to salt tolerance in the ice plant remains poorly understood. An M. crystallinum mutant lacking EBCs was isolated from plant collections mutagenized by fast neutron irradiation. Light and electron microscopy revealed that mutant plants lacked EBCs on all surfaces of leaves and stems. Dry weight gain of aerial parts of the mutant was almost half that of wild-type plants after 3 weeks of growth at 400 mM NaCl. The EBC mutant also showed reduced leaf succulence and leaf and stem water contents compared with wild-type plants. Aerial tissues of wild-type plants had approximately 1.5-fold higher Na(+) and Cl(-) content than the mutant grown under 400 mM NaCl for 2 weeks. Na(+) and Cl(-) partitioning into EBCs of wild-type plants resulted in lower concentrations of these ions in photosynthetically active leaf tissues than in leaves of the EBC-less mutant, particularly under conditions of high salt stress. Potassium, nitrate, and phosphate ion content decreased with incorporation of NaCl into tissues in both the wild type and the mutant, but the ratios of Na(+)/K(+) and Cl(-)/NO(3)(-)content were maintained only in the leaf and stem tissues of wild-type plants. The EBC mutant showed significant impairment in plant productivity under salt stress as evaluated by seed pod and seed number and average seed weight. These results clearly show that EBCs contribute to succulence by serving as a water storage reservoir and to salt tolerance by maintaining ion sequestration and homeostasis within photosynthetically active tissues of M. crystallinum.     

Glucose-6-phosphate dehydrogenase plays a pivotal role in nitric oxide-involved defense against oxidative stress under salt stress in red kidney bean roots

The pivotal role of glucose-6-phosphate dehydrogenase (G-6-PDH)-mediated nitric oxide (NO) production in the tolerance to oxidative stress induced by 100 mM NaCl in red kidney bean (Phaseolus vulgaris) roots was investigated. The results show that the G-6-PDH activity was enhanced rapidly in the presence of NaCl and reached a maximum at 100 mM. Western blot analysis indicated that the increase of G-6-PDH activity in the red kidney bean roots under 100 mM NaCl was mainly due to the increased content of the G-6-PDH protein. NO production and nitrate reductase (NR) activity were also induced by 100 mM NaCl. The NO production was reduced by NaN(3) (an NR inhibitor), but not affected by N(omega)-nitro-L-arginine (L-NNA) (an NOS inhibitor). Application of 2.5 mM Na(3)PO(4), an inhibitor of G-6-PDH, blocked the increase of G-6-PDH and NR activity, as well as NO production in red kidney bean roots under 100 mM NaCl. The activities of antioxidant enzymes in red kidney bean roots increased in the presence of 100 mM NaCl or sodium nitroprusside (SNP), an NO donor. The increased activities of all antioxidant enzymes tested at 100 mM NaCl were completely inhibited by 2.5 mM Na(3)PO(4). Based on these results, we conclude that G-6-PDH plays a pivotal role in NR-dependent NO production, and in establishing tolerance of red kidney bean roots to salt stress.