Functional analysis reveals effects of tobacco alternative oxidase gene (NtAOX1a) on regulation of defence responses against abiotic and biotic stresses

2009 ◽  
Vol 29 (6) ◽  
pp. 375-383 ◽  
Author(s):  
Yi Zhang ◽  
Dongmei Xi ◽  
Jian Wang ◽  
Dongfang Zhu ◽  
Xingqi Guo
Keyword(s):  
Oxidative Stress ◽  
Low Temperature ◽  
Alternative Oxidase ◽  
Alternative Pathway ◽  
Respiratory Pathway ◽  
Oxidase Gene ◽  
Biotic Stresses ◽  
Alternative Respiratory Pathway ◽  
Cytochrome Pathway ◽  
Defence Responses

Mitochondrial AOX (alternative oxidase) is the terminal oxidase of the CN (cyanide)-resistant alternative respiratory pathway in plants. To investigate the role of the tobacco AOX gene (NtAOX1a) (where Nt is Nicotiana tabacum) under deleterious conditions which could induce ROS (reactive oxygen species) accumulation, we generated and characterized a number of independent transgenic tobacco (N. tabacum) lines with altered NtAOX1a gene expression and AP (alternative pathway) capacity. AOX efficiently inhibited the production of low-temperature-induced H2O2 and might be a major enzyme for scavenging H2O2 at low temperature. Furthermore, NtAOX1a may act as a regulator of KCN-induced resistance to TMV (tobacco mosaic virus) through the regulation of H2O2. Notably, a moderate accumulation of H2O2 under the control of NtAOX1a was crucial in viral resistance. Analysis of seed germination indicated an important role for NtAOX1a in germination under H2O2-induced oxidative stress when the CP (cytochrome pathway) was inhibited. These results demonstrate that NtAOX1a is necessary for plants to survive low temperature, pathogen attack and oxidative stress by scavenging ROS under these adverse conditions when the CP is restricted.

Cloning and Analysis of the Alternative Oxidase Gene of Neurospora crassa

Genetics ◽  
1996 ◽  
Vol 142 (1) ◽  
pp. 129-140 ◽  
Author(s):  
Qiuhong Li ◽  
R Gary Ritzel ◽  
Lesley L T McLean ◽  
Lee McIntosh ◽  
Tak Ko ◽  
...  
Keyword(s):  
Neurospora Crassa ◽  
Alternative Oxidase ◽  
Electron Flow ◽  
Mitochondrial Protein ◽  
Transcriptional Level ◽  
Wild Type ◽  
Respiratory Pathway ◽  
Oxidase Gene ◽  
Alternative Respiratory Pathway ◽  
Cytochrome Pathway

Mitochondria of Neurospora crassa contain a cyanide-resistant alternative respiratory pathway in addition to the cytochrome pathway. The alternative oxidase is present only when electron flow through the cytochrome chain is restricted. Both genomic and cDNA copies for the alternative oxidase gene have been isolated and analyzed. The sequence of the predicted protein is homologous to that of other species. The mRNA for the alternative oxidase is scarce in wild-type cultures grown under normal conditions, but it is abundant in cultures grown in the presence of chloramphenicol, an inhibitor of mitochondrial protein synthesis, or in mutants deficient in mitochondrial cytochromes. Thus, induction of alternative oxidase appears to be at the transcriptional level. Restriction fragment length polymorphism mapping of the isolated gene demonstrated that it is located in a position corresponding to the aod-1 locus. Sequence analysis of mutant aod-1 alleles reveals mutations affecting the coding sequence of the alternative oxidase. The level of aod-1 mRNA in an aod-2 mutant strain that had been grown in the presence of chloramphenicol was reduced several fold relative to wild-type, supporting the hypothesis that the product of aod-2 is required for optimal expression of aod-1.

scholarly journals Temperature-dependent alterations of respiration in leaves of two selected vascular plant species: the role of the alternative oxidase pathway

Bragantia ◽  
2002 ◽  
Vol 61 (2) ◽  
pp. 111-114 ◽  
Author(s):  
Natalia V. Pystina ◽  
Roman A. Danilov
Keyword(s):  
Alternative Oxidase ◽  
Alternative Pathway ◽  
Vascular Plant ◽  
Relative Activity ◽  
Temperature Dependent ◽  
Respiratory Pathway ◽  
Alternative Respiratory Pathway ◽  
Benzhydroxamic Acid ◽  
Increasing Temperature

Effects of higher temperatures on respiration and activity of alternative oxidase (AOX) were studied in mature leaves of Ajuga reptans L. and Rhodiola rosea L. Total respiration in both A. reptans and R. rosea increased exponentially with the increasing temperature of 10 °C to 35 °C. Respiration in the presence of benzhydroxamic acid (BHAM) also increased exponentially in accordance with the increasing temperature in the leaves of both A. reptans and R. rosea. Relative activity of the alternative pathway decreased significantly in the leaves of A. reptans with increasing temperatures. However, an increase in the relative activity of the alternative pathway was detected in the leaves of R. rosea. Thermoresistance of the alternative respiratory pathway was considered to be higher in R. rosea compared to A. reptans. We suppose that A. reptans and R. rosea have different mechanisms regulating partitioning of electrons to the alternative respiratory pathway.

scholarly journals Effect of growth at low temperature on the alternative pathway respiration in Acanthamoeba castellanii mitochondria.

2001 ◽  
Vol 48 (3) ◽  
pp. 729-737 ◽  
Author(s):  
W Jarmuszkiewicz ◽  
O Fraczyk ◽  
L Hryniewiecka
Keyword(s):  
Low Temperature ◽  
Alternative Oxidase ◽  
Alternative Pathway ◽  
Plant Mitochondria ◽  
Physiological Role ◽  
Acanthamoeba Castellanii ◽  
Control Culture ◽  
Cytochrome Pathway ◽  
Coupling Parameters ◽  
Unicellular Organisms

Mitochondria of amoeba Acanthamoeba castellanii in addition to the conventional cytochrome pathway possess, like plant mitochondria, a cyanide-resistant alternative quinol oxidase. In mitochondria isolated from amoeba batch culture grown temporarily at low temperature (6 degrees C), higher respiration was accompanied by lower coupling parameters as compared to control culture (grown at 28 degrees C). In the presence of benzohydroxamate, respiratory rates and coupling parameters were similar in both types of mitochondria indicating that growth in cold conditions did not disturb the cytochrome pathway. Increased contribution of alternative oxidase in total mitochondrial respiration in low-temperature-grown amoeba cells was confirmed by calculation of its contribution using ADP/O measurements. Furthermore, in mitochondria from low-temperature- grown cells the content of the alternative oxidase was increased and correlated with the increase in the unstimulated and GMP-stimulated cyanide-resistant respiratory activity. A possible physiological role of higher activity of alternative oxidase as response to growth at a low temperature in unicellular organisms, such as amoeba, is discussed.

The cytochrome oxidase inhibitor azide also inhibits the alternative pathway of Neurospora crassa

1984 ◽  
Vol 62 (2-3) ◽  
pp. 129-136 ◽  
Author(s):  
Peter J. Bridge ◽  
Helmut Bertrand
Keyword(s):  
Neurospora Crassa ◽  
Cytochrome Oxidase ◽  
Alternative Oxidase ◽  
Alternative Pathway ◽  
Oxidase Inhibitor ◽  
The Novel ◽  
Respiratory Pathway ◽  
Maximal Inhibition ◽  
Cytochrome Pathway ◽  
Mutant Strains

Mutant strains of Neurospora crassa were used to investigate the effects of the inhibitor azide upon respiration mediated by either the cyanide-insensitive alternative mitochondrial respiratory pathway or the "standard" cyanide-sensitive cytochrome pathway. The use of appropriate cytochrome-oxidase-deficient and alternative-oxidase-deficient mutants permitted respiration to be restricted to the pathway of choice. In addition to its known effect upon cytochrome oxidase, azide was found to inhibit the alternative pathway of Neurospora with half-maximal inhibition occurring at 1.3 mM azide. This finding indicates that the activities of at least some of the "novel" azide-sensitive oxidases that have been discovered recently in organisms with inducible or constitutive cyanide-insensitive respiratory systems might be attributable to the "standard" salicylhydroxamate-sensitive mitochondrial alternative pathway.

Arabidopsis mtHSC70-1 plays important roles in the establishment of COX-dependent respiration and redox homeostasis

2019 ◽  
Vol 70 (20) ◽  
pp. 5575-5590 ◽  
Author(s):  
Shan-Shan Wei ◽  
Wei-Tao Niu ◽  
Xiao-Ting Zhai ◽  
Wei-Qian Liang ◽  
Meng Xu ◽  
...  
Keyword(s):  
Alternative Oxidase ◽  
Redox Homeostasis ◽  
Superoxide Dismutase 1 ◽  
Respiratory Pathway ◽  
Cellular Processes ◽  
Growth Defects ◽  
Alternative Respiratory Pathway ◽  
Abnormal Mitochondria ◽  
Shock Proteins ◽  
Severe Growth

Abstract The 70 kDa heat shock proteins function as molecular chaperones and are involved in diverse cellular processes. However, the functions of the plant mitochondrial HSP70s (mtHSC70s) remain unclear. Severe growth defects were observed in the Arabidopsis thaliana mtHSC70-1 knockout lines, mthsc70-1a and mthsc70-1b. Conversely, the introduction of the mtHSC70-1 gene into the mthsc70-1a background fully reversed the phenotypes, indicating that mtHSC70-1 is essential for plant growth. The loss of mtHSC70-1 functions resulted in abnormal mitochondria and alterations to respiration because of an inhibition of the cytochrome c oxidase (COX) pathway and the activation of the alternative respiratory pathway. Defects in COX assembly were observed in the mtHSC70-1 knockout lines, leading to decreased COX activity. The mtHSC70-1 knockout plants have increased levels of reactive oxygen species (ROS). The introduction of the Mn-superoxide dismutase 1 (MSD1) or the catalase 1 (CAT1) gene into the mthsc70-1a plants decreased ROS levels, reduced the expression of alternative oxidase, and partially rescued growth. Taken together, our data suggest that mtHSC70-1 plays important roles in the establishment of COX-dependent respiration.

scholarly journals Role of Alternative Oxidase Gene in Pathogenesis of Cryptococcus neoformans

2003 ◽  
Vol 71 (10) ◽  
pp. 5794-5802 ◽  
Author(s):  
Shamima Akhter ◽  
Henry C. McDade ◽  
Jenifer M. Gorlach ◽  
Garrett Heinrich ◽  
Gary M. Cox ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Cryptococcus Neoformans ◽  
Mutant Strain ◽  
Alternative Oxidase ◽  
Temperature Sensitive ◽  
Wild Type ◽  
Phagocytic Cells ◽  
Pathogenic Yeast ◽  
Oxidase Gene ◽  
Oxidative Pathway

ABSTRACT We identified a homologue of the alternative oxidase gene in a screen to identify genes that are preferentially transcribed in response to a shift to 37°C in the human-pathogenic yeast Cryptococcus neoformans. Alternative oxidases are nucleus-encoded mitochondrial proteins that have two putative roles: they can function in parallel with the classic cytochrome oxidative pathway to produce ATP, and they may counter oxidative stress within the mitochondria. The C. neoformans alternative oxidase gene (AOX1) was found to exist as a single copy in the genome, and it encodes a putative protein of 401 amino acids. An aox1 mutant strain was created using targeted gene disruption, and the mutant strain was reconstituted to wild type using a full-length AOX1. Compared to both the wild-type and reconstituted strains, the aox1 mutant strain was not temperature sensitive but did have significant impairment of both respiration and growth when treated with inhibitors of the classic cytochrome oxidative pathway. The aox1 mutant strain was also found to be more sensitive to the oxidative stressor tert-butyl hydroperoxide. The aox1 mutant strain was significantly less virulent than both the wild type and the reconstituted strain in the murine inhalational model, and it also had significantly impaired growth within a macrophage-like cell line. These data demonstrate that the alternative oxidase of C. neoformans can make a significant contribution to metabolism, has a role in the yeast's defense against exogenous oxidative stress, and contributes to the virulence composite of this organism, possibly by improving survival within phagocytic cells.

scholarly journals Kinetic analysis of the mitochondrial quinol-oxidizing enzymes during development of thermogenesis in Arum maculatum L

1996 ◽  
Vol 317 (1) ◽  
pp. 313-319 ◽  
Author(s):  
Graeme R. LEACH ◽  
Klaas KRAB ◽  
David G. WHITEHOUSE ◽  
Anthony L. MOORE
Keyword(s):  
Respiratory Rate ◽  
Succinate Dehydrogenase ◽  
Oxidase Activity ◽  
Alternative Oxidase ◽  
Alternative Pathway ◽  
Respiratory Control ◽  
Stage Of Development ◽  
Activation Level ◽  
Cytochrome Pathway ◽  
Arum Maculatum

The dependence of the rate of oxygen uptake upon the ubiquinone (Q)-pool reduction level in mitochondria isolated during the development of thermogenesis of Arum maculatum spadices has been investigated. At the α-stage of development, the respiratory rate was linearly dependent upon the reduction level of the Q-pool (Qr) both under state-3 and -4 conditions. Progression through the β/γ to the Δ-stage resulted in a non-linear dependence of the state-4 rate on Qr. In the Δ-stage of development, both state-3 and -4 respiratory rates were linearly dependent upon Qr due to a shift in the engagement of the alternative oxidase to lower levels of Qr. Western blot analysis revealed that increased alternative oxidase activity could be correlated with expression of a 35 kDa protein. Respiratory control was only observed with mitochondria in the α-stage of development. At the β/γ-stage of development, the addition of ADP resulted in a significant oxidation of the Q-pool which was accompanied by a decrease in the respiratory rate. This was due either to decreased contribution of the alternative pathway to the overall respiratory rate under state 3 or by deactivation of succinate dehydrogenase activity by ADP. Cold-storage of the spadices at the β-stage of development led to increased activity of both the cytochrome pathway and succinate dehydrogenase, without any change in alternative oxidase activity. Results are discussed in terms of how changes in the activation level of the alternative oxidase and succinate dehydrogenase influence the activity and engagement of the quinol-oxidizing pathways during the development of thermogenesis in A. maculatum.

scholarly journals An alternative respiratory pathway on Candida krusei: implications on susceptibility profile and oxidative stress

2012 ◽  
Vol 12 (4) ◽  
pp. 423-429 ◽  
Author(s):  
Sofia Costa-de-Oliveira ◽  
Belém Sampaio-Marques ◽  
Matilde Barbosa ◽  
Elisabete Ricardo ◽  
Cidália Pina-Vaz ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Candida Krusei ◽  
Respiratory Pathway ◽  
Alternative Respiratory Pathway ◽  
And Oxidative Stress
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