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A Rapid and Specific Microplate Assay for the Determination of Intra- and Extracellular Ascorbate in Cultured Cells
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Mitochondrial dysfunction associated with ascorbate synthesis in plants.

Luis Miguel Mazorra Morales1, Gláucia Michelle Cosme Silva2, Diederson Bortolini Santana2

  • 1Laboratório de Melhoramento Genético Vegetal, Centro de Ciências e Tecnologias Agropecuárias, Universidade Estadual do Norte Fluminense Darcy Ribeiro (UENF), Campos dos Goytacazes, Rio de Janeiro, CEP 28013-602, Brazil; Laboratório de Ciências Físicas, Centro de Ciência Tecnologia, Universidade Estadual do Norte Fluminense Darcy Ribeiro (UENF), Campos dos Goytacazes, Rio de Janeiro, CEP 28013-602, Brazil.

Plant Physiology and Biochemistry : PPB
|June 6, 2022
PubMed
Summary

High levels of L-galactone-1,4-lactone (L-GalL) induce mitochondrial dysfunction in plants by disrupting electron transport and increasing reactive oxygen species (ROS). However, L-galactone-1,4-lactone dehydrogenase (L-GalLDH) activity aids ascorbate synthesis and plant growth under these conditions.

Keywords:
Alternative oxidaseH(2)O(2)L-galactone-1,4-lactoneL-galactone-1,4-lactone dehydrogenaseMitochondrial electron transport chainReactive oxygen species

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Isolation and Respiratory Measurements of Mitochondria from Arabidopsis thaliana
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Area of Science:

  • Plant Physiology
  • Mitochondrial Biology
  • Biochemistry

Background:

  • Mitochondria are crucial for cellular energy production but can become dysfunctional.
  • Mitochondrial dysfunction is linked to impaired energy metabolism and increased reactive oxygen species (ROS).
  • L-galactone-1,4-lactone (L-GalL), the ascorbate precursor, can induce mitochondrial dysfunction at high concentrations.

Purpose of the Study:

  • To investigate the role of L-galactone-1,4-lactone dehydrogenase (L-GalLDH) in L-GalL-induced mitochondrial dysfunction.
  • To elucidate the impact of L-GalL on mitochondrial respiration, ROS production, and alternative oxidase (AOX) activity.
  • To assess the overall effect of high L-GalL concentrations on plant growth and performance.

Main Methods:

  • Induction of mitochondrial dysfunction using high L-GalL concentrations in plant mitochondria.
  • Utilized RNAi-plant lines with silenced L-GalLDH activity to determine enzyme dependency.
  • Employed respiratory inhibitors and proteomic analysis to study electron transport, ROS formation, and protein alterations.
  • Compared growth performance between wild-type and L-GalLDH-RNAi plants under high L-GalL conditions.

Main Results:

  • High L-GalL levels caused mitochondrial dysfunction, impaired electron partitioning, and ROS over-production, dependent on L-GalLDH activity.
  • Alternative respiration capacity, particularly alternative oxidase (AOX) activity, was reduced by L-GalL, but supported ascorbate synthesis.
  • ROS formation and dysfunction correlated more with reduced cytochrome oxidase (COX) and AOX activities than with complexes I and III.
  • Proteomic analysis revealed changes in proteins related to oxidative stress and energy metabolism.
  • Supra-optimal L-GalL concentrations were not detrimental; L-GalLDH activity positively influenced growth in wild-type plants compared to RNAi lines.

Conclusions:

  • L-GalL-induced mitochondrial dysfunction is mediated by L-GalLDH activity and affects key respiratory enzymes like AOX.
  • Despite causing dysfunction, L-GalLDH and alternative respiration play a role in supporting ascorbate synthesis and plant adaptation.
  • L-GalLDH activity can be beneficial for plant growth under high L-GalL conditions, suggesting a complex regulatory role.