Cytochrome and alternative pathway activity in roots of thermal and non-thermal Agrostis species in response to high soil temperature

Partitioning of respiration between the cytochrome pathway (CP) and the alternative pathway (AP) may play an important role in plant adaptation to extreme environments. We examined changes in partitioning between CP and AP, and viability of roots associated with plant exposure to high soil temperature for two Agrostis species: Agrostis scabra Willd., a species adapted to high-temperature soils in geothermal areas in Yellowstone National Park, and Agrostis stolonifera L. (cv. Penncross) a heat-sensitive grass widely used in cool-climate regions. Roots of A. scabra and A. stolonifera were exposed to soil temperature of 37 or 20°C, while shoots were exposed to 20°C for 28 days. Root viability decreased, and total root respiration increased for both species at 37°C. The decline in root viability and the increase in respiration rates were less pronounced for A. scabra than for A. stolonifera . A larger proportion of total root respiration was attributed to the AP in A. scabra compared with that in A. stolonifera when both species were exposed to 37°C. At 7 and 14 days at 37°C, the relative proportion of respiration passing through AP increased by 12 and 10%, respectively, in A. scabra , whereas in A. stolonifera , AP increased by 4 and 1%, respectively. Our results suggest that maintaining a higher proportion of AP at a high soil temperature may contribute to root thermo-tolerance in A. scabra in comparison with A. stolonifera , and alternative respiration may play an important role in plant adaptation to high soil temperature.     

短日照对休眠诱导期油桃花芽两条电子传递途径的调控

以油桃品种“曙光”为试材,采用呼吸抑制剂法研究了短日照处理下花芽在休眠诱导期两条电子传递途径的发生和运行情况.结果表明:花芽总呼吸速率(Vt)和细胞色素电子传递途径呼吸速率(ρ'Vcyt)均呈双峰曲线变化,短日照可同步诱导两者的一次峰前移、二次峰后延,抑制ρ’Vcyt,但对Vt无显著影响.交替途径容量(Valt)和实际运行活性(ρValt)亦呈双峰曲线,两者基本同步变化,短日照可以显著诱导Valt和ρValt的前期高峰期提前,提高Valt和ρValt,对后期高峰期无明显作用.细胞色素电子传递途径呼吸速率下降和交替途径呼吸速率上升是油桃花芽休眠诱导期的重要特点.从两条电子传递途径的呼吸速率对总呼吸速率的贡献率来看,细胞色素电子传递途径仍是主要电子传递途径,交替途径起辅助与分流作用.     

Activities of respiratory pathways and the pool of nonstructural carbohydrates in greening leaves of spring wheat seedlings

Seedlings of spring wheat (Triticum aestivum L.) were used to study the dynamics of leaf respiration, the respiratory pathway ratio, and relation of activities of these pathways to the content of soluble carbohydrates in the leaf during greening of seedlings for 48 h under continuous photosynthetically active light (190 μmol/(m² s)). Changes in leaf respiration during de-etiolation were closely related to modulation of the alternative respiratory pathway (AP) activity. The rate of cytochrome respiratory pathway (CP) depended directly on the carbohydrate content and growth rate. These relations suggest that the substrate regulation of CP activity during greening is mediated by the energy needs for growth and is effectively regulated by the mechanism of respiratory control. The highest rates of AP were observed after a 6-h exposure of seedlings to light. The proportion of CP/AP at this stage was close to unity. The temporal pattern of AP activity during de-etiolation was independent on the content of soluble carbohydrates. Hence, in addition to substrate regulation of AP, there are other intricate mechanisms of AP involvement. Our results are in accordance to the state that the alternative respiratory pathway participates in maintaining homeostasis in phototrophic cells during development of the photosynthetic function.     

The Alternative Respiration Pathway in Leaves ofRhodiola roseaand Ajuga reptans: Presumable Physiological Role

The effect of growth conditions and plant age on the relationships between respiratory pathways was investigated in Rhodiola roseaand Ajuga reptans.The alternative pathway (AP) contributed 0–50% to the leaf respiration; however, this pathway was absent from the overwintered leaves of A. reptans.In both plant species, AP contributed 15–20% to the respiration of mature leaves, and in the young rapidly expanding leaves the contribution was twice higher. The highest AP contribution (40–50%) was found in the leaves of A. reptansplants grown in an experimental plot in full light. As compared to the plot-grown plants, A. reptansplants grown in their natural habitats were characterized by a lower AP contribution to the respiration of leaves; they contained two times less nonstructural carbohydrates and accumulated less biomass. We conclude that a high AP contribution to the respiration of leaves correlates with their rapid growth and that a high supply of respiratory substrates is one of prerequisite for the AP activation.     

Salinity tolerance is related to cyanide‐resistant alternative respiration in Medicago truncatula under sudden severe stress

Salt respiration is defined as the increase of respiration under early salt stress. However, the response of respiration varies depending on the degree of salt tolerance and salt stress. It has been hypothesized that the activity of the alternative pathway may increase preventing over‐reduction of the ubiquinone pool in response to salinity, which in turn can increase respiration. Three genotypes of Medicago truncatula are reputed as differently responsive to salinity: TN1.11, A17 and TN6.18. We used the oxygen‐isotope fractionation technique to study the in vivo respiratory activities of the cytochrome oxidase pathway (COP) and the alternative oxidase pathway (AOP) in leaves and roots of these genotypes treated with severe salt stress (300 mM) during 1 and 3 days. In parallel, AOX capacity, gas exchange measurements, relative water content and metabolomics were determined in control and treated plants. Our study shows for first time that salt respiration is induced by the triggered AOP in response to salinity. Moreover, this phenomenon coincides with increased levels of metabolites such as amino and organic acids, and is shown to be related with higher photosynthetic rate and water content in TN6.18.     

Light induction of alternative pathway capacity in leaf slices of Belgium endive

The effect of light on the development of the capacity for alternative pathway respiration was investigated in leaf slices of Belgium endive (Cichorum intybus L. cv. deliva). Dark-grown plants possessed little capacity for the cyanide-insensitive alternative pathway. In contrast, plants grown in continuous light had significant alternative pathway capacity. Light-grown plants also had substantially higher concentrations of ethanol-soluble carbohydrates in their leaves than plants grown in complete darkness. Despite these differences in leaf carbohydrate status and alternative pathway capacity of light- and dark-grown leaf tissue, no differences were found in the activity of the alternative pathway, which was negligible in both treatments. Dark-grown plants were adenylate restricted, as indicated by the increase in cytochrome pathway activity following uncoupling. Adenylates did not limit respiration in light-grown leaf tissue. Plants that had been grown for 8d in complete darkness were also transferred to continuous light. Respiration of dark controls steadily declined over 11d following the transfer of plants to the light, due primarily to a decrease in cytochrome pathway activity. No such decline was observed in the plants transferred to continuous light. Transfer to continuous light led to significant increases in alternative pathway capacity relative to the dark controls. Alternative pathway activity remained negligible in both the dark controls and in plants transferred to continuous light. The results of this study suggest then that light per se may be responsible for the induction of alternative pathway capacity in Belgium endive.     

Overexpression of wheat alternative oxidase gene Waox1a alters respiration capacity and response to reactive oxygen species under low temperature in transgenic Arabidopsis

Under low temperature conditions, the cytochrome pathway of respiration is repressed and reactive oxygen species (ROS) are produced in plants. Mitochondrial alternative oxidase (AOX) is the terminal oxidase responsible for the cyanide-insensitive and salicylhydroxamic acid-sensitive respiration. To study functions of wheat AOX genes under low temperature, we produced transgenic Arabidopsis by introducing Waox1a expressed under control of the cauliflower mosaic virus (CaMV) 35S promoter in Arabidopsis thaliana. The enhancement of endogenous AOX1a expression via low temperature stress was delayed in the transgenic Arabidopsis. Recovery of the total respiration activity under low temperature occurred more rapidly in the transgenic plants than in the wild-type plants due to a constitutively increased alternative pathway capacity. Levels of ROS decreased in the transgenic plants under low temperature stress. These results support the hypothesis that AOX alleviates oxidative stress when the cytochrome pathway of respiration is inhibited under abiotic stress conditions.     

Response of mitochondrial antioxidant system and respiratory pathways to reactive nitrogen species in pea leaves

Nitric oxide (NO) has emerged as an important signaling molecule in plants, but little is known about the effects of reactive nitrogen species in plant mitochondria. In this study, the effects of DETA‐NONOate, a pure NO slow generator, and of SIN‐1 (3‐morpholinosydnonimine), a peroxynitrite producer, on the activities of respiratory pathways, enzymatic and non‐enzymatic antioxidants have been investigated in isolated mitochondria from pea leaves. No significant changes in lipid peroxidation, protein oxidation or in ascorbate and glutathione redox state were observed after DETA‐NONOate treatments whereas cytochrome pathway (CP) respiration was reversibly inhibited and alternative pathway (AP) respiration showed little inhibition. On the other hand, NO did not affect neither activities of Mn superoxide dismutase (Mn‐SOD) nor enzymes involved in the ascorbate and glutathione regeneration in mitochondria except for ascorbate peroxidase (APX), which was reversely inhibited depending on ascorbate concentration. Finally, SIN‐1 treatment of mitochondria produced a decrease in CP respiration, an increase in protein oxidation and strongly inhibited APX activity (90%), with glutathione reductase and dehydroascorbate reductase (DHAR) being moderately inhibited (30 and 20%, respectively). This treatment did not affect monodehydroascorbate reductase (MDHAR) and Mn‐SOD activities. Results showed that mitochondrial nitrosative stress was not necessarily accompanied by oxidative stress. We suggest that NO‐resistant AP and mitochondrial APX may be important components of the H₂O₂‐signaling pathways under nitrosative stress induced by NO in this organelle. Also, MDHAR and DHAR, via ascorbate regeneration, could constitute an essential antioxidant defense together with Mn‐SOD, against NO and ONOO^? stress in plant mitochondria.     

Leaf respiration and alternative oxidase in field-grown alpine grasses respond to natural changes in temperature and light

? We report the first investigation of changes in electron partitioning via the alternative respiratory pathway (AP) and alternative oxidase (AOX) protein abundance in field-grown plants and their role in seasonal acclimation of respiration. ? We sampled two alpine grasses native to New Zealand, Chionochloa rubra and Chionochloa pallens, from field sites of different altitudes, over 1 yr and also intensively over a 2-wk period. ? In both species, respiration acclimated to seasonal changes in temperature through changes in basal capacity (R??) but not temperature sensitivity (E?). In C. pallens, acclimation of respiration may be associated with a higher AOX : cytochrome c oxidase (COX) protein abundance ratio. Oxygen isotope discrimination (D), which reflects relative changes in AP electron partitioning, correlated positively with daily integrated photosynthetically active radiation (PAR) in both species over seasonal timescales. Respiratory parameters, the AOX : COX protein ratio and D were stable over a 2-wk period, during which significant temperature changes were experienced in the field. ? We conclude that respiration in Chionochloa spp. acclimates strongly to seasonal, but not to short-term, temperature variation. Alternative oxidase appears to be involved in the plant response to both seasonal changes in temperature and daily changes in light, highlighting the complexity of the function of AOX in the field.     

Effects of light on respiration and oxygen isotope fractionation in soybean cotyledons

Light effects on electron flow through the cyanide-resistant respiratory pathway, oxygen isotope fractionation and total respiration were studied in soybean (Glycine max L.) cotyledons. During the first 12 h of illumination there was an increase in both electron partitioning through the alternative pathway and oxygen isotope fractionation by the alternative oxidase. The latter probably indicates a change in the properties of the alternative oxidase. There was no engagement of the alternative oxidase in darkness and its fractionation was 27‰. In green cotyledons 60% of the respiration flux was through the alternative pathway and the alternative oxidase fractionation was 32‰. Exposing previously illuminated tissue to continuous darkness induced a decrease in the electron partitioning through the alternative pathway. However, this decrease was not directly linked with the low cellular sugar concentration resulting from the lack of light because 5 min of light every 12 h was sufficient to keep the alternative pathway engaged to the same extent as plants grown under control conditions.     

Effects of water stress on development, operation and gene expression of cyanide-resistant respiratory pathway in wheat

Osmotic dehydration of wheat seedlings in-0.5 MPa polyethylene glycol (PEG) solutions for 24, 48 and 72 h resulted in mild, moderate and severe water stress respectively in leaves, but only caused mild water stress in roots as reflected by the changes in relative water content (RWC). In response to the above water stress conditions, leaf total respiratory rate (V_t) decreased progressively, and the alternative pathway (AP) capacity (V_(alt)) and its actual operation activity (ρV_(alt)) decreased more severely. Water stress also led to continuous reduction in cytochrome pathway (CP) activity ((ρ' V_(cyt)) and different changes in the contribution of ρV_(alt) and ρ' V_(cyt) to V_t in leaves, with ρV_(alt)/V_t decreasing and ρ' V_(cyt)/V_t increasing. The change pattern of root V_t was similar to that of its RWC, while root V_(alt) and ρV_(alt) were found to decrease during the first 24 h of stress and thereafter recover to a level close to that of the control (O h). These data indicate that the alt     

Energy balance of a plant under normal and unfavourable conditions

Two main components of plant energy balance are analyzed--photosynthesis and dark respiration. Different plant species, growing in optimal conditions and differing in photosynthetic metabolism, productivity and potential resistance to stress, have constant ratio between total dark respiration and grossphotosynthesis. The ratio about 38-40% at the phase of active growth. This value reflect plant state, when income (assimilation) is maximized, and expense (total oxidation) is minimized. Intensities of dark respiration of plants in darkness and light are similar despite the fact that the plants have considerable differences: 1) different carbon sources--young assimilates in light and reserve ones in the dark; 2) biochemical modifications (or inhibition) of some stages of dark respiration in light; 3) intensification of alternative respiration in unfavourable conditions differing in dark and light variants. Ratio between intensity of dark respiration and photosynthesis increases in plants growing in unfavourable conditions. This increase is more considerable in plants that are less resistant to the given stress. Characters of energy balance can be used for estimation of physiological status of plants, prediction of resistance and potential productivity at seedling stage.     

Effect of cadmium on growth and respiration of barley plants grown under two temperature regimes

We studied cadmium effect on the respiratory pathways ratio in the organs of barley (Hordeum distichum L., cv. Novichok) plants grown in water culture at two temperature regimes. Mineral nutrients were supplied daily in exponentially increasing amounts in order to provide for steady-state growth. CdSO₄ (30, 60, or 100 μmol/l) was added to nutrient solution at a single time in the beginning of the exponential growth period (19 days after germination). In further 6 days, the relative growth rate and biomass accumulation declined stronger with the increase in the cadmium concentration in plants grown at 13/8°C (day/night) than at 21/17°C (day/night). Cadmium suppressed root respiration (down to 60% of control) stronger than leaf respiration, and the roots manifested a higher sensitivity to the inhibitor of alternative oxidase, salicylhydroxamic acid. The respiratory pathways ratio in the roots occurred against the background of activated lipid peroxidation (POL). The highest POL activity and the highest proportion of alternative respiration pathway (AP) (up to 46% of total respiration) were observed in the roots in the presence of the highest cadmium concentration (100 μM) under lower temperature (13/8°C). Thus, high cadmium concentrations affected strongly the total rate of respiration and respiratory pathways ratio. Growth temperature modulated Cd effects on respiration. AP activation is one of the mechanisms for maintenance of root cell homeostasis under cadmium-induced stress.     

Salicylic acid enhances the activity of the alternative pathway of respiration in tobacco leaves and induces thermogenicity

A rise in the level of endogenous salicylic acid (SA) during flowering of the thermogenic voodoo lily, Sauromatum guttatum, leads to a pronounced temperature elevation by stimulation of the alternative respiratory pathway. We have studied the thermal response of tobacco (Nicotiana tabacum L.) leaves, a non-thermogenic tissue, to exogenous SA, and its relation to alternative respiration. A reproducible increase in surface temperature of 0.5–1.0°C was registered with high-resolution infrared cameras. The same phenomenon was observed when 2,6-dihydroxybenzoic acid, an active analogue of SA, was used. Non-active SA analogues, such as 3- and 4-hydroxybenzoic acid, did not induce thermogenicity. The thermal effect of SA was abolished with inhibitors of the alternative pathway, such as salicylhydroxamic acid and propyl gallate. Polarographic measurement of the respiratory activity, including that of the alternative pathway in SA-treated plants, showed a significant increase of both total respiration and the alternative pathway compared with non-treated controls. Therefore, we postulate that, as in thermogenic species, SA enhances the activity of total respiration and of the cyanide-resistant pathway in tobacco leaves, subsequently leading to an elevation in surface temperature.     

The rate of development of water deficits affects Saxifraga cernua leaf respiration

We allowed plant water deficits to develop at two different rates following the cessation of watering in order to investigate the effects of water stress on cytochrome pathway and alternative pathway respiration in the leaves of the arctic herb Saxifraga cernua. Plants were pretreated by growth in either a commercial organic (CO) mixture or a vermiculite-perlite (VP) mixture, which allowed the complete development of water deficits in 19 and 8 days, respectively. The rate of water potential reduction was approximately 0.11 MPa day⁻¹ in the leaves of CO plants, compared to a reduction of 0.21 MPa day⁻¹ in leaves of VP plants. Osmotic adjustment occurred to a greater extent in leaves of CO plants and corresponded with an increase in ethanol-soluble sugars. In leaves of CO plants, cytochrome pathway activity gradually declined from that of control rates until day 11, and then declined more rapidly. In contrast, cytochrome pathway activity significantly increased in response to water deficits in leaves of VP plants. In leaves of both CO and VP plants, alternative pathway activity declined as water stress progressed. Relatively severe water deficits reduced alternative pathway capacity in leaves of both CO and VP plants. We also investigated the effect of previous exposure to water deficits on leaf respiration. In plants that had previously experienced three cycles of water stress, the increase in cytochrome pathway activity during the fourth water stress cycle was small compared to the increase observed in leaves of plants experiencing water stress for the first time. These results suggest that cytochrome pathway activity is differentially sensitive to the rate of development of plant water deficits and that respiratory responses to acute water stress are not necessarily similar to the responses to chronic water stress.     

Physiological Characteristics and Alternative Respiratory Pathway under Salt Stress in Two Wheat Cultivars Differing in Salt Tolerance

Changes in respiratory pathway, dry weight, contents of proline, ATP, Na⁺and K⁺were investigated under five salinity treatments in the leaves of plants of spring wheat (Triticum aestivumL.). Two cultivars differing in salt resistance, namely, 89122 (salt-tolerant) and Longchun13 (salt-sensitive), were used. The decrease in dry weight and K⁺content was observed with the increasing NaCl concentrations, but more in cv. 89122 plants than in Longchun13 plants. Contents of proline and Na⁺in both cvs increased greatly, but the former increased more in 89122 while the latter more in Longchun13 plants. In all salinity treatments tested, a salt-induced increase in the activity of the alternative pathway was found, although cytochrome pathway (CP) still remained the main electron transport pathway. ATP production changed in parallel with CP operation. Cv. 89122 plants could produce more ATP than cv. Longchun13 plants exposed to each salinity treatment and their ATP generation could even be stimulated in contrast to its rapidly decline in Longchun13 plants with increased salinity stress. The possible relationship between respiration metabolism and above mentioned physiological changes is discussed.     

小麦幼苗叶片抗氰呼吸对轻度水分胁迫的响应

小麦幼苗经－０．５ＭＰa聚乙二醇（ＰＥＧ－６０００）溶液暗中渗透迫０、６、１２、１８、２４、３０、３６、４２、４８h,叶片的总呼吸速率（Ｖt)呈现先上升后降低的趋势,交替途径呼吸也表现出相同的变化模式。水分胁迫初期（０－１２h),交替途径容量（Ｖalt)、实际运行活性（ρValt)及运行系数（ρ值）均上升,此后（１８－４８h）逐渐下降,水分胁迫也影响了呼吸电子流在２条呼吸途径中的分配比例,胁迫初期的０－１２h内,流经交替途径的电子流增多,而流向细胞色素主路的电子流减少,但随着胁迫时间的延长,交替途径的贡献降低,而细胞色素主路的贡献增加,说明小麦叶片的抗氰呼吸在水分胁迫初期被诱导增加,而随着胁迫进行的延长又表现为下降。