Evren Yildiztugay

Elucidation of physiological and biochemical mechanisms of an endemic halophyte Centaurea tuzgoluensis under salt stress.

In this study, physiological and biochemical responses of Centaurea tuzgoluensis, a Turkish endemic halophyte, to salinity were studied. Therefore, the changes in shoot growth, leaf relative water content (RWC), ion concentrations, lipid peroxidation, hydroxyl (OH·) radical scavenging activity, proline (Pro) content, and antioxidant system [superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) and glutathione reductase (GR)] were investigated. The 60 days (d) old C. tuzgoluensis seedlings were subjected to 0, 150 and 300 mM NaCl for 7 d and 14 d. The relative shoot growth was generally did not change in the 150 mM NaCl, but reduced with 300 mM NaCl stress at 7 d and 14 d. RWC was higher in 150 mM NaCl-treated leaves than that of 300 mM NaCl. Salinity decreased K⁺/Na⁺ ratio, but increased Na⁺, Cl⁻, Ca⁺² and Na⁺/Cl⁻ ratio in the leaves. On the other hand, it did not change or increase the K⁺ content at 150 and 300 mM NaCl, respectively. MDA content in the 150 and 300 mM NaCl-treated leaves remained close to control at 7 d. This was related to enhanced activities of SOD, CAT, APX and GR enzymes, and their isoenzymes especially Fe-SOD in the leaves. On the other hand, the higher sensitivity to 300 mM NaCl at 14 d was associated with inadequate increase in antioxidant enzymes and the decreased OH radical scavenging activity. All these results suggest that C. tuzgoluensis has different antioxidant metabolisms between short- (7 d) and long-term (14 d) salt treatments and salinity tolerance of C. tuzgoluensis might be closely related to increased capacity of antioxidative system to scavenge reactive oxygen species (ROS) and accumulation of osmoprotectant proline under salinity conditions.

Changes in the alternative electron sinks and antioxidant defence in chloroplasts of the extreme halophyte Eutrema parvulum (Thellungiella parvula) under salinity

BACKGROUND AND AIMS Eutrema parvulum (synonym, Thellungiella parvula) is an extreme halophyte that thrives in high salt concentrations (100-150 mm) and is closely related to Arabidopsis thaliana. The main aim of this study was to determine how E. parvulum uses reactive oxygen species (ROS) production, antioxidant systems and redox regulation of the electron transport system in chloroplasts to tolerate salinity. METHODS Plants of E. parvulum were grown for 30 d and then treated with either 50, 200 or 300 mm NaCl. Physiological parameters including growth and water relationships were measured. Activities of antioxidant enzymes were determined in whole leaves and chloroplasts. In addition, expressions of chloroplastic redox components such as ferrodoxin thioredoxin reductases (FTR), NADPH thioredoxin reductases (NTRC), thioredoxins (TRXs) and peroxiredoxins (PRXs), as well as genes encoding enzymes of the water-water cycle and proline biosynthesis were measured. KEY RESULTS Salt treatment affected water relationships negatively and the accumulation of proline was increased by salinity. E. parvulum was able to tolerate 300 mm NaCl over long periods, as evidenced by H2O2 content and lipid peroxidation. While Ca(2+) and K(+) concentrations were decreased by salinity, Na(+) and Cl(-) concentrations increased. Efficient induction of activities and expressions of water-water cycle enzymes might prevent accumulation of excess ROS in chloroplasts and therefore protect the photosynthetic machinery in E. parvulum. The redox homeostasis in chloroplasts might be achieved by efficient induction of expressions of redox regulatory enzymes such as FTR, NTRC, TRXs and PRXs under salinity. CONCLUSIONS E. parvulum was able to adapt to osmotic stress by an efficient osmotic adjustment mechanism involving proline and was able to regulate its ion homeostasis. In addition, efficient induction of water-water cycle enzymes and other redox regulatory components such as TRXs and PRXs in chloroplasts were able to protect the chloroplasts from salinity-induced oxidative stress.

The role of antioxidant responses on the tolerance range of extreme halophyte Salsola crassa grown under toxic salt concentrations

Salsola crassa (Amaranthaceae) is an annual halophytic species and naturally grows in arid soils that are toxic to most plants. In order to study the effects of salinity on their antioxidant system and to determine the tolerance range against salt stress, S. crassa seeds were grown with different concentrations of NaCl (0, 250, 500, 750, 1000, 1250 and 1500mM) for short (15d) and long-term (30d). Results showed that growth (RGR), water content (RWC) and osmotic potential (ΨΠ) decreased and, proline content (Pro) increased at prolonged salt treatment. Unlike K(+) and Ca(2+) contents, S. crassa highly accumulated Na(+) and Cl(-) contents. Chlorophyll fluorescence (Fv/Fm) only decreased in response to 1500mM NaCl at 30d. No salt stimulation of superoxide anion radical (O2(•-)) content was observed in plants treated with the range of 0-500mM NaCl during the experimental period. NaCl increased superoxide dismutase (SOD) activity depending on intensities of Mn-SOD and Fe-SOD isozymes except in 1500mM NaCl-treated plants at 30d. In contrast to catalase (CAT), peroxidase (POX) activity increased throughout the experiment. Also, salinity caused an increase in glutathione reductase (GR) and glutathione peroxidase (GPX) and decreased in ascorbate peroxidase (APX), monodehydroascorbate reductase (MDHAR) and dehydroascorbate reductase (DHAR) at 15d. Both total ascorbate (tAsA) and glutathione (tGlut) contents significantly increased in treated plants with 1000-1500mM NaCl at 15d. After 0-1000mM NaCl stress, H2O2 and TBARS contents were similar to control groups at 15d, which were consistent with the increased antioxidant activity (POX, GR and GPX). However, H2O2 content was more pronounced at 30d. Therefore, S. crassa exhibited inductions in lipid peroxidation (TBARS content) in response to extreme salt concentrations. These results suggest that S. crassa is tolerant to salt-induced damage at short-term treatments as well as extreme salt concentrations.

Halophytes as a source of salt tolerance genes and mechanisms: a case study for the Salt Lake area, Turkey

The worst case scenario of global climate change predicts both drought and salinity would be the first environmental factors restricting agriculture and natural ecosystems, causing decreased crop yields and plant growth that would directly affect human population in the next decades. Therefore, it is vital to understand the biology of plants that are already adapted to these extreme conditions. In this sense, extremophiles such as the halophytes offer valuable genetic information for understanding plant salinity tolerance and to improve the stress tolerance of crop plants. Turkey has ecological importance for its rich biodiversity with up to 3700 endemic plants. Salt Lake (Lake Tuz) in Central Anatolia, one of the largest hypersaline lakes in the world, is surrounded by salty marshes, with one of the most diverse floras in Turkey, where arid and semiarid areas have increased due to low rainfall and high evaporation during the summer season. Consequently, the Salt Lake region has a large number of halophytic, xerophytic and xero-halophytic plants. One good example is Eutrema parvulum (Schrenk) Al-Shehbaz & Warwick, which originates from the Salt Lake region, can tolerate up to 600mM NaCl. In recent years, the full genome of E. parvulum was published and it has been accepted as a model halophyte due to its close relationship (sequence identity in range of 90%) with Arabidopsis thaliana (L. Heynh.). In this context, this review will focus on tolerance mechanisms involving hormone signalling, accumulation of compatible solutes, ion transporters, antioxidant defence systems, reactive oxygen species (ROS) signalling mechanism of some lesser-known extremophiles growing in the Salt Lake region. In addition, current progress on studies conducted with E. parvulum will be evaluated to shed a light on future prospects for improved crop tolerance.

The impact of selenium application on enzymatic and non-enzymatic antioxidant systems in Zea mays roots treated with combined osmotic and heat stress

ABSTRACT Selenium (Se), regarded as an antioxidant, has been found beneficial for plants growing under stressed conditions. To investigate whether the Se application helps to improve stress tolerance, sodium selenite (Na2SeO3 · 5H2O, 5–15 μM) was hydroponically applied to Zea mays variety OSSK-713-roots under heat and/or PEG-induced osmotic stress (25% PEG-6000) for 8 h. The individual/combined stress caused accumulation of reactive oxygen species (ROS). While only superoxide dismutase (SOD) increased with heat stress alone, the activities of SOD, catalase (CAT) and ascorbate peroxidase (APX) increased under PEG exposure. The combination of these stresses resulted in an induction of both SOD and CAT activities. Lipid peroxidation (TBARS) levels were also high in all the stress treatments, especially under the combination treatment. Addition of Se not only improved the activities of SOD, APX and glutathione reductase (GR) in stress-treated roots, but it also changed the activities of monodehydroascorbate reductase (MDHAR) and dehydroascorbate reductase (DHAR). The findings reveal that Se has a positive effect on heat and/or osmotic stress mitigation mainly by regulating the ascorbate-glutathione cycle, especially in PEG-treated plants. Under the combined stress treatment, addition of 5 µM of exogenous Se was most effective.

Profiling of rutin-mediated alleviation of cadmium-induced oxidative stress in Zygophyllum fabago.

Zygophyllum fabago grows in arid, saline soil, or disturbed sites, such as former industrial or mining areas. This species is able to grow in coarse mineral substrates contaminated with heavy metals. To investigate the effects of the flavonoid rutin (Rtn) on certain heavy metal stress responses such as antioxidant defense systems and water status, seedlings were subjected to 100 and 200 μM CdCl2 treatment without or with 0.25 and 1 mM Rtn for 7 and 14 d (days). Cd stress decreased growth (RGR), water content (RWC), leaf osmotic potential (ΨΠ), and chlorophyll fluorescence, all of which could be partly alleviated by addition of Rtn. Activities of superoxide dismutase, peroxidase (POX), ascorbate peroxidase, and glutathione reductase increased within the first 7 d after exposure to Cd. However, failure of antioxidant defense in the scavenging of reactive oxygen species (ROS) was evidenced by an abnormal rise in superoxide anion radical ( O2•− ) and hydrogen peroxide contents and a decline in hydroxyl radical (OH•) scavenging activity, resulting in enhancement of lipid peroxidation (TBARS) as a marker of Cd‐induced oxidative stress. However, exogenously applied Rtn considerably improved the stress tolerance of plants via a reduction in Cd accumulation, modulation of POX activity, increase of proline (Pro) content, decrease in TBARS and ROS content and consequent lowering of oxidative damage of membrane. Overall, 0.25 and 1 mM Rtn could protect Z. fabago from the harmful effects of 100 μM Cd‐induced oxidative stress throughout the experiment.

The humic acid-induced changes in the water status, chlorophyll fluorescence and antioxidant defense systems of wheat leaves with cadmium stress

The using of bio-stimulant in plants grown under stress conditions for enhancing nutrition efficiency and crop quality traits is an effective approach. One of the bio-stimulants, humus material, is defined as humic acid (HA). HA application as a promotion of plant growth to plants grown in the heavy metals-contaminated soils has promised hope in terms of effects on plants but the its limiting effect is the application dose. Therefore, the wheat seedlings were grown in hydroponic culture for 21 d and the various concentrations of humic acid (HA; 750 or 1500 mg L-1) were treated alone or in combination with cadmium (Cd) stress (100 or 200 μM) for 7 d. The results showed that after Cd stress treatment, water content (RWC), osmotic potential (ΨΠ) and chlorophyll fluorescence parameters decreased and proline content (Pro) increased for 7 d. In spite of activated peroxidase (POX) and ascorbate peroxidase (APX), stress induced the toxic levels of hydrogen peroxide (H2O2) accumulation. Cd stress triggered lipid peroxidation (TBARS content). HA application successfully eliminated the negative effects of stress on RWC, ΨΠ and photosynthetic parameters. In the presence of HA under stress, the increased activation of superoxide dismutase (SOD), catalase (CAT) and NADPH-oxidase (NOX) enzymes and ascorbate, glutathione and GSH/GSSG ratio observed. Only 750 mg L-1 HA under stress conditions induced the activities of monodehydroascorbate reductase (MDHAR) and dehydroascorbate reductase (DHAR), and dehydroascorbate (DHA) content. After the combined application of HA and Cd stress, the low contents of H2O2 and TBARS maintained in wheat leaves. Hence, HA successfully eliminated the toxicity of Cd stress by modulating the water status, photosynthetic apparatus and antioxidant activity in wheat leaves.

Improvement of cold stress resistance via free radical scavenging ability and promoted water status and photosynthetic capacity of gallic acid in soybean leaves

Plant polyphenols exhibit a wide variety of biological activities such as antimutagenicity, anticarcinogenicity and antioxidative activity. There is no report whether gallic acid (GLA), a naturally occurring plant phenol, is able to activate the plant defense system under cold stress. For this purpose, after soybean (Glycine max) was hydroponically grown for 3 weeks, seedlings were treated with gallic acid (GLA; 1 and 2 mM) and cold stress (5 oC and 10 oC) and GLA and stress combination for 72 h. The inhibition in growth, water content (RWC), osmotic potential (Ψп) and photosynthetic activity observed under stress and was more at the lowest temperature. Stress also elicited the accumulation of proline (Pro) only at 5 oC. While the capacity to maintain high growth, RWC, Ψп and photosynthetic efficiency was observed in GLA-treated plants under stress, Pro accumulation could not achieve with GLA plus stress. Any increase in total activities of superoxide dismutase (SOD) and catalase (CAT) induced by stress treatments determined. The lower cold stress caused an increase in the activities of ascorbate peroxidase (APX), glutathione reductase (GR) and NADPH oxidase (NOX). GLA treatment under stress (especially at 5 oC) could supply the increased activities of SOD, CAT, APX and GR. Also, exogenous GLA application to stress-treated plants increased the enzyme activities in ascorbate-glutathione cycle such as monodehydroascorbate reductase (MDHAR) and dehydroascorbate reductase (DHAR) and, contents of ascorbate (AsA) and glutathione. After GLA application under stress, it is observed reduction in hydrogen peroxide (H2O2) and the levels of lipid peroxidation (TBARS), and induction of hydroxyl radical (OH ●) scavenging. Our results suggest that GLA is a potent inducer for induction of the scavenging activity of radicals as well as effectively usage of water status and photosynthetic capacity.