Phytochrome‐mediated regulation of plant respiration and photorespiration

The expression of genes encoding various enzymes participating in photosynthetic and respiratory metabolism is regulated by light via the phytochrome system. While many photosynthetic, photorespiratory and some respiratory enzymes, such as the rotenone‐insensitive NADH and NADPH dehydrogenases and the alternative oxidase, are stimulated by light, succinate dehydrogenase, subunits of the pyruvate dehydrogenase complex, cytochrome oxidase and fumarase are inhibited via the phytochrome mechanism. The effect of light, therefore, imposes limitations on the tricarboxylic acid cycle and on the mitochondrial electron transport coupled to ATP synthesis, while the non‐coupled pathways become activated. Phytochrome‐mediated regulation of gene expression also creates characteristic distribution patterns of photosynthetic, photorespiratory and respiratory enzymes across the leaf generating different populations of mitochondria, either enriched by glycine decarboxylase (in the upper part) or by succinate dehydrogenase (in the bottom part of the leaf).     

Connecting salt stress signalling pathways with salinity‐induced changes in mitochondrial metabolic processes in C3 plants

Salinity exerts a severe detrimental effect on crop yields globally. Growth of plants in saline soils results in physiological stress, which disrupts the essential biochemical processes of respiration, photosynthesis, and transpiration. Understanding the molecular responses of plants exposed to salinity stress can inform future strategies to reduce agricultural losses due to salinity; however, it is imperative that signalling and functional response processes are connected to tailor these strategies. Previous research has revealed the important role that plant mitochondria play in the salinity response of plants. Review of this literature shows that 2 biochemical processes required for respiratory function are affected under salinity stress: the tricarboxylic acid cycle and the transport of metabolites across the inner mitochondrial membrane. However, the mechanisms by which components of these processes are affected or react to salinity stress are still far from understood. Here, we examine recent findings on the signal transduction pathways that lead to adaptive responses of plants to salinity and discuss how they can be involved in and be affected by modulation of the machinery of energy metabolism with attention to the role of the tricarboxylic acid cycle enzymes and mitochondrial membrane transporters in this process.     

Control of plant mitochondrial respiration

Plant mitochondria are characterised by the presence of both phosphorylating (cytochrome) and non-phosphorylating (alternative) respiratory pathways, the relative activities of which directly affect the efficiency of mitochondrial energy conservation. Different approaches to study the regulation of the partitioning of reducing equivalents between these routes are critically reviewed. Furthermore, an updated view is provided regarding the understanding of plant mitochondrial respiration in terms of metabolic control. We emphasise the extent to which kinetic modelling and 'top-down' metabolic control analysis improve the insight in phenomena related to plant mitochondrial respiration. This is illustrated with an example regarding the affinity of the plant alternative oxidase for oxygen.     

Functional expression of oxidative phosphorylation proteins in the rod outer segment disc

The rod Outer Segment (OS) disc, an organelle devoid of mitochondria, is specialized in phototransduction, a process requiring a continual chemical energy supply. We have shown that OS discs express functional mitochondrial electron transport chains, F_oF₁‐ATP synthase and the tricarboxylic acid cycle enzymes, all mitochondrial features. Here, we focus on oxygen consumption and adenosine triphosphate (ATP) synthesis by OS discs analysing electron transport chain I‐III‐IV and II‐II‐IV pathways, supported by reduced nicotinamide adenine dinucleotide and succinate, respectively. Interestingly, respiratory capacity of discs was measurable also in the presence of 3‐hydroxy‐butyrrate, a typical metabolic substrate for the brain. Data were supported by a two‐dimensional electrophoresis analyses conducted as our previous one, but focused to those mitochondrial proteins that are involved in oxidative phosphorylation. Carbonic anhydrase was also found active in OS discs. Moreover, colocalization of Rhodopsin with respiratory complex I and ATP synthase seems a further step in the characterization of some proteins typical of the mitochondrial inner membranes that are expressed in the rod discs. The existence of oxygen utilization in the outer retina, likely supplying ATP for phototransduction, may shed light on some retinal pathologies related to oxidative stress of the outer retina. Copyright © 2013 John Wiley & Sons, Ltd.     

The mechanism of stimulation of respiration by fatty acids in isolated hepatocytes

Addition of fatty acids to isolated hepatocytes raised respiration rate by 92% and raised mitochondrial membrane potential (delta psi m) in situ from 155 to 162 mV suggesting that the increased fuel supply had a greater effect on respiration rate than any increases in processes that consumed mitochondrial protonmotive force (delta p). The relationship between delta psi m and respiration rate was changed by addition of fatty acids or lactate, showing that there was also stimulation of delta p-consuming reactions. In the presence of oligomycin the relationship between delta psi m and respiration rate was unaffected by substrate addition, showing that the kinetics of delta p consumption by the H+ leak across the mitochondrial inner membrane were unchanged. The stimulation of delta p consumers by fatty acids therefore must be in the pathways of ATP synthesis and turnover. Inhibition of several candidate ATP-consuming reactions had little effect on basal or fatty acid-stimulated respiration, and the nature of the ATP turnover reactions in hepatocytes remains speculative. We conclude that fatty acids (and other substrates) stimulate respiration in hepatocytes in two distinct ways. They provide substrate for the electron transport chain, raising delta p and increasing the non-ohmic proton leak across the mitochondrial inner membrane and the rate of oxygen consumption. They also directly stimulate an unidentified delta p-consuming reaction in the cytoplasm. They do not work by uncoupling or by stimulation of intramitochondrial ATP-turnover reactions.     

Lupine embryo axes under salinity stress. II. Mitochondrial proteome response

Germination is the first step of plant growth in plant life cycle. An embryonic radicle protruding the seed coat is the first part of plant which has direct contact with external environment including salt-affected soil. In embryo axes, mitochondria are the main energy producer. To understand better salinity impact on mitochondria functioning, this study was focused on the effect of NaCl stress onto mitochondria proteome. Mitochondria were isolated from yellow lupine (Lupine luteus L. ‘Mister’) embryo axes cultured in vitro for 12 h with 250 and 500 mM NaCl. Two-dimensional gel electrophoresis of mitochondrial proteins isolated from NaCl-treated axes demonstrated significant changes in proteins abundances as a response to salinity treatment. Twenty-one spots showing significant changes in protein expression profiles both under 250 and 500 mM NaCl treatment were selected for tandem mass spectrometry identification. This approach revealed proteins associated with different metabolic processes that represent enzymes of tricarboxylic acid cycle, mitochondrial electron transport chain, enzymes and proteins involved in mitochondria biogenesis and stresses response. Among proteins involved in mitochondria biogenesis, mitochondrial import inner membrane translocase, subunit Tim17/22, mitochondrial-processing peptidase subunit alpha-1, mitochondrial elongation factor Tu and chaperonins CPN60 were revealed. Finally, formate dehydrogenase 1 was found to accumulate in lupine embryo axes mitochondria under salinity. The functions of identified proteins are discussed in relation to salinity stress response, including salinity-induced PCD.     

Stimulation of the electron transport chain in mitochondria isolated from rats treated with mannoheptulose or glucagon

We investigated the kinetics of the mitochondrial respiratory chain, proton leak, and phosphorylating subsystems of liver mitochondria from mannoheptulose-treated and control rats. Mannoheptulose treatment raises glucagon and lowers insulin; it had no effect on the kinetics of the mitochondrial proton leak or phosphorylating subsystems, but the respiratory chain from succinate to oxygen was stimulated. Previous attempts to detect any stimulation of cytochrome c oxidase by glucagon are shown by flux control analysis to have used inappropriate assay conditions. To investigate the site of stimulation of the respiratory chain we measured the relationship between the thermodynamic driving force and respiration rate for the span succinate to coenzyme Q, the cytochrome bc1 complex and cytochrome c oxidase. Hormone treatment of rats altered the kinetics of electron transport from succinate to coenzyme Q in subsequently isolated mitochondria and activated succinate dehydrogenase. The kinetics of electron transport through the cytochrome bc1 complex were not affected. Effects on cytochrome c oxidase were small or nonexistent.     

Creatine kinase of heart mitochondria. Control of oxidative phosphorylation by the extramitochondrial concentrations of creatine and phosphocreatine

Defining how extramitochondrial high-energy phosphate acceptors influence the rates of heart oxidative phosphorylation is essential for understanding the control of myocardial respiration. When the production of phosphocreatine is coupled to electron transport via mitochondrial creatine kinase, the net reaction can be expressed by the balanced equation: creatine + Pi----phosphocreatine + H2O. This suggests that rates of oxygen consumption could be regulated by changes in [creatine], [Pi], or [phosphocreatine], alone or in combination. The effects of altering these metabolites upon mitochondrial rates of respiration were examined in vitro. Rat heart mitochondria were incubated in succinate-containing oxygraph medium (pH 7.2, 37 degrees C) supplemented with five combinations of creatine (1.0-20 mM), phosphocreatine (0-25 mM), and Pi (0.25-5.0 mM). In all cases, the mitochondrial creatine kinase reaction was initiated by additions of 0.5 mM ATP. To emphasize the duality of control, the results are presented as three-dimensional stereoscopic projections. Under physiological conditions, with 5.0 mM creatine, increases in Pi or decreases in phosphocreatine had little influence upon mitochondrial respiration. When phosphocreatine was held constant (15 mM), changes in [creatine] modestly stimulated respiratory rates, whereas Pi again showed little effect. With 1.0 mM Pi, respiration clearly became dependent upon changes in [creatine] and [phosphocreatine]. Initially, respiratory rates increased as a function of [creatine]. However, at [phosphocreatine] values below 10 mM, product "deinhibition" was observed, and respiratory rates rapidly increased to 80% State 3. With 2.0 mM Pi or higher, respiration could be regulated from State 4 to 100% State 3. Overall, the data show how increasing [creatine] and decreasing [phosphocreatine] influence the rates of oxidative phosphorylation when mediated by mitochondrial creatine kinase. Thus, these changes may become secondary cytoplasmic signals regulating heart oxygen consumption.     

Similar nature of inhibition of mitochondrial respiration of heart tissue and malignant cells by methylglyoxal. A vital clue to understand the biochemical basis of malignancy

The effect of methylglyoxal on the oxygen consumption of mitochondria of heart and of several other organs of normal animals of different species has been tested. The results indicate that methylglyoxal (3.5 mM) strongly inhibits ADP-stimulated -oxoglutarate and malate plus pyruvate-dependent respiration of exclusively heart mitochondria of normal animals of different species. Whereas, with the same substrates, but at a higher concentration of methylglyoxal (7.5 mM), the respiration of mitochondria of other organs of normal animals is not inhibited. Methylglyoxal also inhibits the respiration of slices of rat and toad hearts. But this inhibition is less pronounced. However, methylglyoxal (15 mM) fails to have any effect on perfused toad heart. Using rat heart mitochondria as a model, the effect of methylglyoxal on the oxygen consumption was also tested with different respiratory substrates, electron donors at different segments of the mitochondrial respiratory chain and site-spe inhibitors to identify the specific respiratory complex which might be involved in the inhibitory effect of methylglyoxal. The results strongly suggest that methylglyoxal inhibits the electron flow through complex I of rat heart mitochondrial respiratory chain. Moreover, lactaldehyde (0.6 mM), a catabolite of methylglyoxal, can exert a protective effect on the inhibition of rat heart mitochondrial respiration by methylglyoxal (2.5 mM). The effect of methylglyoxal on heart mitochondria as described in the present paper is strikingly similar to the results of our previous work with mitochondria of Ehrlich ascites carcinoma cells and leukemic leukocytes. We have recently proposed a new hypothesis on cancer which suggests that excessive ATP formation in cells may lead to malignancy. The above mentioned similarity apparently provides a solid experimental foundation for the proposed hypothesis which has been discussed.     

植物线粒体呼吸状态的研究方法及其在植物生物学中的应用

线粒体呼吸状态是指当线粒体中呼吸底物及ADP以不同浓度存在时线粒体的呼吸速率,它是用来研究线粒体功能的重要指标。本文结合我们的研究经验,详细阐述了线粒体呼吸状态的研究方法,介绍了该方法在分析线粒体电子传递链功能和氧化磷酸化活性中的应用,此外概述了线粒体呼吸状态的测定和分析方法在植物生理生态研究中的具体应用。     

Ethidium bromide inhibits mitochondrial phosphorylating oxidation.

Ethidium bromide, in addition to combination with mitochondrial nucleic acids, is a phosphorylation inhibitor during glutamate and succinate respiration by mitochondria. Exhaustive washing of ethidium bromide-treated mitochondria did not relieve the inhibition nor significantly decrease the amount of bound dye. Dialysis against a cation exchange resin at 3 degrees for 17 hr removed about 97% of bound dye. This restored phosphorylating capacity to that of untreated mitochondria which had also been dialyzed against the resin. Since state 3 respiration was diminished and state 4 was unaffected by the presence of the acridine dye, and since neither swelling of mitochondria nor release of latent ATPase was observed, then ethidium bromide was not an electron transport inhibitor nor an uncoupler of oxidative phosphorylation. Inhibition of metabolic processes by ethidium bromide may be due in part to depressed generation of mitochondrial ATP.     

Respiration of wheat root cells under simultaneous inhibition of parts I and III of the electron transport chain of mitochondria by rotenone and antimycine A

Respiration of excised roots of 5 day old wheat seedlings with blocked mitochondrial oxidation under simultaneous action of rotenone and antimycine A was studied. A reduced rate of oxygen uptake was observed within the first hour of root treatment inhibitors. However, after a 5 h exposure there was an increase in oxygen uptake, which was prevented by KCN but amplified by malate and ascorbate. The application of inhibitors caused a considerable increase in the respiratory coefficient (RC) up to 2.1, that suggests a significant CO2 release, when the initial sites of mitochondrial electron transport chain were inhibited. RC did not raise, when ascorbate was added in the presence of inhibitors. We assume that inhibition of mitochondrial oxidation at I and III sites of electron transport chain facilitates switching on the alternative paths of reductant translocation to oxygen. Participation of ATPases and redox system of plasma membrane in the response reactions of respiration directed to the restoration of ion, particularly, proton homeostasis in conditions of inhibited mitochondrial oxidation is discussed.     

Responses of the Mitochondrial Respiratory System to Low Temperature in Plants

Low-temperature (LT) stress induces significant changes to plant cells including perturbations of various physio-biochemical and metabolic processes, which impact primary metabolism, respiratory rate, and the ATP production for biosynthesis and growth. Mitochondria from LT-tolerant species respond to LT through remodeling their composition that changes the structural and functional properties of the organelles. In this review, we discuss physiological aspects of mitochondrial respiration rate that are affected by LT, as well as, changes in the abundance of respiratory components under LT. The latter includes components of the phosphorylating and non-phosphorylating pathways and adjustments of mitochondrial membrane composition. Our objective is to provide a detailed overview of the often-contrasting reports of mitochondrial-specific changes and responses to LT and look for consensus themes to explain changes and draw more generally applicable observations about the LT response of plant respiration.     

Sites of inhibition of mitochondrial electron transport in macrophage- injured neoplastic cells

Previous work has shown that injury of neoplastic cells by cytotoxic macrophages (CM) in cell culture is accompanied by inhibition of mitochondrial respiration. We have investigated the nature of this inhibition by studying mitochondrial respiration in CM-injured leukemia L1210 cells permeabilized with digitonin. CM-induced injury affects the mitochondrial respiratory chain proper. Complex I (NADH-coenzyme Q reductase) and complex II (succinate-coenzyme Q reductase) are markedly inhibited. In addition a minor inhibition of cytochrome oxidase was found. Electron transport from alpha-glycerophosphate through the respiratory chain to oxygen is unaffected and permeabilized CM-injured L1210 cells oxidizing this substrate exhibit acceptor control. However, glycerophosphate shuttle activity was found not to occur within CM- injured or uninjured L1210 cells in culture hence, alpha- glycerophosphate is apparently unavailable for mitochondrial oxidation in the intact cell. It is concluded that the failure of respiration of intact neoplastic cells injured by CM is caused by the nearly complete inhibition of complexes I and II of the mitochondrial electron transport chain. The time courses of CM-induced electron transport inhibition and arrest of L1210 cell division are examined and the possible relationship between these phenomena is discussed.     

Differential Responses of Two Wheat Varieties Differing in Salt Tolerance to the Combined Stress of Mn and Salinity

The effect of Mn and NaCl on growth, mineral nutrients and antioxidative enzymes in two tetroploid wheat genotypes differing in salt tolerance was investigated in this study. 100 mM NaCl and Mn stress significantly inhibited plant growth, photosynthesis and Ca uptake, while stimulated ROS accumulation, MDA and proline content in wheat plants, Mn stress also increased SOD, APX, GR and DHAR activities. Durum wheat (AS780) was less affected by 100 mM NaCl and Mn stress than emmer wheat (AS847) due to more proline production, higher antioxidative enzymes activities and less-affected mineral nutrients. Application of 10 mM NaCl to Mn-stressed durum wheat alleviated Mn-induced damage by reducing Mn accumulation and translocation, while promoting proline accumulation and SOD, APX and GR activities. Irrespective of NaCl level, the combined stress of Mn and NaCl caused more severe oxidative stress, result in further reduction of photosynthetic rate and plant growth in emmer wheat as compared to Mn stress alone. The additively negative effects of NaCl and Mn stress on growth of emmer wheat results from reduced SOD and APX activities as well as Ca, Cu and Fe accumulation in both shoots and roots. These results suggest that salt-tolerant durum wheat is superior to emmer in adapting to Mn stress and the combined stress of salinity and Mn.

     

Ethylenediamine‐N,N′‐disuccinic acid mitigates salt‐stress damages in strawberry by interfering with effects on the plant ionome

This study evaluated the hypothesis that the organic chelant ethylenediamine‐N,N′‐disuccinic acid (EDDS) mitigates plant damage under salinity, and that this is accomplished by EDDS‐induced effects on cation uptake. Damaging effects of salinity on plants often involve inhibited uptake of nutritional cations, such as K and Ca, and excessive accumulation of Na. Therefore, mechanisms that improve uptake of K and Ca, or reduce Na uptake, have a potential for ameliorating salinity damages. Organic chelants increase heavy‐metal cation availability at the site of uptake and increase their uptake by the roots or in planta transport. Although organic chelants are routinely used in agriculture to enhance uptake of heavy‐metal cations into plants, and for soil bioremediation, their effect on uptake of cation‐macronutrients is not known, and neither is their impact on plant function under salinity. In this study, we evaluated the response of strawberry plants to EDDS application (0, 1, 3 and 5 mmol kg soil^?1), under six levels of NaCl (0, 3, 6, 9, 12 and 15 mmol L^?1). EDDS application under salinity improved vegetative development, as well as reproductive growth and chlorophyll content, with statistically significant interaction between chelant dosage and level of salinity. The mitigation of salinity damage by EDDS occurred at high salinity treatments (from 9 mM NaCl). Application rates of 1–3 mmol EDDS kg^?1 were optimal for mitigating salinity effects on reproductive development, but in accordance with the extent of chelant‐induced accumulation of the macronutrients K, Ca and P in the leaves, higher application rates (3–5 mmol EDDS kg^?1) were required for optimal improvement of vegetative development. These results suggest that EDDS improves plant function under mild salinities by interfering with salinity effects on the plant ionome.     

Effects of salinity on O₂ consumption, ROS generation and oxidative stress status of gill mitochondria of the mud crab Scylla serrata

Mitochondrial respiration, activities of electron transport chain enzymes and formation of oxidative stress parameters were investigated in mitochondria isolated from gill tissue of mud crabs (Scylla serrata) as a function of salinity (10 ppt, 17 ppt and 35 ppt). Mitochondrial oxygen consumption rate was higher for succinate as substrate compared with those of glutamate, malate and pyruvate. Complex I and complex II mediated respirations were higher at low salinity (10 ppt) than high salinity (17 ppt and 35 ppt). Although activities of electron transport chain enzymes particularly complexes I (EC 1.6.5.3), II (EC 1.3.99.1) and II-III (EC 1.3.2.1) were elevated linearly in response to salinity treatment, activity of complex V (ATPase, EC 3.6.1.34) was decreased at 35 ppt salinity. However, ATPase activity was higher at 17 ppt salinity in comparison to 10 ppt and 17 ppt salinity. Results of the experiment suggest that high salinity (35 ppt) causes hypoxic state in mitochondria of mud crabs. Hypoxic condition induced by high salinity was accompanied with increased hydrogen peroxide production resulting oxidative stress in mitochondria of crabs. A possible mechanism of hypoxia-induced reactive oxygen species generation and OS due to salinity stress in the crabs is discussed.     

Oxygen consumption of human blood platelets. II. Effect of inhibitors on thrombin-induced oxygen burst

The effect of selected inhibitors on the thrombin-stimulated burst and the basal oxygen consumption of washed human platelets were investigated and compared with inhibition of the release reaction. Cyanide (0.2 mM) caused complete inhibition of the basal respiration, but only 15% inhibition of the thrombin-stimulated burst of oxygen consumption. Similar differential inhibitory effects were observed with oligomycin, antimycin, rotenone and N-ethylmaleimide. Prostaglandin E₁ (0.03 mM) and acetylsalicylic acid (0.8 mM) had little effect on basal respiration, but inhibited the thrombin-stimulated burst of oxygen consumption. N-Ethylmaleimide (0.4 mM) inhibited the release of calcium from platelets by 90%, while prostaglandin E₁, acetylsalicylic acid and the above mitochondrial inhibitors caused no more than 30% inhibition of the release reaction. Our results provide evidence that basal respiration and a portion of the thrombin-stimulated burst of oxygen consumption are involved in respiratory chain phosphorylation, and that this component of the thrombin-stimulated burst may be coupled to the maintenance of the release reaction.