Mitochondrial Alternative Oxidase Pathway Research Articles

Mitochondrial alternative oxidase pathway helps in nitro-oxidative stress tolerance in germinating chickpea

Mitochondrial alternative oxidase pathway helps in nitro-oxidative stress tolerance in germinating chickpea

Mitochondrial alternative oxidase pathway accelerates non-motile cell germination by enhancing respiratory carbon metabolism and maintaining redox poise in Haematococcus pluvialis

Low efficiency of the cultivation process is a major obstacle in the commercial production of Haematococcus pluvialis. Germination of red, non-motile cells is an efficient strategy for rapid acquisition of zoospores. However, the regulatory mechanisms associated with germination remain unexplored. In the present study, it was confirmed that the mitochondrial alternative oxidase (AOX) pathway accelerates H. pluvialis cell germination, and the regulatory mechanisms were clarified. When the AOX pathway was inhibited, the transcriptomic and metabonomic data revealed a downregulation in respiratory carbon metabolism and nucleotide synthesis due to NADH accumulation. This observation suggested that AOX promoted the rapid consumption of NADH, which accelerated carbohydrate and lipid catabolism, thereby producing carbon skeletons for DNA replication through respiratory metabolism. Moreover, AOX could potentially enhance germination by disturbing the abscisic acid signaling pathway. These findings provide novel insights for developing industrial cultivation models based on red-cell-germination for achieving rapid proliferation of H. pluvialis.

Photosystems under high light stress: throwing light on mechanism and adaptation

High light stress decreases the photosynthetic rate in plants due to photooxidative damage to photosynthetic apparatus, photoinhibition of PSII, and/or damage to PSI. The dissipation of excess energy by nonphotochemical quenching and degradation of the D1 protein of PSII and its repair cycle help against photooxidative damage. Light stress also activates stress-responsive nuclear genes through the accumulation of phosphonucleotide-3'-phosphoadenosine- 5'-phosphate, methylerythritol cyclodiphosphate, and reactive oxygen species which comprise the chloroplast retrograde signaling pathway. Additionally, hormones, such as abscisic acid, cytokinin, brassinosteroids, and gibberellins, play a role in acclimation to light fluctuations. Several alternate electron flow mechanisms, which offset the excess of electrons, include activation of plastid or plastoquinol terminal oxidase, cytochrome b6/f complex, cyclic electron flow through PSI, Mehler ascorbate peroxidase pathway or water-water cycle, mitochondrial alternative oxidase pathway, and photorespiration. In this review, we provided insights into high light stress-mediated damage to photosynthetic apparatus and strategies to mitigate the damage by decreasing antennae size, enhancing NPQ through the introduction of mutants, expression of algal proteins to improve photosynthetic rates and engineering ATP synthase.

Upregulation of the mitochondrial alternative oxidase pathway improves PSII function and photosynthetic electron transport in tomato seedlings under chilling stress

The aim of this study was to explore how the mitochondrial alternative oxidase (AOX) pathway alleviates photoinhibition in chilled tomato (Solanum lycopersicum) seedlings. Chilling induced photoinhibition in tomato seedlings despite the increases in thermal energy dissipation and cyclic electron flow around PSI (CEF-PSI). Chilling inhibited the function of PSII and blocked electron transport at the PSII acceptor side, however, it did not affect the oxygen-evolving complex on the donor side of PSII. Upregulation of the AOX pathway protects against photoinhibition by improving PSII function and photosynthetic electron transport in tomato seedlings under chilling stress. The AOX pathway maintained the open state of PSII and the stability of the entire photosynthetic electron transport chain. Moreover, the protective role of the AOX pathway on PSII was more important than that on PSI. However, inhibition of the AOX pathway could be compensated by increasing CEF-PSI activity under chilling stress.

Mitochondrial alternative oxidase pathway acts as an electron sink during photosynthetic induction in Rumex K-1 leaves

Chloroplast PSII photochemical efficiency is upregulated more rapidly than CO2 assimilation during photosynthesis induction, suggesting the existence of other electron sinks than that of CO2 assimilation. We hypothesized that the mitochondrial alternative oxidase (AOX) pathway could be such a sink. Inhibition of the AOX restricted light activation of the malate-oxaloacetate shuttle and caused an excessive reduction of PSI acceptor side and substantial accumulation of QA-, hindering the photosynthetic linear electron transport rate (ETR) and leading to an imbalance between light energy absorption and exploitation during photosynthetic induction. ETR limitation also restricted the formation of thylakoid pH gradient, evidenced by a decreased de-epoxidation of the xanthophyll cycle, thus preventing nonphotochemical quenching. Delayed CO2 assimilation due to thylakoid pH gradient restriction was partially reversed by exogenous ATP application. The AOX pathway acts as a photosynthetic electron sink, protecting the photosynthetic apparatus against photoinhibition and accelerating the induction of CO2 assimilation during photosynthetic induction in Rumex K-1 leaves.

Upregulation of mitochondrial alternative oxidase pathway protects photosynthetic apparatus against photodamage under chilling stress in Rumex K-1 leaves

This study aimed to clarify the role of mitochondrial alternative oxidase (AOX) pathway upregulation in photoprotection under chilling stress in Rumex K-1 leaves. Malate-oxaloacetate shuttleꞌs activity and AOX pathwayꞌs capacity were synchronously enhanced with light intensity increase both in control and chilling-treated leaves. Besides, the AOX pathway was upregulated more significantly when irradiated by light in chilling-treated leaves than that of control. These results suggested that excess reducing equivalents generated by photosynthesis were exported from chloroplasts and oxidized by the AOX pathway at a potentially higher rate in chilling-treated leaves than in control, which avoided the overaccumulation of excess reducing equivalents under light conditions. Also, inhibition of the AOX pathway led to more severe accumulation of hydrogen peroxide in chilling-treated leaves than that in control. Therefore, the mitochondrial AOX pathway was specifically upregulated and employed for sinking excess reducing equivalents produced by photosynthesis, which protected photosynthetic apparatus from photodamage under chilling stress in Rumex K-1 leaves.

Jasmonic Acid- and Ethylene-Induced Mitochondrial Alternative Oxidase Stimulates Marssonina brunnea Defense in Poplar.

Mitochondrial processes are implicated in plant response to biotic stress caused by viruses, actinomyces, bacteria and pests, but their function in defense against fungal invasion remains unclear. Here, we investigated the role and regulation of mitochondrial alternative oxidase (AOX) in response to black spot disease caused by the hemibiotrophic fungus Marssonina brunnea in poplar. M. brunnea inoculation induced the transcription of the AOX1a gene in the mitochondrial electron transport chain and of jasmonic acid (JA) and ethylene (ET) biosynthetic genes, with the accumulation of these phytohormones in poplar leaf, while inhibiting the transcript amount of the mitochondrial cytochrome c oxidase gene (COX6b) and genes related to salicylic acid (SA). Enhanced AOX reduced poplar susceptibility to M. brunnea with a higher ATP/ADP ratio while the repressed AOX caused the reverse effect. Exogenous JA and 1-aminocyclopropane-1-carboxylic acid (ACC, a biosynthetic precursor of ET) inhibited the transcript amount of COX6b and consequently increased the ratio of AOX pathway to total respiration. Furthermore, the transcription of CYS C1 and CYS D1 genes catalyzing cyanide metabolism was induced, while the cysteine (CYS) substrate levels reduced upon M. brunnea inoculation; exogenous JA and ACC mimicked the effect of M. brunnea infection on cysteine. Exogenous SA enhanced, while JA and ACC reduced, poplar susceptibility to M. brunnea. Moreover, inhibiting AOX completely prohibited JA- and ET-increased tolerance to M. brunnea in poplar. These observations indicate that the JA- and ET-induced mitochondrial AOX pathway triggers defense against M. brunnea in poplar. This effect probably involves cyanide. These findings deepen our understanding of plant-pathogenic fungi interactions.

Distinct roles of alternative oxidase pathway during the greening process of etiolated algae.

The vital function of mitochondrial alternative oxidase (AOX) pathway in optimizing photosynthesis during plant de-etiolation has been well recognized. However, whether and how AOX impacts the chloroplast biogenesis in algal cells remains unclear. In the present study, the role of AOX in regulating the reassembly of chloroplast in algal cells was investigated by treating Auxenochlorella protothecoides with salicylhydroxamic acid (SHAM), the specific inhibitor to AOX, in the heterotrophy to autotrophy transition process. Several lines of evidences including delayed chlorophyll accumulation, lagged reorganization of chloroplast structure, altered PSI/PSII stoichiometry, and declined photosynthetic activities in SHAM treated cells indicated that the impairment in AOX activity dramatically hindered the development of functioning chloroplast in algal cells. Besides, the cellular ROS levels and activities of antioxidant enzymes were increased by SHAM treatment, and the perturbation on the balance of NAD+/NADH and NADP+/NADPH ratios was also observed in A. protothecoides lacking AOX activity, indicating that AOX was essential in promoting ROS scavenging and keeping the redox homeostasis for algal chloroplast development during greening. Overall, our study revealed the essentiality of mitochondrial AOX pathway in sustaining algal photosynthetic performance and provided novel insights into the physiological roles of AOX on the biogenesis of photosynthetic organelle in algae.

Antioxidant enzymes and the mitochondrial alternative oxidase pathway play important roles in chilling tolerance of Haematococcus pluvialis at the green motile stage

During outdoor large-scale cultivation, Haematococcus pluvialis cells, especially at the green motile stage, are frequently exposed to chilling stress in winter. However, the physiological responses and adaptation mechanisms to chilling in H. pluvialis have not been characterized. In order to better understand the chilling tolerance mechanisms of H. pluvialis in the green motile stage, the responses of photosynthetic characteristics and photoprotective mechanisms to chilling were investigated. Chilling stress significantly decreased the activities of key enzymes (ribulose-1,5-bisphosphate carboxylase/oxygenase and NADP glyceraldehyde-3-phosphate dehydrogenase) in photosynthetic carbon assimilation, which would cause the accumulation of excess reducing equivalents and an imbalance of light absorption and energy utilization, leading to more severe photoinhibition. After chilling treatment, there was no induction of cyclic electron flow, non-photochemical quenching, fluorescence emission, or catalase, indicating these factors do not protect H. pluvialis from photoinhibition. However, chilling significantly enhanced the activities of antioxidant enzymes (superoxide dismutase, ascorbate peroxidase, dehydroascorbate reductase, and glutathione reductase), which efficiently dissipated excess electrons generated by photosynthetic linear electron flow (LEF). The malate-oxaloacetate shuttle was activated and the mitochondrial alternative oxidase (AOX) pathway was significantly up-regulated after chilling, indicating the transport of excess reducing equivalents generated by photosynthetic LEF to cytosol and mitochondria and oxidization by the AOX pathway, the mitochondrial non-phosphorylating pathway. As a result, the LEF was not inhibited by negative feedback and the H. pluvialis cells were well protected under chilling stress in the light, with no change in the effective quantum yield of PSII (ΦPSII) and the yield of non-regulated energy dissipation of PSII (ΦNO). The results indicate that antioxidant enzymes and the mitochondrial AOX pathway play important roles in protecting H. pluvialis at the green motile stage against chilling stress.

Mitochondrial alternative oxidase pathway protects photosynthetic apparatus against photodamage by alleviating photoinhibition in Malus hupehensis leaves under drought stress

Mitochondrial alternative oxidase pathway protects photosynthetic apparatus against photodamage by alleviating photoinhibition in <i>Malus hupehensis</i> leaves under drought stress

Aluminum directly inhibits alternative oxidase pathway and changes metabolic and redox parameters on Jatropha curcas cell culture

Aluminum (Al) toxicity has been recognized to be a main limiting factor of crop productivity in acid soil. Al interacts with cell walls disrupting the functions of the plasma membrane and is associated with oxidative damage and mitochondrial dysfunction. Jatropha curcas L. (J. curcas) is a drought resistant plant, widely distributed around the world, with great economic and medicinal importance. Here we investigated the effects of Al on J. curcas mitochondrial function and cell viability, analyzing mitochondrial respiration, phenolic compounds, reducing sugars and cell viability in cultured J. curcas cells. The results showed that at 70 μM, Al limited mitochondrial respiration by inhibiting the alternative oxidase (AOX) pathway in the respiratory chain. An increased concentration of reducing sugars and reduced concentration of intracellular phenolic compounds was observed during respiratory inhibition. After inhibition, a time-dependent upregulation of AOX mRNA was observed followed by restoration of respiratory activity and reducing sugar concentrations. Cultured J. curcas cells were very resistant to Al-induced cell death. In addition, at 70 μM, Al also appeared as an inhibitor of cell wall invertase. In conclusion, Al tolerance in cultured J. curcas cells involves a inhibition of mitochondrial AOX pathway, which seems to start an oxidative burst to induce AOX upregulation, which in turn restores consumption of O2 and substrates. These data provide new insight into the signaling cascades that modulate the Al tolerance mechanism.

Unravelling the invivo regulation and metabolic role of the alternative oxidase pathway in C3 species under photoinhibitory conditions.

The mitochondrial alternative oxidase pathway (AOP) has been suggested to act as a sink for excess reducing power generated in the chloroplast under high-light (HL) stress and thus may reduce photoinhibition. The aim of this study was to compare different species to investigate the invivo regulation and role of AOP under HL stress. The invivo activities of AOP (νalt ) and the cytochrome oxidase pathway, chlorophyll fluorescence, metabolite profiles, alternative oxidase (AOX) capacity and protein amount were determined in leaves of five C3 species under growth light and after HL treatment. Differences in respiration and metabolite levels were observed among species under growth light conditions. The HL response of νalt was highly species dependent, correlated with the AOP capacity and independent of AOX protein content. Nevertheless, significant correlations were observed between νalt , levels of key metabolites and photosynthetic parameters. The results show that the species-specific response of νalt is caused by the differential post-translational regulation of AOX. Significant correlations between respiration, metabolites and photosynthetic performance across species suggest that AOP may permit stress-related amino acid synthesis, whilst maintaining photosynthetic activity under HL stress.

Enhanced fatty acid accumulation in Isochrysis galbana by inhibition of the mitochondrial alternative oxidase pathway under nitrogen deprivation

The purpose of this study was to clarify the interrelation between the mitochondrial alternative oxidase (AOX) pathway and fatty acid accumulation in marine microalga Isochrysis galbana. Under normal conditions, the activity of the AOX pathway was maintained at a low level in I. galbana. Compared with the normal condition, nitrogen deprivation significantly increased the AOX pathway activity and fatty acid accumulation. Under nitrogen deprivation, the inhibition of the AOX pathway by salicylhydroxamic acid caused the accumulation of reducing equivalents and the over-reduction of chloroplasts in I. galbana cells, leading to a decrease in the photosynthetic O2 evolution rate. The over-production of reducing equivalents due to the inhibition of the AOX pathway under nitrogen deprivation further enhanced the accumulation of fatty acids in I. galbana cells.

Role of the mitochondrial alternative oxidase pathway in hydrogen photoproduction in Chlorella protothecoides.

The AOX pathway in C. protothecoides plays an important role in the photoprotection of PSII by alleviating the inhibition of the repair of the photodamaged PSII during H2 photoproduction. We had demonstrated that nitrogen limitation (LN) substantially enhanced H2 photoproduction in Chlorella protothecoides. In the present study, the mitochondrial alternative oxidase (AOX) pathway capacity was found to increase significantly during H2 photoproduction under LN or under LN simultaneously with sulfur deprivation (LNS) conditions. The purpose of this study was to clarify the role of the AOX pathway during H2 photoproduction in C. protothecoides. The AOX pathway can affect H2 photoproduction in the following ways: (1) consuming O2, which is favorable for the establishment of anaerobiosis; (2) consuming NADPH and competing with hydrogenase for photosynthetic electrons, which would decrease the H2 photoproduction; (3) protecting photosystem (PS) II, which is a direct electron source for H2 photoproduction, from photoinhibition. In LN and LNS cultures, the inhibition of the AOX pathway reduced the H2 photoproduction significantly, and did not increase the amount of O2. But, the inhibition of the AOX pathway decreased the maximal photochemical efficiency of PSII (F v/F m) and the actual photochemical efficiency of PSII (Φ PSII) significantly, leading to photoinhibition, which would decrease the photosynthetic electrons transferred to hydrogenase. And, the inhibition of the AOX pathway did not change the level of photoinhibition in the presence of D1 protein synthesis inhibitor chloramphenicol, indicating that the inhibition of the AOX pathway did not accelerate the photodamage to PSII directly but inhibited the repair of the photodamaged PSII. Therefore, the mitochondrial AOX pathway in C. protothecoides plays an important role in the photoprotection of PSII by alleviating the inhibition of the repair of the photodamaged PSII during H2 photoproduction, which is thus able to supply more electrons to hydrogenase under LN and LNS conditions.

Light regulation of mitochondrial alternative oxidase pathway during greening of etiolated wheat seedlings

This study deals with effects of de-etiolation (48h) of spring wheat (Triticum aestivum L., var. Irgina) seedlings on differential expression of AOX1 genes, levels of AOX protein and the alternative respiratory pathway (AP) capacity. As a result of exposure to continuous irradiation of dark-grown wheat seedlings, the respiratory activity and AP capacity in leaves significantly increased during the first 6h of studies. Expression of AOX1a was up-regulated by light and proved consistent with changes in the AP capacity. Effects on expression of AOX1c were less pronounced. Immunoblot analysis showed three distinct bands of AOX with molecular weights of 34, 36 and 38kDa, with no significant changes in the relative levels during de-etiolation. The lack of a clear correlation between AOX protein amount, AOX1a expression, and AP capacity suggests post-translational control of the enzyme activation. The AOX1a suppression and a decrease in the AP capacity correlated with the sugar pool depletion after 24h of the de-etiolation, which may mean a possible substrate dependence of the AOX activity in the green cells. More efficient malate oxidation by mitochondria as well as the higher AOX capacity during the first 6h of de-etiolation was detected, whereas respiration and AOX capacity with exogenous NADH and glycine increased after 6 and 24h, respectively. We conclude that AOX plays an important role during development of an actively photosynthesizing cell, and can rapidly adapt to changes in metabolism and photosynthesis.

Peroxisomal polyamine oxidase and NADPH-oxidase cross-talk for ROS homeostasis which affects respiration rate in Arabidopsis thaliana.

Homeostasis of reactive oxygen species (ROS) in the intracellular compartments is of critical importance as ROS have been linked with nearly all cellular processes and more importantly with diseases and aging. PAs are nitrogenous molecules with an evolutionary conserved role in the regulation of metabolic and energetic status of cells. Recent evidence also suggests that polyamines (PA) are major regulators of ROS homeostasis. In Arabidopsis the backconversion of the PAs spermidine (Spd) and spermine to putrescine and Spd, respectively, is catalyzed by two peroxisomal PA oxidases (AtPAO). However, the physiological role of this pathway remains largely elusive. Here we explore the role of peroxisomal PA backconversion and in particular that catalyzed by the highly expressed AtPAO3 in the regulation of ROS homeostasis and mitochondrial respiratory burst. Exogenous PAs exert an NADPH-oxidase dependent stimulation of oxygen consumption, with Spd exerting the strongest effect. This increase is attenuated by treatment with the NADPH-oxidase blocker diphenyleneiodonium iodide (DPI). Loss-of-function of AtPAO3 gene results to increased NADPH-oxidase-dependent production of superoxide anions ( ), but not H2O2, which activate the mitochondrial alternative oxidase pathway (AOX). On the contrary, overexpression of AtPAO3 results to an increased but balanced production of both H2O2 and . These results suggest that the ratio of /H2O2 regulates respiratory chain in mitochondria, with PA-dependent production of by NADPH-oxidase tilting the balance of electron transfer chain in favor of the AOX pathway. In addition, AtPAO3 seems to be an important component in the regulating module of ROS homeostasis, while a conserved role for PA backconversion and ROS across kingdoms is discussed.

The mitochondrial alternative oxidase pathway protects the photosynthetic apparatus against photodamage in Rumex K-1 leaves.

BackgroundIt is known that excess reducing equivalents in the form of NADPH in chloroplasts can be transported via shuttle machineries, such as the malate-oxaloacetate (OAA) shuttle, into the mitochondria, where they are efficiently oxidised by the mitochondrial alternative oxidase (AOX) respiratory pathway. Therefore, it has been speculated that the AOX pathway may protect plants from photoinhibition, but the mechanism by which this protection occurs remains to be elucidated.ResultsThe observation that the malate-OAA shuttle activity and the AOX pathway capacity increased markedly after intense light treatment in Rumex K-1 leaves indicates that excess NADPH was transported from the chloroplasts and oxidised by the AOX pathway. The inhibition of the AOX pathway by salicylhydroxamic acid (SHAM) caused the over-reduction of the photosystem I (PSI) acceptor side, as indicated by the increases in the extent of reduction of P700+. Furthermore, the photosynthetic linear electron flow was restricted, which was indicated by the decreases in the PSII electron transport rate (ETR) and the photosynthetic O2 evolution rate. The restriction of the photosynthetic linear electron flow, which generates the thylakoid ΔpH, inevitably decreased the de-epoxidation of the xanthophyll cycle (ΔPRI). Therefore, the induction of non-photochemical quenching (NPQ) was suppressed when the AOX pathway was inhibited. The effect of the inhibition of the AOX pathway on NPQ induction was less at 20 mM NaHCO3 than at 1 mM NaHCO3. The suppression of NPQ induction by the inhibition of the AOX pathway was also observed during the induction phase of photosynthesis. In addition, the inhibition of the AOX pathway increased the accumulation of hydrogen peroxide (H2O2), suggesting that the AOX pathway functions as an antioxidant mechanism.ConclusionsThe inhibition of the AOX pathway resulted in the rapid accumulation of NADPH in the chloroplasts, which caused the over-reduction of the PSI acceptor side. Furthermore, the restriction of the photosynthetic linear electron flow due to the inhibition of the AOX pathway limited the generation of the thylakoid ΔpH and suppressed the induction of NPQ. Therefore, the mitochondrial AOX pathway protected the photosynthetic apparatus against photodamage by alleviating the over-reduction of the PSI acceptor side and accelerating the induction of NPQ in Rumex K-1 leaves.

Multiple effects of inhibition of mitochondrial alternative oxidase pathway on photosynthetic apparatus in Rumex K-1 leaves

The effects of inhibition of mitochondrial alternative oxidase (AOX) respiratory pathway on photosynthetic apparatus in Rumex K-1 leaves were studied. Under high irradiance, the inhibition of AOX pathway caused over-reduction of photosystem (PS) 2 acceptor side, a decrease in the energy transfer in the PS 2 units, damage of donor side of PS 2 and decrease in pool size of electron acceptors. The inhibition of AOX pathway also decreased photosynthetic performance index (PIABS), actual photochemical efficiency (ΦPS2), photochemical quenching (qP) and photosynthetic O2 evolution rate. The results demonstrate that mitochondrial AOX pathway plays a vital role in photoprotection of photosynthetic apparatus.

Mitochondrial alternative oxidase pathway protects plants against photoinhibition by alleviating inhibition of the repair of photodamaged PSII through preventing formation of reactive oxygen species in Rumex K‐1 leaves

The purpose of this study was to explore how the mitochondrial AOX (alternative oxidase) pathway alleviates photoinhibition in Rumex K-1 leaves. Inhibition of the AOX pathway decreased the initial activity of NADP-malate dehydrogenase (EC 1.1.1.82, NADP-MDH) and the pool size of photosynthetic end electron acceptors, resulting in an over-reduction of the photosystem I (PSI) acceptor side. The over-reduction of the PSI acceptor side further inhibited electron transport from the photosystem II (PSII) reaction centers to the PSII acceptor side as indicated by an increase in V(J) (the relative variable fluorescence at J-step), causing an imbalance between photosynthetic light absorption and energy utilization per active reaction center (RC) under high light, which led to the over-excitation of the PSII reaction centers. The over-reduction of the PSI acceptor side and the over-excitation of the PSII reaction centers enhanced the accumulation of reactive oxygen species (ROS), which inhibited the repair of the photodamaged PSII. However, the inhibition of the AOX pathway did not change the level of photoinhibition under high light in the presence of the chloroplast D1 protein synthesis inhibitor chloramphenicol, indicating that the inhibition of the AOX pathway did not accelerate the photodamage to PSII directly. All these results suggest that the AOX pathway plays an important role in the protection of plants against photoinhibition by minimizing the inhibition of the repair of the photodamaged PSII through preventing the over-production of ROS.

Ethylene is involved in leafy mustard systemic resistance to Turnip mosaic virus infection through the mitochondrial alternative oxidase pathway

The role of the mitochondrial alternative oxidase (AOX) pathway and the relationship between AOX and ethylene in resistance to Turnip mosaic virus (TuMV) were investigated in systemic infected mustard leaves. Expressions of AOX1a and ethylene biosynthesis-related genes were enhanced in the upper uninoculated leaves at 6 days of postinoculation (dpi). Increased accumulation of AOX1a transcript and reduced levels of TuMV-CP were also observed after pretreatment with AA (a cytochrome pathway inhibitor) or ACC (an ethylene precursor) at non-lethal concentrations. Meanwhile, application of AVG (an ethylene biosynthesis inhibitor) mostly blocked TuMV-induced AOX activity and substantially increased TuMV susceptibility in the upper leaves. SHAM (AOX pathway inhibitor) pretreatment partially reduced the ACC-induced cyanide-resistant respiration and TuMV resistance. Conversely, AA and SHAM pretreatments had little effect on generation of ethylene. These results suggested that the TuMV-induced increase of AOX was partly involved in the ethylene pathway, and ethylene unregulated AOX in upper uninoculated mustard leaves.