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Arabidopsis plants with altered levels of alternative oxidase

ABSTRACT: In higher plants, various developmental and environmental conditions enhance expression of the mitochondrial alternative oxidase (AOX). In this work transgenic Arabidopsis thaliana plants were generated that either overexpress AOX or inhibit its synthesis. Gene expression following antimycin A treatment, which inhibits the cytochrome pathway in mitochondria, was studied in an AOX overexpressor line. The role of AOX in regulation of reactive oxygen species (ROS) in leaves was studied in the transgenic lines. The transgenic lines were also used to investigate the mitochondria-chloroplasts interaction in assays performed at three-times growth light. For most of the parameters measured AOX antisense lines and WT plants showed a very similar response whereas AOX sense lines differed in several aspects. An unexpected result was that AOX overexpressors had higher cellular ATP/ADP ratios as compared to WT and antisense lines. This was linked to decreased photosynthetic CO2-assimilation and O2-evolution, closed PSII reaction centers and altered Calvin cycle metabolite contents. We conclude that increased mitochondrial AOX capacity can lead to a drain of electrons from the chloroplasts causing an unfavorable redox status and thereby inhibit photosynthesis.

ORGANISM(S): Arabidopsis thaliana

SUBMITTER: Jianping Yu

PROVIDER: E-GEOD-4113 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Arabidopsis plants with altered levels of alternative oxidase

Project description:In higher plants, various developmental and environmental conditions enhance expression of the mitochondrial alternative oxidase (AOX). In this work transgenic Arabidopsis thaliana plants were generated that either overexpress AOX or inhibit its synthesis. Gene expression following antimycin A treatment, which inhibits the cytochrome pathway in mitochondria, was studied in an AOX overexpressor line. The role of AOX in regulation of reactive oxygen species (ROS) in leaves was studied in the transgenic lines. The transgenic lines were also used to investigate the mitochondria-chloroplasts interaction in assays performed at three-times growth light. For most of the parameters measured AOX antisense lines and WT plants showed a very similar response whereas AOX sense lines differed in several aspects. An unexpected result was that AOX overexpressors had higher cellular ATP/ADP ratios as compared to WT and antisense lines. This was linked to decreased photosynthetic CO2-assimilation and O2-evolution, closed PSII reaction centers and altered Calvin cycle metabolite contents. We conclude that increased mitochondrial AOX capacity can lead to a drain of electrons from the chloroplasts causing an unfavorable redox status and thereby inhibit photosynthesis. Keywords: Genetic modification

2006-01-31 | GSE4113 | GEO

Response to antimycin A in wild type Arabidopsis and in alternative oxidase overexpresser

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Response to antimycin A in wild type Arabidopsis and in alternative oxidase overexpresser

Project description:In higher plants, various developmental and environmental conditions enhance expression of the mitochondrial alternative oxidase (AOX). In this work transgenic Arabidopsis thaliana plants were generated that either overexpress AOX or inhibit its synthesis. Gene expression following antimycin A treatment, which inhibits the cytochrome pathway in mitochondria, was studied in an AOX overexpressor line. An accumulation of AOX protein blocked normal responses of a number of genes, including some AOX genes. This result indicates that AOX can generate feedback to inhibit induction of its own synthesis. Two wild type replicates: Poly-A RNA from two batches of plants were labeled with dye-swaps then hybridized to custom-designed arrays containing two sets of spotted DNA elements. Two S5 replicates: Poly-A RNA from two batches of plants were labeled with dye-swaps then hybridized to custom-designed arrays containing three sets of spotted DNA elements.

2010-07-01 | E-GEOD-4141 | biostudies-arrayexpress

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Project description:Transcriptional profiling of Arabidopsis thaliana 12-days old seedlings comparing Col-0 wild type with transgenic plants with altered expression of dual-targetting plastid/mitochondrial organellar RNA-polymerase RPOTmp. Transgenic plants used for experiment were: overexpressor plants obtained by transformation of Col-0 WT plants with genetic constructs created in [Tarasenko et al., 2016] contained catalytic part of RPOTmp enzyme with transit peptides of RPOTm (mitochondrial) and RPOTp (plastid) by agrobacterial transformation; plants with complementation of RPOTmp functions in mitochondria or chloroplasts obtained from transformation of GABI_286E07 rpotmp knockout-mutant plants with genetic constructs created in [Tarasenko et al., 2016]. Goal was to determine the effects of RPOTmp knockout/overexpression on global Arabidopsis thaliana gene expression.

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Expression of Ciona intestinalis alternative oxidase (AOX) in a mouse model of cardiac ischemia-reperfusion

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Transcriptome analysis of the shoots of Arabidopsis wild-type and mutants deficient in either or both activities of nitrate reductase and alternative oxidase

Project description:The conversion of nitrate to ammonium, known as nitrate reduction, consumes large amounts of reductants in plants. Previous studies have observed that mitochondrial alternative oxidase (AOX) is upregulated under conditions of limited nitrate reduction, such as low or no nitrate availability, or when ammonium serves as the sole nitrogen (N) source. Electron transfer from ubiquinone to AOX bypasses the proton-pumping complexes III and IV, thereby consuming reductants efficiently. Therefore, the upregulation of AOX under conditions of limited nitrate reduction may help dissipate excessive reductants and mitigate oxidative stress. However, firm evidence supporting this hypothesis is lacking due to the absence of experimental systems capable of directly analyzing the relationship between nitrate reduction and AOX. To address this gap, we developed a novel culturing system that allows for the manipulation of nitrate reduction and AOX activities separately, without inducing N starvation, ammonium toxicity, or disrupting the nitrate signal. Using this system, we investigated genome-wide gene expression with RNA-seq to gain insight into the relationship between AOX and nitrate reduction.

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