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Title:           Activation of alternative oxidase ensures carbon supply for ethylene and carotenoid biosynthesis during tomato fruit ripening
Subject:         DOI: 10.1093/plphys/kiaf516; Plant Physiology, 199, 3, 2025-10-16.; Abstract: Tomato (Solanum lycopersicum) is a climacteric fruit displaying a respiration peak at the onset of ripening accompanied by increased synthesis of ethylene and carotenoid pigments. Chromoplast and mitochondrial respiration participate at different stages of fruit ripening, but their in vivo regulation and function remain unclear. We determined the in vivo activities of the mitochondrial alternative oxidase (AOX) and cytochrome oxidase pathways and quantified the levels of respiratory- and ripening-related gene transcripts, primary metabolites and carotenoids in ripening tomato fruits with or without functional chromorespiration. Furthermore, we carried out physiological, molecular, and metabolic analyses of CRISPR–Cas9 mutants defective in AOX1a, the main AOX isoform upregulated during tomato fruit ripening. We confirmed that plastid terminal oxidase-dependent chromorespiration is only relevant at late stages of ripening and found that in vivo AOX activity significantly increased at the breaker stage, becoming the main contributor to climacteric respiration when ripening initiates. This activation did not correlate with gene expression but was likely due to increased levels of AOX activators such as pyruvate (a metabolic precursor of carotenoids), 2-oxoglutarate, and succinate. A strong alteration of ripening-related metabolites was observed in aox1a mutant fruits, highlighting a key role of the AOX pathway at the onset of ripening. Our data suggest that increased supply of tricarboxylic acid cycle intermediates at the climacteric stage allosterically enhances AOX activity, thus allowing the reoxidation of NAD(P)H to ensure carbon supply for triggering ethylene and carotenoid biosynthesis.
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Author:          Ariadna Iglesias-Sanchez
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      Title: Activation of alternative oxidase ensures carbon supply for ethylene and carotenoid biosynthesis during tomato fruit ripening
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       Item: 
        Title: Introduction
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       Item: 
        Title: Results
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        Children: 
         Item: 
          Title: Measurements of oxygen consumption and isotope discrimination during tomato fruit ripening
          Destination: seckiaf516-s2.1
         Item: 
          Title: Transcript and metabolite changes in WT and ghost tomato fruits during ripening
          Destination: seckiaf516-s2.2
         Item: 
          Title: Generation and fruit development of AOX1a knock-out lines
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         Item: 
          Title: Respiration and ripening-related gene expression in aox1a mutant fruits
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         Item: 
          Title: Primary and carotenoid metabolism in WT and aox1a mutant fruits
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         Item: 
          Title: In vivo regulation of fruit respiration and consequences for energy metabolism during tomato fruit ripening
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         Item: 
          Title: Coordinated changes in primary metabolism and electron partitioning to AOX allows for the accumulation of key ripening-related metabolic precursors
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          Title: Plant material and growth conditions
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         Item: 
          Title: Respiration and oxygen isotope discrimination analysis
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          Title: Gene expression analysis
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          Title: Western blot analysis
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          Title: Primary metabolite and carotenoid profiling
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    Title: Activation of alternative oxidase ensures carbon supply for ethylene and carotenoid biosynthesis during tomato fruit ripening
    Author: Ariadna Iglesias-Sanchez
    Subject: DOI: 10.1093/plphys/kiaf516; Plant Physiology, 199, 3, 2025-10-16.; Abstract: Tomato (Solanum lycopersicum) is a climacteric fruit displaying a respiration peak at the onset of ripening accompanied by increased synthesis of ethylene and carotenoid pigments. Chromoplast and mitochondrial respiration participate at different stages of fruit ripening, but their in vivo regulation and function remain unclear. We determined the in vivo activities of the mitochondrial alternative oxidase (AOX) and cytochrome oxidase pathways and quantified the levels of respiratory- and ripening-related gene transcripts, primary metabolites and carotenoids in ripening tomato fruits with or without functional chromorespiration. Furthermore, we carried out physiological, molecular, and metabolic analyses of CRISPR Cas9 mutants defective in AOX1a, the main AOX isoform upregulated during tomato fruit ripening. We confirmed that plastid terminal oxidase-dependent chromorespiration is only relevant at late stages of ripening and found that in vivo AOX activity significantly increased at the breaker stage, becoming the main contributor to climacteric respiration when ripening initiates. This activation did not correlate with gene expression but was likely due to increased levels of AOX activators such as pyruvate (a metabolic precursor of carotenoids), 2-oxoglutarate, and succinate. A strong alteration of ripening-related metabolites was observed in aox1a mutant fruits, highlighting a key role of the AOX pathway at the onset of ripening. Our data suggest that increased supply of tricarboxylic acid cycle intermediates at the climacteric stage allosterically enhances AOX activity, thus allowing the reoxidation of NAD(P)H to ensure carbon supply for triggering ethylene and carotenoid biosynthesis.
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  Checksum: f12125e8
   Type: CRC32
  Checksum: 60e67e9468a9157170b1984b3777917c
   Type: MD5
  Checksum: c6a8f7a92fc425d21f5e6f05ff47739593e2a5d8
   Type: SHA-1
  Checksum: 274d6ee03bf44274f7f7b95fa4180eb228f24d8e55ba014557a4150d6d6d5048
   Type: SHA-256