Cytochrome Pathway Respiration Research Articles

Possible involvement of nitric oxide in promoting the initial growth of rice seedlings at low temperature by inoculation of Bacillus pumilus strain TUAT1 spores

ABSTRACT Bacillus pumilus strain TUAT1, a type of plant growth – promoting rhizobacteria (PGPR), is used as an ingredient in biofertilizer. Although we confirmed that B. pumilus TUAT1 promotes the growth of rice seedlings at 25°C, this effect has not been shown at lower temperatures. In this study, we confirmed that inoculation of rice seedlings with spores of B. pumilus TUAT1 promoted seedling emergence from soil and subsequent growth at 15°C. Except for the effect on root growth, these effects disappeared when seeds were treated with 2–4-carboxyphenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide, a nitric oxide (NO) scavenger, simultaneously with B. pumilus TUAT1 inoculation. Increased NO accumulation was detected in seed embryos 3 h after inoculation, suggesting that NO plays a role in the effects of B. pumilus TUAT1 inoculation. Transcriptome analysis using RNA-seq suggested the involvement of immune responses and cytochrome respiratory pathways in increasing NO levels in embryos after inoculation with B. pumilus TUAT1. Transcriptome analysis also indicated that transcription of genes involved in cold tolerance was accelerated in embryos following the increase in NO resulting from inoculation with B. pumilus TUAT1. These findings suggest that a novel NO signaling mechanism is involved in PGPR-induced growth promotion in plants. These results also indicate that inoculation with B. pumilus TUAT1 spores may help overcome the initial growth failure of seedlings in direct sowing culture of rice in cold regions.

Developmentally regulated mitochondrial biogenesis and cell death competence in maize pollen

BackgroundCytoplasmic male sterility (CMS) is a maternally inherited failure to produce functional pollen that most commonly results from expression of novel, chimeric mitochondrial genes. In Zea mays, cytoplasmic male sterility type S (CMS-S) is characterized by the collapse of immature, bi-cellular pollen. Molecular and cellular features of developing CMS-S and normal (N) cytoplasm pollen were compared to determine the role of mitochondria in these differing developmental fates.ResultsTerminal deoxynucleotidyl transferase dUTP nick end labeling revealed both chromatin and nuclear fragmentation in the collapsed CMS-S pollen, demonstrating a programmed cell death (PCD) event sharing morphological features with mitochondria-signaled apoptosis in animals. Maize plants expressing mitochondria-targeted green fluorescent protein (GFP) demonstrated dynamic changes in mitochondrial morphology and association with actin filaments through the course of N-cytoplasm pollen development, whereas mitochondrial targeting of GFP was lost and actin filaments were disorganized in developing CMS-S pollen. Immunoblotting revealed significant developmental regulation of mitochondrial biogenesis in both CMS-S and N mito-types. Nuclear and mitochondrial genome encoded components of the cytochrome respiratory pathway and ATP synthase were of low abundance at the microspore stage, but microspores accumulated abundant nuclear-encoded alternative oxidase (AOX). Cytochrome pathway and ATP synthase components accumulated whereas AOX levels declined during the maturation of N bi-cellular pollen. Increased abundance of cytochrome pathway components and declining AOX also characterized collapsed CMS-S pollen. The accumulation and robust RNA editing of mitochondrial transcripts implicated translational or post-translational control for the developmentally regulated accumulation of mitochondria-encoded proteins in both mito-types.ConclusionsCMS-S pollen collapse is a PCD event coincident with developmentally programmed mitochondrial events including the accumulation of mitochondrial respiratory proteins and declining protection against mitochondrial generation of reactive oxygen species.

Seed germination, respiratory processes and phosphatidic acid accumulation in Arabidopsis diacylglycerol kinase knockouts – The effect of brassinosteroid, brassinazole and salinity

Using Arabidopsis thaliana wild type (WT) plants and diacylglycerol kinase knockouts (single mutants - dgk3, dgk1, dgk6; double mutants - dgk3dgk7, dgk5dgk6, dgk1dgk2) we observed that the inhibitor of brassinosteroid (BR) biosynthesis, brassinazole (BRZ), drastically decreased germination of dgk mutants under salt stress, while BRZ co-administration with 24-epibrassinolide (EBL) partially improved germination rates. We also observed a statistically significant decrease in alternative and cytochrome respiratory pathways in response to BRZ treatment under salinity conditions. We showed that production of the lipid second messenger phosphatidic acid (PA) is impaired in dgk mutants in response to EBL treatment and inhibitor of diacylglycerol kinase (DGK) – R59022. This study demonstrates that dgk mutants possess lower germination rates, lower total respiration rates, an alternative respiratory pathway and PA content under optimal and high salinity conditions in response to EBL treatment comparing to WT plants.

Bud dormancy breaking affects respiration and energy balance of bilberry shoots in the initial stage of growth

Respiration and heat production in the shoots of bilberry (Vaccinium myrtillus L.) were studied at the beginning of growth after breaking bud dormancy by means of transfer of the shoots to indoor conditions (November–April) and upon natural sprouting in spring (May). The buds released from dormancy at the beginning of winter sprouted slower and showed lower respiratory activity than the buds that started growing in May. In May, cytochrome respiratory pathway in sprouting buds was 1.3 times more active than energetically ineffective alternative pathway, whereas activity of cytochrome pathway in December was 1.4 times lower as compared with the alternative. In November–December, the rate of heat evolution by the buds was 3–5 times lower than in April–May. In case of early breaking of bud dormancy, the share of respiration energy dissipated as heat was 30% on average. In the buds whose growth was induced later, the value of this parameter was twice as much. The ratio between heat evolution and respiration depended on temperature. High temperature more intensely activated heat evolution than respiration, which caused a decrease in the level of metabolic energy available for growth. In the temperature range of 5–15°C characteristic of the beginning of vegetation, the share of respiration energy dissipated as heat was 2–3 times lower than at 20–30°C, which reflects a great adaptability of V. myrtillus to climatic conditions of the region. Our data suggest that progression through a full cycle of winter dormancy is physiologically important for shoot growth. Early dormancy release brought about changes in respiration and energy balance of the shoots in the initial stage of extra-bud growth.

Improved photosynthetic performance during severe drought in Nicotiana tabacum overexpressing a nonenergy conserving respiratory electron sink.

Chloroplasts have means to manage excess reducing power but these mechanisms may become restricted by rates of ATP turnover. Alternative oxidase (AOX) is a mitochondrial terminal oxidase that uncouples the consumption of reducing power from ATP synthesis. Physiological and biochemical analyses were used to compare respiration and photosynthesis of Nicotiana tabacum wild-type (WT) plants with that of transgenic lines overexpressing AOX, under both well-watered and drought stress conditions. With increasing drought severity, AOX overexpression acted to increase respiration in the light (RL ) relative to WT. CO2 and light response curves indicated that overexpression also improved photosynthetic performance relative to WT, as drought severity increased. This was not due to an effect of AOX amount on leaf water status or the development of the diffusive limitations that occur due to drought. Rather, AOX overexpression dampened photosystem stoichiometry adjustments and losses of key photosynthetic components that occurred in WT. The results indicate that AOX amount influences RL , particularly during severe drought, when cytochrome pathway respiration may become increasingly restricted. This impacts the chloroplast redox state, influencing how the photosynthetic apparatus responds to increasing drought severity. In particular, the development of biochemical limitations to photosynthesis are dampened in plants with increased nonenergy conserving RL .

Effects of chlorine dioxide treatment on respiration rate and ethylene synthesis of postharvest tomato fruit

Tomato fruit at the mature green stage were treated with ClO2 gas in a sealed container for 12h, and then stored at 23°C with 85% relative humidity (RH) for 23 d. Respiration rate, respiration-related enzymes including phosphohexose isomerase (PHI), succinate dehydrogenase (SDH), and glucose-6-phosphate dehydrogenase (G-6-PDH) and 6-phosphogluconate dehydrogenase (6-PGDH), ethylene production, and the expression of LeAOX1a, LeCOX1, LeACS2, LeACS4 and LeACO1 genes were measured. The results showed that application of ClO2 gas was effective in reducing total respiration, cytochrome pathway respiration and the expression of LeCOX1, but no significant reduction in the activities of respiration-related enzymes was observed during storage. Fruit treated with ClO2 resulted in lower ethylene production. Furthermore, the expression of ethylene biosynthesis related genes, including LeACS2, LeACS4 and LeACO1 was reduced by the ClO2 treatment. These results indicate that ClO2 treatment might delay the ripening of tomato fruit, possibly by a mechanism involving suppression of respiration rate and ethylene biosynthesis.

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.

Evidence of an Alternative Oxidase Pathway for Mitochondrial Respiration in the Scuticociliate Philasterides dicentrarchi

The presence of an alternative oxidase (AOX) in the mitochondria of the scuticociliate P. dicentrarchi was investigated. The mitochondrial oxygen consumption was measured in the presence of KCN, an inhibitor of cytochrome pathway (CP) respiration and salicylhydroxamic acid (SHAM), a specific inhibitor of alternative pathway (AP) respiration. AOX expression was monitored by western blotting with an AOX polyclonal antibody. The results showed that P. dicentrarchi possesses a branched mitochondrial electron transport chain with both cyanide-sensitive and -insensitive oxygen consumption. Mitochondrial respiration was partially inhibited by cyanide and completely inhibited by the combination of cyanide and SHAM, which is direct evidence for the existence of an AP in this ciliate. SHAM significantly inhibited in vitro growth of trophozoites both under normoxic and hypoxic conditions. AOX is a 42kD monomeric protein inducible by hypoxic conditions in experimental infections and by CP inhibitors such as cyanide and antimycin A, or by AP inhibitors such as SHAM. CP respiration was greatly stimulated during the exponential growth phase, while AP respiration increased during the stationary phase, in which AOX expression is induced. As the host does not possess AOX, and because during infection P. dicentrarchi respires via AP, it may be possible to develop inhibitors targeting the AP as a novel anti-scuticociliate therapy.

Chlorine dioxide treatment decreases respiration and ethylene synthesis in fresh‐cut ‘Hami’ melon fruit

SummaryThe effects of chlorine dioxide (ClO2) on respiration and ethylene synthesis of fresh‐cut melon fruit and the possible mechanisms involved were investigated. Fresh‐cut ‘Hami’ melon fruit fumigated with gas ClO2 in sealed container for 12 h and then stored at 5 °C with 95% relative humidity (RH) for 19 days. Results showed that fruit treated with ClO2 resulted in lower rates of the total respiration, alternative pathway respiration, cytochrome pathway respiration and ethylene production. Furthermore, the expressions of ethylene biosynthesis‐related genes, including CmACS2, CmACO1 and CmACO3 were reduced by ClO2 treatment. Taken together, it is suggested that ClO2 treatment might be an effective way to delay ripening of fresh‐cut ‘Hami’ melon, partially due to the reduced respiration and ethylene biosynthesis.

Comparison of Intact Arabidopsis thaliana Leaf Transcript Profiles during Treatment with Inhibitors of Mitochondrial Electron Transport and TCA Cycle

Plant mitochondria signal to the nucleus leading to altered transcription of nuclear genes by a process called mitochondrial retrograde regulation (MRR). MRR is implicated in metabolic homeostasis and responses to stress conditions. Mitochondrial reactive oxygen species (mtROS) are a MRR signaling component, but whether all MRR requires ROS is not established. Inhibition of the cytochrome respiratory pathway by antimycin A (AA) or the TCA cycle by monofluoroacetate (MFA), each of which initiates MRR, can increase ROS production in some plant cells. We found that for AA and MFA applied to leaves of soil-grown Arabidopsis thaliana plants, ROS production increased with AA, but not with MFA, allowing comparison of transcript profiles under different ROS conditions during MRR. Variation in transcript accumulation over time for eight nuclear encoded mitochondrial protein genes suggested operation of both common and distinct signaling pathways between the two treatments. Consequences of mitochondrial perturbations for the whole transcriptome were examined by microarray analyses. Expression of 1316 and 606 genes was altered by AA and MFA, respectively. A subset of genes was similarly affected by both treatments, including genes encoding photosynthesis-related proteins. MFA treatment resulted in more down-regulation. Functional gene category (MapMan) and cluster analyses showed that genes with expression levels affected by perturbation from AA or MFA inhibition were most similarly affected by biotic stresses such as pathogens. Overall, the data provide further evidence for the presence of mtROS-independent MRR signaling, and support the proposed involvement of MRR and mitochondrial function in plant responses to biotic stress.

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 H(2) O(2) -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.

Photosynthesis, respiration and antioxidant enzymes in pepper leaves under drought and heat stresses

Leaf gas exchange, chlorophyll fluorescence and activities of antioxidant enzymes were studied in two pepper (Capsicum annuum L.) cultivars grown under drought (D) and heat (H), as well as under drought and heat in combination (HD). The drought-tolerant cv. Zhengjiao 13 exhibited greater net photosynthetic rate (PN) and cytochrome respiratory pathway activity (RSHAM), and lower contents of superoxide radical and hydrogen peroxide, as compared to the drought-sensitive cv. Longkouzaojiao. The PN and RSHAM decreased and ROS production increased under D and HD in both cultivars. As compared to the Longkouzaojiao, Zhengjiao 13 retained higher non-photochemical quenching (NPQ), photorespiration rate (RL), and alternative respiratory pathways (RKCN) under D and HD. Drought increased the superoxide dismutase (SOD) and ascorbate peroxidase (APX) activities in the cytosol, chloroplasts and mitochondria in the two cultivars. Although SOD and APX activities decreased in Longkouzaojiao under HD, SOD activity increased in Zhengjiao 13. There was no H-induced reactive oxygen species production due to increase of RL, NPQ, RSHAM, RKCN and activities of SOD and APX. However, H slightly decreased the PN. The results indicated that HD was more detrimental than either stress alone.

Deletion of the mitochondrial NADH kinase increases mitochondrial DNA stability and life span in the filamentous fungus Podospora anserina

In the filamentous fungus Podospora anserina, aging is systematically associated with mitochondrial DNA (mtDNA) instability. A causal link between deficiency of the cytochrome respiratory pathway and lifespan extension has been demonstrated. Knock out of the cytochrome respiratory pathway induces the expression of an alternative oxidase and is associated with a reduction in free radical production. The question of the links between mtDNA stability, ROS generation and lifespan is therefore clearly raised in this organism. NADPH lies at the heart of many anti-oxidant defenses of the cell. In Saccharomyces cerevisiae, the mitochondrial NADPH is largely provided by the Pos5 NADH kinase. We show here that disruption of PaNdk1 encoding the potential mitochondrial NADH kinase of P. anserina leads to severe somatic and sexual defects and to hypersensitivity to hydrogen peroxide and paraquat. Surprisingly, it also leads to a spectacular increase of mtDNA stability and lifespan. We propose that an adaptative metabolic change including the induction of the alternative oxidase can account for these results.

Interference BetweenTobacco necrosis virusandTurnip crinkle virusinNicotiana benthamiana

Mixed infections of Nicotiana benthamiana plants by Tobacco necrosis virus (TNV) and Turnip crinkle virus (TCV) exhibited an interference interaction. Accumulation of TNV (+)RNA as well as capsid protein in mixed infection were considerably lower than that of singly infected plants. There were also a slight reduction in the levels of TCV (+)RNA and capsid protein in doubly infected plants, which displayed the concentration of both viruses decreased in dually infected plants. Tissue immunoblot analysis of systemic N. benthamiana leaves infected by TNV and TCV singly or doubly showed the interference between the two viruses in situ, which exhibited the decrease of both viruses in doubly infected leaves although the distribution of them did not change remarkably. These results were consistent with the hybridization analysis of viral genomic RNA and coat protein. Both cross-protection test and mixed infection of the two viruses confirmed TCV had relatively stronger interference to the infection of TNV. Interference infection by TNV and TCV induced higher increase in the levels of cytochrome pathway respiration and alternative pathway respiration in host plants, especially the latter. Interference often occurred in different strains of one kind of virus or two different closely related viruses in one genus. Our results showed that interference could also occur in different viruses belonging to different genera.

Acanthamoeba alternative oxidase genes: Identification, characterisation and potential as antimicrobial targets

Alternative oxidase (AOX) is a mitochondrial protein that acts as an alternative terminal oxidase to the conventional cytochrome oxidases. It is present in certain prokaryotes, plants, fungi and some protozoa but absent in mammals. AOX activity has previously been described in Acanthamoeba, although no genetic evidence has been reported. Herein, two AOX ( AcAOX) genes designated isoforms A and B, were obtained from Acanthamoeba castellanii by a combination of degenerate PCR from cDNA and a series of 5′ and 3′ rapid amplification of cDNA ends. The corresponding genomic sequences of these AcAOXs were also obtained. Each gene spans six exons over a region of 1607 and 1619 bp, respectively. Isoforms A and B have open reading frames of 1113 and 1125 bp, respectively. Each encodes a protein with a predicted molecular weight of 42 kDa. Each AcAOX protein has a predicted cleavable mitochondrial targeting sequence. The full-length AcAOX is functionally active as it complements hemL-deficient Escherichia coli and inhibited by the inhibitor of AOX, salicylhydroxamic acid (SHAM). SHAM is effective against A. castellanii and Acanthamoeba polyphaga only when used in conjunction with antimycin A, an inhibitor of the conventional cytochrome respiratory pathway. Transcripts for AcAOX are increased during the encystment process, indicating a possible role for alternative respiration during stress.

Changes in electron transport, superoxide dismutase and ascorbate peroxidase isoenzymes in chloroplasts and mitochondria of cucumber leaves as influenced by chilling

In order to clarify the relationship between chill-induced disturbance in photosynthetic, respiratory electron transport and the metabolism of reactive oxygen species (ROS), leaf gas exchange, chlorophyll fluorescence quenching, respiration, and activities of superoxide dismutase (SOD) and ascorbate peroxidase (APX) were investigated in chloroplasts and mitochondria of cucumber (Cucumis sativus) leaves subjected to a chill (8 °C) for 4 d. Chilling decreased net photosynthetic rate (PN) and quantum efficiency of photosystem 2 (ΦPS2), but increased the ratio of ΦPS2 to the quantum efficiency of CO2 fixation (ΦCO2) and non-photochemical quenching (NPQ) in cucumber leaves. While chilling inhibited the activity of cytochrome respiration pathway, it induced an increase of alternative respiration pathway activity and the reduction level of Q-pool. Chilling also significantly increased O2• production rate, H2O2 content, and SOD and APX activities in chloroplasts and mitochondria. There was a more significant increase in SOD and APX activities in chloroplasts than in mitochondria with the increase of membrane-bound Fe-SOD and tAPX in chloroplasts being more significant than other isoenzymes. Taken together, chilling inhibited PN and cytochrome respiratory pathway but enhanced the photosynthetic electron flux to O2 and over-reduction of respiratory electron transport chain, resulting in ROS accumulation in cucumber leaves. Meanwhile, chilling resulted in an enhancement of the protective mechanisms such as thermal dissipation, alternative respiratory pathway, and ROS-scavenging mechanisms (SODs and APXs) in chloroplasts and mitochondria.

Contribution of the alternative pathway to respiration during thermogenesis in flowers of the sacred lotus.

We report results from in vivo measurements, using oxygen isotope discrimination techniques, of fluxes through the alternative and cytochrome respiratory pathways in thermogenic plant tissue, the floral receptacle of the sacred lotus (Nelumbo nucifera). Fluxes through both pathways were measured in thermoregulating flowers undergoing varying degrees of thermogenesis in response to ambient temperature. Significant increases in alternative pathway flux were found in lotus receptacles with temperatures 16 degrees C to 20 degrees C above ambient, but not in those with lesser amounts of heating. Alternative pathway flux in the hottest receptacles was 75% of the total respiratory flux. In contrast, fluxes through the cytochrome pathway did not change significantly during thermogenesis. These data support the hypothesis that increased flux through the alternative pathway is responsible for heating in the lotus and that it is unlikely that uncoupling proteins, which would have produced increased fluxes through the cytochrome pathway, contribute significantly to heating in this tissue. Comparisons of actual flux, with capacity determined using inhibitors, suggested that the alternative pathway was operating at close to maximum capacity in heating tissues of lotus. However, in nonheating tissues the inhibitor data significantly overestimated the alternative pathway flux. This confirms that isotopic measurements are necessary for accurate determination of fluxes through the two pathways.

Interaction Between the oxa1 and rmp1 Genes Modulates Respiratory Complex Assembly and Life Span in Podospora anserina

A causal link between deficiency of the cytochrome respiratory pathway and life span was previously shown in the filamentous fungus Podospora anserina. To gain more insight into the relationship between mitochondrial function and life span, we have constructed a strain carrying a thermosensitive mutation of the gene oxa1. OXA1 is a membrane protein conserved from bacteria to human. The mitochondrial OXA1 protein is involved in the assembly/insertion of several respiratory complexes. We show here that oxa1 is an essential gene in P. anserina. The oxa1(ts) mutant exhibits severe defects in the respiratory complexes I and IV, which are correlated with an increased life span, a strong induction of the alternative oxidase, and a reduction in ROS production. However, there is no causal link between alternative oxidase level and life span. We also show that in the oxa1(ts) mutant, the extent of the defects in complexes I and IV and the life-span increase depends on the essential gene rmp1. The RMP1 protein, whose function is still unknown, can be localized in the mitochondria and/or the cytosolic compartment, depending on the developmental stage. We propose that the RMP1 protein could be involved in the process of OXA1-dependent protein insertion.

Effect of Temperature on Rates of Alternative and Cytochrome Pathway Respiration and Their Relationship with the Redox Poise of the Quinone Pool

We investigated the effect of short-term changes in temperature on alternative (Alt) and cytochrome (Cyt) pathway respiration, both in intact tissues and isolated mitochondria of 14-d-old cotyledons of soybean (Glycine max L. cv Stevens). We also established the extent to which temperature alters the interaction between the oxidizing pathways and the level of ubiquinone (UQ) reduction (UQ(r)/UQ(t)). No difference was found between the temperature coefficient of respiration (Q(10); proportional change per 10 degrees C) of Alt and Cyt pathway respiration in cotyledon slices (Q(10) = 1.92 and 1.86, respectively). In isolated mitochondria, the Q(10) of the fully activated Alt pathway (Q(10) = 2.24-2.61) was always equal to, or higher than, that of Cyt c oxidase (COX) alone (Q(10) = 2.08) and the complete Cyt pathway (Q(10) = 2.40-2.55). This was true regardless of substrate or whether ADP was present. There was little difference in the Q(10) of the Cyt pathway with or without ADP; however, the Q(10) of COX was substantially lower in the presence of an uncoupler (Q(10) = 1.61) than its absence (Q(10) = 2.08). The kinetics of Alt and Cyt pathway activity in relation to UQ(r)/UQ(t) were not affected by temperature. For a given UQ(r)/UQ(t) value, the proportion of maximum flux taking place was similar at all temperatures for both pathways (+/-ADP). However, the Q(10) of the Alt and the Cyt pathways (+ADP) increased with increasing UQ(r)/UQ(t). We conclude that the Alt pathway is not less temperature sensitive than the Cyt pathway or COX per se and that changes in the degree of control exerted by individual steps in the respiratory apparatus could result in changes in the Q(10) of mitochondrial O(2) uptake.

Effect of Temperature on Rates of Alternative and Cytochrome Pathway Respiration and Their Relationship with the Redox Poise of the Quinone Pool

We investigated the effect of short-term changes in temperature on alternative (Alt) and cytochrome (Cyt) pathway respiration, both in intact tissues and isolated mitochondria of 14-d-old cotyledons of soybean (Glycine max L. cv Stevens). We also established the extent to which temperature alters the interaction between the oxidizing pathways and the level of ubiquinone (UQ) reduction (UQr/UQt). No difference was found between the temperature coefficient of respiration (Q 10; proportional change per 10°C) of Alt and Cyt pathway respiration in cotyledon slices (Q 10 = 1.92 and 1.86, respectively). In isolated mitochondria, the Q 10 of the fully activated Alt pathway (Q 10 = 2.24–2.61) was always equal to, or higher than, that of Cytc oxidase (COX) alone (Q 10 = 2.08) and the complete Cyt pathway (Q 10 = 2.40–2.55). This was true regardless of substrate or whether ADP was present. There was little difference in the Q 10 of the Cyt pathway with or without ADP; however, theQ 10 of COX was substantially lower in the presence of an uncoupler (Q 10 = 1.61) than its absence (Q 10 = 2.08). The kinetics of Alt and Cyt pathway activity in relation to UQr/UQt were not affected by temperature. For a given UQr/UQt value, the proportion of maximum flux taking place was similar at all temperatures for both pathways (±ADP). However, the Q 10 of the Alt and the Cyt pathways (+ADP) increased with increasing UQr/UQt. We conclude that the Alt pathway is not less temperature sensitive than the Cyt pathway or COX per se and that changes in the degree of control exerted by individual steps in the respiratory apparatus could result in changes in theQ 10 of mitochondrial O2uptake.