Dark respiration measurement from arabidopsis shoots

Jose P. Fonseca, Marcus Griffiths, Larry M. York, Kirankumar S. Mysore

Research output: Contribution to journalArticlepeer-review

3 Scopus citations

Abstract

Dark respiration refers to experimental measures of leaf respiration in the absence of light, done to distinguish it from the photorespiration that occurs during photosynthesis. Dark aerobic respiration reactions occur solely in the mitochondria and convert glucose molecules from cytoplasmatic glycolysis and oxygen into carbon dioxide and water, with the generation of ATP molecules. Previous methods typically use oxygen sensors to measure oxygen depletion or complicated and expensive photosynthesis instruments to measure CO2 accumulation. Here, we provide a detailed, step-by-step approach to measure dark respiration in plants by recording CO2 fluxes of Arabidopsis shoot and root tissues. Briefly, plants are dark acclimated for 1 hour, leaves and roots are excised and placed separately in airtight chambers, and CO2 accumulation is measured over time with standard infrared gas analyzers. The time-series data is processed with R scripts to produce dark respiration rates, which can be standardized by fresh or dry tissue mass. The current method requires inexpensive infrared gas analyzers, off-the-shelf parts for chambers, and publicly available data analysis scripts.

Original languageEnglish
Article numbere4181
JournalBio-protocol
Volume11
Issue number19
DOIs
StatePublished - Oct 5 2021
Externally publishedYes

Funding

This work was supported by the Noble Research Institute, LLC. This protocol was derived from Fonseca et al. (2020) (doi: 10.1104/pp.20.00950).

Keywords

  • CO2
  • Dark respiration
  • Flux
  • Mitochondria
  • NFS1

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