Baris Uzilday

Comparison of ROS formation and antioxidant enzymes in Cleome gynandra (C4) and Cleome spinosa (C3) under drought stress

Differences between antioxidant responses to drought in C(3) and C(4) plants are rather scanty. Even, we are not aware of any research on comparative ROS formation and antioxidant enzymes in C(3) and C(4) species differing in carboxylation pathway of same genus which would be useful to prevent other differences in plant metabolism. With this aim, relative shoot growth rate, relative water content and osmotic potential, hydrogen peroxide (H(2)O(2)) content and NADPH oxidase (NOX) activity, antioxidant defence system (superoxide dismutase (SOD), catalase (CAT), peroxidase (POX), ascorbate peroxidase (APX), glutathione reductase (GR) enzymes and their isoenzymes), CAT1 mRNA level, and lipid peroxidation in seedlings of Cleome spinosa (C(3)) and Cleome gynandra (C(4)) species of Cleome genus exposed to drought stress for 5 and 10 day (d) were comparatively investigated. Constitutive levels of antioxidant enzymes (except SOD) were consistently higher in C. spinosa than in C. gynandra under control conditions. CAT1 gene expression in C. spinosa was correlated with CAT activity but CAT1 gene expression in C. gynandra at 10 d did not show this correlation. Drought stress caused an increase in POX, CAT, APX and GR in both species. However, SOD activity was slightly decreased in C. gynandra while it was remained unchanged or increased on 5 and 10 d of stress in C. spinosa, respectively. Parallel to results of malon dialdehyde (MDA), H(2)O(2) content was also remarkably increased in C. spinosa as compared to C. gynandra under drought stress. These results suggest that in C. spinosa, antioxidant defence system was insufficient to suppress the increasing ROS production under stress condition. On the other hand, in C. gynandra, although its induction was lower as compared to C. spinosa, antioxidant system was able to cope with ROS formation under drought stress.

Endoplasmic reticulum stress triggers ROS signalling, changes the redox state, and regulates the antioxidant defence of Arabidopsis thaliana

Summary Endoplasmic reticulum stress, which is induced by tunicamycin, triggers reactive oxygen species signalling via NADPH oxidase activity and also regulates the antioxidant defence system in Arabidopsis thaliana.

Strategies of ROS regulation and antioxidant defense during transition from C3 to C4 photosynthesis in the genus Flaveria under PEG-induced osmotic stress

In the present study, we aimed to elucidate how strategies of reactive oxygen species (ROS) regulation and the antioxidant defense system changed during transition from C₃ to C₄ photosynthesis, by using the model genus Flaveria, which contains species belonging to different steps in C₄ evolution. For this reason, four Flaveria species that have different carboxylation mechanisms, Flaveria robusta (C₃), Flaveria anomala (C₃-C₄), Flaveria brownii (C₄-like) and Flaveria bidentis (C₄), were used. Physiological (growth, relative water content (RWC), osmotic potential), and photosynthetical parameters (stomatal conductance (g(s)), assimilation rate (A), electron transport rate (ETR)), antioxidant defense enzymes (superoxide dismutase (SOD), catalase (CAT), peroxidase (POX), ascorbate peroxidase (APX), glutathione reductases(GR)) and their isoenzymes, non-enzymatic antioxidant contents (ascorbate, glutathione), NADPH oxidase (NOX) activity, hydrogen peroxide (H₂O₂) content and lipid peroxidation levels (TBARS) were measured comparatively under polyethylene glycol (PEG 6000) induced osmotic stress. Under non-stressed conditions, there was a correlation only between CAT (decreasing), APX and GR (both increasing) and the type of carboxylation pathways through C₃ to C₄ in Flaveria species. However, they responded differently to PEG-induced osmotic stress in regards to antioxidant defense. The greatest increase in H₂O₂ and TBARS content was observed in C₃ F. robusta, while the least substantial increase was detected in C₄-like F. brownii and C₄ F. bidentis, suggesting that oxidative stress is more effectively countered in C₄-like and C₄ species. This was achieved by a better induced enzymatic defense in F. bidentis (increased SOD, CAT, POX, and APX activity) and non-enzymatic antioxidants in F. brownii. As a response to PEG-induced oxidative stress, changes in activities of isoenzymes and also isoenzymatic patterns were observed in all Flaveria species, which might be related to ROS produced in different compartments of cells.

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.

Interplay between the unfolded protein response and reactive oxygen species: a dynamic duo

Secretory proteins undergo modifications such as glycosylation and disulphide bond formation before proper folding, and move to their final destination via the endomembrane system. Accumulation of unfolded proteins in the endoplasmic reticulum (ER) due to suboptimal environmental conditions triggers a response called the unfolded protein response (UPR), which induces a set of genes that elevate protein folding capacity in the ER. This review aims to establish a connection among ER stress, UPR, and reactive oxygen species (ROS), which remains an unexplored topic in plants. For this, we focused on mechanisms of ROS production originating from ER stress, the interaction between ER stress and overall ROS signalling process in the cell, and the interaction of ER stress with other organellar ROS signalling pathways such as of the mitochondria and chloroplasts. The roles of the UPR during plant hormone signalling and abiotic and biotic stress responses are also discussed in connection with redox and ROS signalling.

Changes in redox regulation during transition from C 3 to single cell C 4 photosynthesis in Bienertia sinuspersici

Bienertia sinuspersici performs single cell C4 photosynthesis without Kranz anatomy. Peripheral and central cytoplasmic compartments in a single chlorenchyma cell act as mesophyll cells and bundle sheath cells. Development of this specialized mechanism is gradual during plant development. Young leaves perform C3 photosynthesis, while mature leaves have complete C4 cycle. The aim of this work was to investigate changes in redox regulation and antioxidant defence during transition from C3 to single cell C4 photosynthesis in B. sinuspersici leaves. First, we confirmed gradual development of C4 with protein blot and qRT-PCR analysis of C4 enzymes. After this activities and isoenzymes of superoxide dismutase (SOD), catalase (CAT), peroxidase (POX), ascorbate peroxidase (APX), glutathione reductase (GR), dehydroascorbate reductase (DHAR) and H2O2 and TBARS and glutathione pool and redox status (GSH/GSSG) were determined in young, developing and mature leaves during transition from C3 to single cell C4 photosynthesis. Activities of SOD, APX and POX decrease, while GR and DHAR were increased. However, most striking results were the changes in isoenzyme patterns of SOD, CAT and GR which were gradual through transition to C4 photosynthesis.

Reactive oxygen species and redox regulation in mesophyll and bundle sheath cells of C4 plants

Redox regulation, antioxidant defence, and reactive oxygen species (ROS) signalling are critical in performing and tuning metabolic activities. However, our concepts have mostly been developed for C3 plants since Arabidopsis thaliana has been the major model for research. Efforts to convert C3 plants to C4 to increase yield (such as IRRIs C4 Rice Project) entail a better understanding of these processes in C4 plants. Various photosynthetic enzymes that take part in light reactions and carbon reactions are regulated via redox components, such as thioredoxins as redox transmitters and peroxiredoxins. Hence, understanding redox regulation in the mesophyll and bundle sheath chloroplasts of C4 plants is of paramount importance: it appears impossible to utilize efficient C4 photosynthesis without understanding its exact redox needs and the regulation mechanisms used during light reactions. In this review, we discuss current knowledge on redox regulation in C3 and C4 plants, with special emphasis on the mesophyll and bundle sheath differences that are found in C4. In these two cell types in C4 plants, linear and cyclic electron transport in the chloroplasts operate differentially when compared to C3 chloroplasts, changing the redox needs of the cell. Therefore, our focus is on photosynthetic light reactions, ROS production dynamics, antioxidant defence, and thiol-based redox regulation, with the aim of providing an overview of our current knowledge.

Endoplasmic reticulum stress regulates glutathione metabolism and activities of glutathione related enzymes in Arabidopsis

Stress conditions generate an extra load on protein folding machinery in the endoplasmic reticulum (ER) and if the ER cannot overcome this load, unfolded proteins accumulate in the ER lumen, causing ER stress. ER lumen localised protein disulfide isomerase (PDI) catalyses the generation of disulfide bonds in conjugation with ER oxidoreductase1 (ERO1) during protein folding. Mismatched disulfide bonds are reduced by the conversion of GSH to GSSG. Under prolonged ER stress, GSH pool is oxidised and H2O2 is produced via increased activity of PDI-ERO1. However, it is not known how glutathione metabolism is regulated under ER stress in plants. So, in this study, ER stress was induced with tunicamycin (0.15, 0.3, 0.45μg mL-1 Tm) in Arabidopsis thaliana (L.) Heynh. Glutathione content was increased by ER stress, which was accompanied by induction of glutathione biosynthesis genes (GSH1, GSH2). Also, the apoplastic glutathione degradation pathway (GGT1) was induced. Further, the activities of glutathione reductase (GR), dehydroascorbate reductase (DHAR), glutathione peroxidase (GPX) and glutathione S-transferase (GST) were increased under ER stress. Results also showed that chloroplastic GPX genes were specifically downregulated with ER stress. This is the first report on regulation of glutathione metabolism and glutathione related enzymes in response to ER stress in plants.

Redox Regulation and Antioxidant Defence During Abiotic Stress: What Have We Learned from Arabidopsis and Its Relatives?

Abiotic stress conditions are a global constraint that affects plant growth and crop yield worldwide, and this phenomenon is expected to be increased in the forthcoming future due to global climate change. Arabidopsis thaliana is the model organism for plant science since the early 1990s, and its genome has been known for more than a decade. Studies conducted with Arabidopsis created a foundation that could be transferred and used in its close relatives to similarity of genetic sequences. Up to now, studies on A. thaliana gave deep insight into different abiotic stress tolerance mechanisms. However, A. thaliana is not a stress-tolerant plant species. Therefore some of the stress tolerance mechanisms that are used by its stress-tolerant relatives might not even be observed in Arabidopsis. This chapter focused on reactive oxygen species (ROS) production during environmental stress and antioxidant defence systems activated against it in A. thaliana and its close relatives such as Thellungiella sp., A. halleri, Thlaspi sp., Lepidium sativum and Arabis paniculata.