Differential responses of antioxidative defense system to prolonged salinity stress in salt-tolerant and salt-sensitive Indica rice (Oryza sativa L.) seedlings

The present investigation evaluated the ability of an antioxidative defense system in terms of the tolerance against salinity-induced oxidative stress and also explored a possible relationship between the status of the components of an antioxidative defense system and the salt tolerance in Indica rice (Oryza sativa L.) genotypes. When the seedlings of a salt-sensitive cultivar was grown in sand cultures containing different NaCl concentrations (7 and 14 dS m^?1) for 5–20 days, a substantial increase was observed in the rate of superoxide anion (O ₂ ^·? ) production, elevated levels of H₂O₂ and thiobarbituric acid reactive substances (TBARS) which indicated an enhancement in lipid peroxidation. A declination in the level of thiol clearly indicated an increase in the protein oxidation as well as a decline in the reduced forms of ascorbate (AsA) and glutathione (GSH) and the ratios of their reduced to oxidized forms occurred in the salt-sensitive seedlings. Similar treatment caused a very little alteration or no change in the levels of these components in the seedlings of salt-tolerant cultivar. The activity of antioxidative enzymes superoxide dismutase (SOD), its isoform Cu/Zn-SOD and ascorbate peroxidase (APX) increased in both the cultivars against salinity. In salt-sensitive seedlings, the activity of the various enzymes, guaiacol peroxidase (GPX), catalase (CAT), monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR), and glutathione reductase (GR) increased at moderate salinity treatment of 7 dS m^?1 NaCl while the activities of these enzymes declined with higher salinity level of 14 dS m^?1 NaCl. However, a consistent increase was observed in the activities of these enzymes of salt-tolerant seedlings with an increase in the duration and the level of the salinity treatment. The results suggest that a higher status of antioxidants (AsA and GSH) and a coordinated higher activity of the enzymes (SOD, CAT, GPX, APX, and GR) can serve as the major determinants in the model for depicting salt tolerance in Indica rice seedlings.     

Comparative lipid peroxidation,antioxidant defense systems and proline content in roots of two rice cultivars differing in salt tolerance

The changes in the activity of antioxidant enzymes such as superoxide dismutase (SOD: EC 1.15.1.1), catalase (CAT: EC 1.11.1.6), peroxidase (POX: EC 1.11.1.7), ascorbate peroxidase (APOX: EC 1.11.1.11) and glutathione reductase (GR: EC 1.6.4.2), free proline content, and the rate of lipid peroxidation level in terms of malondialdehyde (MDA) in roots of two rice cultivars (cvs.) differing in salt tolerance were investigated. Plants were subjected to three salt treatments, 0, 60, and 120 mol m⁻³ NaCl for 7 days. The results showed that activated oxygen species may play a role in cellular toxicity of NaCl and indicated differences in activation of antioxidant defense systems between the two cvs. The roots of both cultivars showed a decrease in GR activity with increase in salinity. CAT and APOX activities increased with increasing salt stress in roots of salt-tolerant cultivar Pokkali but decreased and showed no change, respectively, in roots of IR-28 cultivar. POX activity decreased with increasing NaCl concentrations in salt-tolerant Pokkali but increased in IR-28. SOD activity showed no change in roots of both cultivars under increasing salinity. MDA level in the roots increased under salt stress in sensitive IR-28 but showed no change in Pokkali. IR-28 produced higher amount of proline under salt stress than in Pokkali. Increasing NaCl concentration caused a reduction in root fresh weight of Pokkali and root dry weight of IR-28. The results indicate that improved tolerance to salt stress in root tissues of rice plants may be accomplished by increased capacity of antioxidative system.     

Involvement of oxidative stress and role of antioxidative defense system in growing rice seedlings exposed to toxic concentrations of aluminum

When seedlings of rice (Oryza sativa L.) cultivar Pant-12 were raised in sand cultures containing 80 and 160 μM Al³⁺ in the medium for 5–20 days, a regular increase in Al³⁺ uptake with a concomitant decrease in the length of roots as well as shoots was observed. Al³⁺ treatment of 160 μM resulted in increased generation of superoxide anion (O₂ ⁻) and hydrogen peroxide (H₂O₂), elevated amount of malondialdehyde, soluble protein and oxidized glutathione and decline in the concentrations of thiols (-SH) and ascorbic acid. Among antioxidative enzymes, activities of superoxide dismutase (SOD EC 1.15.1.1), guaiacol peroxidase (Guaiacol POX EC 1.11.1.7), ascorbate peroxidase (APX EC 1.11.1.11), monodehydroascorbate reductase (MDHAR EC 1.6.5.4), dehydroascorbate reductase (EC 1.8.5.1) and glutathione reductase (EC 1.6.4.2) increased significantly, whereas the activities of catalase (EC EC 1.11.1.6) and chloroplastic APX declined in 160 μM Al³⁺ stressed seedlings as compared to control seedlings. The results suggest that Al³⁺ toxicity is associated with induction of oxidative stress in rice plants and among antioxidative enzymes SOD, Guaiacol POX and cytosolic APX appear to serve as important components of an antioxidative defense mechanism under Al³⁺ toxicity. PAGE analysis confirmed the increased activity as well as appearance of new isoenzymes of APX in Al³⁺ stressed seedlings. Immunoblot analysis revealed that changes in the activities of APX are due to changes in the amounts of enzyme protein. Similar findings were obtained when the experiments were repeated using another popular rice cv. Malviya-36.     

Reactive oxygen species, ascorbate–glutathione pool, and enzymes of their metabolism in drought-sensitive and tolerant indica rice (Oryza sativa L.) seedlings subjected to progressing levels of water deficit

Water deficit for rice is a worldwide concern, and to produce drought-tolerant varieties, it is essential to elucidate molecular mechanisms associated with water deficit tolerance. In the present study, we investigated the differential responses of nonenzymatic antioxidants ascorbate (AsA), glutathione (GSH), and their redox pool as well as activity levels of enzymes of ascorbate–glutathione cycle in seedlings of drought-sensitive rice (Oryza sativa L.) cv. Malviya-36 and drought-tolerant cv. Brown Gora subjected to water deficit treatment of ?1.0 and ?2.1 MPa for 24–72 h using PEG-6000 in sand cultures. Water deficit caused increased production of reactive oxygen species such as O₂?^?, H₂O₂, and HO? in the tissues, and the level of production was higher in the sensitive than the tolerant cultivar. Water deficit caused reduction in AsA and GSH and decline in their redox ratios (AsA/DHA and GSH/GSSG) with lesser decline in tolerant than the sensitive seedlings. With progressive level of water deficit, the activities of monodehydroascorbate reductase, dehydroascorbate reductase, ascorbate peroxidase (APX), and glutathione transferase increased in the seedlings of both rice cultivars, but the increased activity levels were higher in the seedlings of drought-tolerant cv. Brown Gora compared to the sensitive cv. Malviya-36. Greater accumulation of proline was observed in stressed seedlings of tolerant than the sensitive cultivar. In-gel activity staining of APX revealed varying numbers of their isoforms and their differential expression in sensitive and tolerant seedlings under water deficit. Results suggest that an enhanced oxidative stress tolerance by a well-coordinated cellular redox state of ascorbate and glutathione in reduced forms and induction of antioxidant defense system by elevated activity levels of enzymes of ascorbate–glutathione cycle is associated with water deficit tolerance in rice.     

Effects of 28-homobrassinolide on growth, lipid peroxidation and antioxidative enzyme activities in seedlings of Zea mays L. under salinity stress

The effects of 28-homobrassinolide (28-homoBL) on seedling growth, lipid peroxidation and antioxidative enzyme activities in the seedlings of Zea mays L. (var. Partap-1) under salt (NaCl) stress were studied. The surface-sterilized seeds were germinated in petriplates containing different concentrations of NaCl (25, 50, 75 and 100 mM) only, 28-homoBL (10⁻⁷, 10⁻⁹ and 10⁻¹¹ M) only and NaCl supplemented with 28-homoBL for 7 days. The activities of superoxide dismutase (SOD, EC 1.15.1.1), guaiacol peroxidase (POD, EC 1.11.1.7), catalase (CAT, EC 1.11.1.6), ascorbate peroxidase (APOX, EC 1.11.1.11) and glutathione reductase (EC 1.6.4.2) were analysed in 7 day-old seedlings. It was observed that 28-homoBL treatments reduced the toxicity of salt on seedling growth considerably. Lipid peroxidation level was significantly increased under saline stress, but lowered with 28-homoBL applications revealing less oxidative damage. Further 28-homoBL treatments to the seedlings showed an enhancement in activities of SOD, POD, CAT and APOX. The activities of all antioxidative enzymes were further increased in seedlings treated with solution containing 28-homoBL and salt together as compared to seedlings treated with different concentration of salt solution only.     

Effects of 24-epibrassinolide on seed germination, seedling growth, lipid peroxidation, proline content and antioxidative system of rice (Oryza sativa L.) under salinity stress

The effects of 24-epibrassinolide (24-epiBL) on seedling growth, antioxidative system, lipid peroxidation, proline and soluble protein content were investigated in seedlings of the salt-sensitive rice cultivar IR-28. Seedling growth of rice plants was improved by 24-epiBL treatment under salt stress conditions. When seedlings treated with 24-epiBL were subjected to 120 mM NaCl stress, the activities of superoxide dismutase (EC 1.15.1.1), catalase (EC 1.11.1.6) and glutathione reductase (EC 1.6.4.2) did not show significant difference, whereas the activity of ascorbate peroxidase (EC 1.11.1.11) significantly increased. Increased activity of peroxidase (EC 1.11.1.7) under NaCl stress showed remarkable decrease in the 24-epiBL+NaCl-applied group. Lipid peroxidation level significantly increased under salt stress but decreased with 24-epiBL application revealing that less oxidative damage occurred in this group (24-epiBL+NaCl). In addition, increased proline content in the NaCl-applied group was decreased by 24-epiBL application in the 24-epiBL+NaCl-applied group. Soluble protein content was increased by 24-epiBL application even under NaCl stress, being also higher than control conditions (no 24-epiBL or NaCl treatment). 24-epiBL treatment considerably alleviated oxidative damage that occurred under NaCl-stressed conditions and improved seedling growth in part under salt stress in sensitive IR-28 seedlings.     

Manganese-excess induces oxidative stress,lowers the pool of antioxidants and elevates activities of key antioxidative enzymes in rice seedlings

Manganese (Mn) is an essential element for plant growth but in excess, specially in acidic soils, it can become phytotoxic. In order to investigate whether oxidative stress is associated with the expression of Mn toxicity during early seedling establishment of rice plants, we examined the changes in the level of reactive oxygen species (ROS), oxidative stress induced an alteration in the level of non-enzymic antioxidants and activities of antioxidative enzymes in rice seedlings grown in sand cultures containing 3 and 6 mM MnCl₂. Mn treatment inhibited growth of rice seedlings, the metal increasingly accumulated in roots and shoots and caused damage to membranes. Mn treated plants showed increased generation of superoxide anion (O₂ ^.−), elevated levels of H₂O₂ and thiobarbituric acid reactive substances (TBARS) and decline in protein thiol. The level of nonprotein thiol, however, increased due to Mn treatment. A decline in contents of reduced ascorbate (AsA) and glutathione (GSH) as well as decline in ratios of their reduced to oxidize forms was observed in Mn-treated seedlings. The activities of antioxidative enzymes superoxide dismutase (SOD) and its isoforms Mn SOD, Cu/Zn SOD, Fe SOD as well as guaiacol peroxidase (GPX) increased in the seedlings due to Mn treatment however, catalase (CAT) activity increased in 10 days old seedlings but it declined by 20 days under Mn treatment. The enzymes of Halliwell-Asada cycle, ascorbate peroxidase (APX) monodehydoascorbate reductase (MDHAR), dehyroascorbate reductase (DHAR) and glutathione reductase (GR) increased significantly in Mn treated seedlings over controls. Results suggest that in rice seedlings excess Mn induces oxidative stress, imbalances the levels of antioxidants and the antioxidative enzymes SOD, GPX, APX and GR appear to play an important role in scavenging ROS and withstanding oxidative stress induced by Mn.     

Paclobutrazol mitigates salt stress in <Emphasis Type="Italic">indica</Emphasis> rice seedlings by enhancing glutathione metabolism and glyoxalase system

Stress-induced methylglyoxal (MG) functions as a toxic molecule, inhibiting plant physiological processes such as photosynthesis and antioxidant defense systems. In the present study, an attempt was made to investigate the MG detoxification through glutathione metabolism in indica rice [Oryza sativa L. ssp. indica cv. Pathumthani 1] under salt stress by exogenous foliar application of paclobutrazol (PBZ). Fourteen-day-old rice seedlings were pretreated with 15 mg L^?1 PBZ foliar spray. After 7 days, rice seedlings were subsequently exposed to 0 (control) or 150 mM NaCl (salt stress) for 12 days. Prolonged salt stress enhanced the production of MG molecules and the oxidation of proteins, leading to decreased activity of glyoxalase enzymes, glyoxalase I (Gly I) and glyoxalase II (Gly II). Consequently, the decreased glyoxalase activities were also associated with a decline in reduced glutathione (GSH) content and glutathione reductase (GR) activity. PBZ pretreatment of rice seedlings under salt stress significantly lowered MG production and protein oxidation, and increased the activities of both Gly I and Gly II. PBZ also increased GSH content and GR activity along with the up-regulation of glyoxalase enzymes, under salt stress. In summary, salinity induced a high level of MG and the associated oxidative damage, while PBZ application reduced the MG toxicity by up-regulating glyoxalase and glutathione defense system in rice seedlings.     

Salinity induced oxidative stress and antioxidant system in salt-tolerant and salt-sensitive cultivars of rice (Oryza sativa L.)

Indices of oxidative stress viz., superoxide radical and H₂O₂ content increased in leaves of all the cultivars with the rise in salinity level, the increase was more pronounced and significant in salt-sensitive varieties and non-significant in resistant cultivars. Except for glutathione reductase (GR), basal activities of all other antioxidative enzymes viz. superoxide dismutase (SOD), catalase (CAT), peroxidase (POX), ascorbate peroxidase (APX) and glutathione reductase (GR) were significantly higher in leaves of all the resistant cultivars as compared to the sensitive ones. A differential response of salinity was observed on various enzymatic and non-enzymatic components of antioxidant system in leaves of salt-tolerant and salt-sensitive cultivars of rice (Oryza sativa L.). Activities of superoxide dismutase and glutathione reductase enhanced in all the tolerant cultivar while declined in the sensitive cultivars with increasing salinity from 0 to 100 mM. Salt-stress induced the activities of catalase and peroxidase in all the cultivars but the magnitude of increase was more pronounced in the sensitive cultivars than in the tolerant cultivars. Contrarily, APX activity increased in the salt-sensitive cultivars but showed no significant change in the salt-tolerant cultivars. The amount of ascorbic acid content, reduced glutathione (GSH), reduced/oxidized glutathione (GSSG) ratio was higher in leaves of the tolerant cultivars than that of the sensitive cultivars under saline conditions. It is inferred that leaves of salt-tolerant cultivars tend to attain greater capacity to perform reactions of antioxidative pathway under saline conditions to combat salinity-induced oxidative stress.     

Paraquat pre-treatment increases activities of antioxidant enzymes and reduces lipid peroxidation in salt-stressed cucumber leaves

To investigate whether paraquat (PQ) is involved in regulation of antioxidant enzymes and lipid peroxidation under short-term salt stress, and to elucidate the physiological mechanism of salt stress mitigated by PQ, a cucumber cultivar (cv. Chunguang no. 2) was exposed to 100 mM NaCl for 48 h after pre-treatment with 10 μM PQ for 1 h. When compared to the control, salt stress increased the levels of malonaldehyde (MDA), superoxide radical (O₂^·−) and hydrogen peroxide (H₂O₂) and the activities of antioxidant enzymes, such as superoxide dismutase (EC 1.15.1.1), ascorbate peroxidase (EC 1.11.1.11) and glutathione reductase (EC 1.6.4.2) in the cucumber leaves. Under salt conditions, PQ pre-treatment prevented oxidative stress as observed by the decreases in MDA, H₂O₂ and O₂^·− that correlated with the increase in antioxidant defenses. We propose that, at low concentrations, the PQ pre-treatment can reduce the salt-induced oxidative damage by increasing the antioxidative mechanisms in cucumber plants.     

Response of the cultivated tomato and its wild salt-tolerant relative Lycopersicon pennellii to salt-dependent oxidative stress: The root antioxidative system

The response of the antioxidant system to salt stress was studied in the roots of the cultivated tomato Lycopersicon esculentum Mill. cv. M82 (Lem) and its wild salt-tolerant relative L. pennellii (Corr.) D'Arcy accession Atico (Lpa). Roots of control and salt (100 m M NaCl)-stressed plants were sampled at various times after commencement of salinization. A gradual increase in the membrane lipid peroxidation in salt-stressed root of Lem was accompanied with decreased activities of the antioxidant enzymes: superoxide dismutase (SOD; EC 1.15.1.1), catalase (CAT; EC 1.11.1.6), ascorbate peroxidase (APX; EC 1.11.1.11) and decreased contents of the antioxidants ascorbate and glutathione and their redox states. In contrast, increased activities of the SOD, CAT, APX, monodehydroascorbate reductase (MDHAR; EC 1.6.5.4), and increased contents of the reduced forms of ascorbate and glutathione and their redox states were found in salt-stressed roots of Lpa, in which the level of membrane lipid peroxidation remained unchanged. It seems that the better protection of Lpa roots from salt-induced oxidative damage results, at least partially, from the increased activity of their antioxidative system.     

Spermidine pretreatment enhances heat tolerance in rice seedlings through modulating antioxidative and glyoxalase systems

This study was undertaken to investigate the possible involvement of the antioxidant defense and glyoxalase systems in protecting rice seedlings from heat-induced damage in the presence of spermidine (Spd). Hydroponically grown 14-day-old seedlings were subjected to foliar spray with Spd (1 mM, 24 h) prior to heat stress (42 °C, 48 h) followed by subsequent recovery (27 °C, 48 h). Lipoxygenase activity, malondialdehyde (MDA), hydrogen peroxide (H₂O₂) and proline (Pro) content increased significantly whereas fresh weight (FW) and chlorophyll (Chl) content decreased during heat stress and after recovery, indicating unrecoverable damage to rice seedlings. Heat-induced damage was also evident in decreased levels of ascorbate (AsA), glutathione (GSH), and AsA and GSH redox ratios. Superoxide dismutase (SOD) and catalase (CAT) activities increased during heat stress but declined after recovery. Activities of glutathione peroxidase (GPX), ascorbate peroxidase (APX), monodehydroascorbate reductase, dehydroascorbate reductase (DHAR) and glutathione reductase (GR) decreased during heat stress but an opposite trend for most of these enzymes was observed after recovery. Heat stress also resulted in significant increases in the activities of glyoxalase enzymes (Gly I and Gly II). In contrast, exogenous Spd protected rice seedlings from heat-induced damage as marked by lower levels of MDA, H₂O₂, and Pro content coupled with increased levels of AsA, GSH, FW, Chl, and AsA and GSH redox status. After recovery, Spd-pretreated heat-exposed seedlings displayed higher activities of SOD, CAT, GPX, GST APX, DHAR and GR as well as of Gly I and Gly II. In addition, polyamine analysis revealed that exogenously applied Spd significantly elevated the levels of free and soluble conjugated Spd. Therefore, we conclude from our results that heat exposure provoked an oxidative burden while enhancement of the antioxidative and glyoxalase systems by Spd rendered rice seedlings more tolerant to heat stress. Further, co-induction of the antioxidative and glyoxalase systems was closely associated with Spd mediated enhanced level of GSH.     

Oxidative stress, protein carbonylation, proteolysis and antioxidative defense system as a model for depicting water deficit tolerance in Indica rice seedlings

Water deficit is an important constraint to rice (Oryza sativa L.) productivity. The present study was undertaken to investigate whether the level of oxidative stress, carbonylation of proteins, proteolysis and status of antioxidative defense could serve as a model to distinguish water deficit tolerant and sensitive rice cultivars. When 10-day-grown seedlings of two rice cultivars, Malviya-36 (drought-sensitive) and Brown Gora (drought-tolerant) were subjected to ?1.0 and ?2.1 MPa water deficit treatments for 24–72 h with polyethylene glycol 6000 in the medium, a greater decline in the growth of the seedlings and levels of leaf water potential, relative water content, Chl a, Chl b, carotenoids and greater increase in leaf water loss were observed in the sensitive cultivar than the tolerant. Under similar level of water deficit seedlings of sensitive cultivar showed higher level of superoxide anion generation, H₂O₂, lipid peroxidation and proteolysis in roots as well as shoots compared to the tolerant. Drought-tolerant cultivar had higher constitutive level of antioxidative enzymes superoxide dismutase and catalase and the activities of these two enzymes alongwith of guaiacol peroxidase showed greater increase in this cultivar under water deficit compared to the sensitive. A significant decline in the level of protein thiol and a higher increase in protein carbonyls content, also confirmed by protein gel blot analysis with an antibody against 2,4-dinitrophenylhydrazine was observed in the seedlings of drought sensitive cv. Malviya-36 compared to the tolerant cv. Brown Gora when subjected to similar level of water deficit. Seedlings of drought sensitive cultivar, under water deficit, showed higher proteolytic activity, higher number of in-gel activity stained proteolytic bands and higher expression of oxidized proteins in roots compared to the tolerant cultivar. Results suggest that poor capacity of antioxidative enzymes could be, at least partly, correlated with water deficit sensitivity of sensitive cultivar and that higher activity of antioxidative enzymes superoxide dismutase, catalase, guaiacol peroxidase, low proteolytic activity, lower level of protein carbonyls and protein thiolation could serve as a model to depict water deficit tolerance in Indica rice seedlings.     

Arsenite treatment induces oxidative stress,upregulates antioxidant system,and causes phytochelatin synthesis in rice seedlings

The effects of arsenite treatment on generation of reactive oxygen species, induction of oxidative stress, response of antioxidative system, and synthesis of phytochelatins were investigated in two indica rice (Oryza sativa L.) cvs. Malviya-36 and Pant-12 grown in sand cultures for a period of 5–20 days. Arsenite (As₂O₃; 25 and 50 μM) treatment resulted in increased formation of superoxide anion (O₂^.−), elevated levels of H₂O₂ and thiobarbituric acid reactive substances, showing enhanced lipid peroxidation. An enhanced level of ascorbate (AA) and glutathione (GSH) was observed irrespective of the variation in the level of dehydroascorbate (DHA) and oxidized glutathione (GSSG) which in turn influenced redox ratios AA/DHA and GSH/GSSG. With progressive arsenite treatment, synthesis of total acid soluble thiols and phytochelatins (PC) increased in the seedlings. Among antioxidative enzymes, the activities of superoxide dismutase (EC 1.15.1.1), catalase (EC 1.11.1.6), total ascorbate peroxidase (APX, EC 1.11.1.11), chloroplastic ascorbate peroxidase, guaiacol peroxidase (EC 1.11.1.7), monodehydroascorbate reductase (EC 1.6.5.4), and glutathione reductase (EC 1.6.4.2) increased in arsenite treated seedlings, while dehyroascorbate reductase (EC 1.8.5.1) activity declined initially during 5–10 days and increased thereafter. Results suggest that arsenite treatment causes oxidative stress in rice seedlings, increases the levels of many enzymatic and non-enzymatic antioxidants, and induces synthesis of thiols and PCs, which may serve as important components in mitigating arsenite-induced oxidative damage.     

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.     

Effect of aspartic acid on physiological characteristics and gene expression of salt exclusion in Tartary buckwheat under salt stress

Salt-tolerant variety Chuanqiao No. 1 and salt-sensitive variety Chuanqiao No. 2 of Tartary buckwheat were used as experimental materials. The effect of aspartic acid on seed germination, physiological characteristics of seedlings and gene expression of salt exclusion in Tartary buckwheat were studied under NaCl stress of 150 mM. The results showed that the aspartic acid treatment could restore the seed germination rate and root vigor of seedlings to the control with non-damage level in salt-tolerant Tartary buckwheat variety under salt stress, and the salt-sensitive variety was increased greatly. Spraying aspartic acid had some protective effects on cell membrane of leaves in Tartary buckwheat under salt stress, and the protective effects were more obviously on salt-sensitive variety, and that could restore the activity of SOD and CAT of leaves to the control level in salt-tolerant Tartary buckwheat variety under salt stress, and the activity of antioxidant enzymes in salt-sensitive variety was increased significantly. The relative expression of FtNHX1 and FtSOS1 genes was increased significantly under salt stress, and that of FtNHX1 gene in salt-tolerant and salt-sensitive varieties was reached the maximum expression level at 12 h and 24 h respectively, while that of FtSOS1 gene in salt-tolerant and salt-sensitive varieties was reached the maximum expression level at 12 h, and the salt-tolerant variety was increased greatly. After spraying aspartic acid, the relative expression of FtNHX1 and FtSOS1 genes was increased more obviously. The relative expression of FtNHX1 gene in salt-tolerant and salt-sensitive varieties was reached the maximum expression level at 12 h, while that of FtSOS1 gene was reached the maximum expression level at 12 h and 24 h respectively, and that in salt-tolerant variety was increased especially more, indicating that spraying aspartic acid on gene expression of salt exclusion in salt-tolerant variety of Tartary buckwheat has a better effect under salt stress.     

Redox metabolic and molecular parameters for screening drought tolerant indigenous aromatic rice cultivars

The present work makes an effort to assess and standardize some redox metabolic and molecular parameters for screening drought tolerant indigenous aromatic rice cultivars of West Bengal, India. PEG-induced dehydration stress during early germination caused disruption of redox-homeostasis and oxidative damage in four IARVs (Jamainadu, Tulaipanji, Sitabhog and Badshabhog) by enhancing the accumulation of pro-oxidants [assessed in terms of oxidation of 2′,7′-dichlorofluorescindiacetate (DCFDA), accumulation of \({\text{O}}_{2}^{ \cdot - }\) and H₂O₂ and in situ staining of reactive oxygen species (ROS) in germinating tissue], significant reduction of antioxidative defence (total antioxidant and radical scavenging capacity, total thiol content and activities of antioxidative defence enzymes) and aggravating protein oxidation and lipid peroxidation (assessed in terms of free carbonyl content and accumulation of thiobarbituric acid reactive substances). When compared between the indigenous aromatic rice cultivars, a clear trend in differential redox regulatory properties in which ROS-antioxidant interaction acts at metabolic interface for redox homeostasis was observed in the order Badshabhog > Tulaipanji > Sitabhog > Jamainadu. Moreover, when the efficacy of ascorbate–glutathione cycle for scavenging H₂O₂ generated during dehydration stress was assessed and compared between the landraces exposed to PEG-induced dehydration stress in germinating tissue, it also exhibited almost the same trend with the landrace Tulaipanji and Badsabhog exhibiting maximum and Jamainadu the minimum efficiencies of the redox cycle. The indigenous aromatic rice cultivars Tulaipanji and Badsabhog resist dehydration stress better than the other two landraces due to its early preparedness to combat oxidative stress by up-regulating expression of genes of some enzymes of ascorbate–glutathione cycle along with some other antioxidative enzymes. A model of redox homeostasis in which ROS-antioxidant (ascorbate–glutathione system) acts at metabolic interface for up-regulation of antioxidative gene expression necessary for differential drought stress tolerance among the indigenous aromatic rice varieties is suggested.     

An inductive pulse of hydrogen peroxide pretreatment restores redox-homeostasis and oxidative membrane damage under extremes of temperature in two rice cultivars

Imbibitional heat and chilling stress caused disruption of redox-homeostasis and oxidative damage to newly assembled membrane system by aggravating membrane lipid peroxidation and protein oxidation [measured in terms of thiobarbituric acid reactive substances (TBARS), free carbonyl content (C=O groups) and membrane protein thiol level (MPTL)] along with concomitant increase in accumulation of reactive oxygen species (superoxide and hydrogen peroxide) and significant reduction of antioxidative defense (assessed in terms of total thiol content and activities of superoxide dismutase, catalase, ascorbate peroxidase and glutathione reductase) in both the salt sensitive (Ratna) and resistant (SR 26B) germinating tissues of rice cultivars. When compared, salt resistant cultivar SR 26B found to suffer significantly less oxidative membrane damage as compared to salt sensitive cultivar Ratna. Treatment with low titer of hydrogen peroxide caused significant reversal in oxidative damages to the newly assembled membrane system imposed by imbibitional heat and chilling stress (evident from the data of TBARS, C=O, MPTL, ROS accumulation, membrane permeability status, membrane injury index and oxidative stress index) in seedlings of both the cultivars of rice (Ratna and SR 26B). Imbibitional H₂O₂ pretreatment also caused up-regulation of antioxidative defense (activities of superoxide dismutase, catalase, ascorbate peroxidase, glutathione reductase and total thiol content) in the heat and chilling stress-raised seedlings of experimental rice cultivars. When the parameters of early growth performances were assessed (in terms of relative growth index, biomass accumulation and vigor index), it clearly exhibited significant improvement of early growth performances of both the rice cultivars. Better response towards H₂O₂-mediated acclamatory performances and restoration of redox- homeostasis under extremes of temperature were noticed in salt sensitive rice cultivar Ratna compared to salt resistant SR 26B. Taken as a whole, the result suggests the significance of the role of ‘inductive pulse’ of H₂O₂ in acclimatizing adverse temperature stress by restoration of redox-homeostasis and mitigation of oxidative membrane protein and lipid damages during the recovery phase of post-germination event.     

Exogenous application of thiamin promotes growth and antioxidative defense system at initial phases of development in salt-stressed plants of two maize cultivars differing in salinity tolerance

The effects of thiamin (Thi) applied as seed soaking or foliar spray on some key physiological parameters were investigated in two differentially salt-responsive maize (Zea mays L.) cultivars, DK 5783 and Apex 836 F1, exposed to saline stress in two different experiments. An initial experiment (germination experiment) was designed to identify appropriate doses of Thi which could lessen the deleterious effects of salt on plants and screen all available maize cultivars for their differential tolerance to salt stress (100 mM NaCl). The seeds of nine maize cultivars were soaked for 24 h in solutions containing six levels of Thi (25, 50, 75, 100, 125 and 150 mg l^?1). Based on the results obtained from the germination experiment, maize cultivar DK 5783 was found to be the most salt tolerant and Apex 836 as the most sensitive cultivar. Also, of six Thi levels used, two levels (100 and 125 mg l^?1) were chosen for subsequent studies. In the second experiment (glasshouse experiment), two maize cultivars, DK 5783 (salt tolerant) and Apex 836 (salt sensitive) were subjected to saline regime (100 mM NaCl) and two levels of Thi (100 and 125 mg l^?1) applied as foliar spray. Salt stress markedly suppressed shoot and root dry mass, total chlorophylls (“a” + “b”), leaf water potential and maximum fluorescence yield (Fv/Fm) in the plants of both maize cultivars, but it increased proline accumulation, leaf osmotic pressure, malondialdehyde (MDA) and hydrogen peroxide (H₂O₂) concentrations, electrolyte leakage (EL) as well as activities of some key antioxidant enzymes, superoxide dismutase (SOD; EC. 1.15.1.1), peroxidase (POD; EC. 1.11.1.7) and catalase (CAT; EC. 1.11.1.6). Salt-induced reduction in plant growth parameters was higher in the salt-sensitive cultivar, Apex 836, which was found to be associated with relatively increased EL, and MDA and H₂O₂ levels, and decreased activities of the key antioxidant enzymes. Application of Thi as seed soaking or foliar spray partly mitigated the deleterious effects of salinity on plants of both maize cultivars. The most promising effect of Thi on alleviation of adverse effects of salt stress on maize plants was found when it was applied as foliar spray at 100 mg l^?1. Thiamin application considerably reduced tissue Na⁺ concentration, but improved those of N, P, Ca²⁺ and K⁺ in the salt-stressed maize plants. Exogenously applied thiamin-induced growth improvement in maize plants was found to be associated with reduced membrane permeability, MDA and H₂O₂ levels, and altered activities of some key antioxidant enzymes such as CAT, SOD and POD as well as increased photosynthetic pigment concentration under saline regime.