High-throughput method for Oxygen Consumption Rate measurement (OCR) in plant mitochondria

Conventional methods to measure oxygen consumption, such as Clark-type electrodes, have limitations such as requiring a large amount of starting material. Moreover, commercially available kits for high-throughput methods are usually optimized for animal cells and mitochondria. Here, we present a nov...

Full description

Saved in:
Bibliographic Details
Published in:BMC plant biology 2023-10, Vol.23 (1), p.1-496, Article 496
Main Authors: Fuchs, Hanna, Malecka, Arleta, Budzinska, Adrianna, Jarmuszkiewicz, Wieslawa, Ciszewska, Liliana, Staszak, Aleksandra M, Kijowska-Oberc, Joanna, Ratajczak, Ewelina
Format: Article
Language:English
Subjects:
Acids
Aging
Analysis
Botanical research
Cells
Electron transport
High-throughput plant mitochondria respiration assay
High-throughput screening (Biochemical assaying)
Measurement
Metabolism
Methods
Mitochondria
OCR
Oxidative phosphorylation
Oxygen
Oxygen consumption
Oxygen consumption rate
Phosphorylation
Physiological aspects
Plant mitochondria
Proteins
Reagents
Respiration
Respiratory function
Seeds
Sucrose
Citations: Items that this one cites
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Conventional methods to measure oxygen consumption, such as Clark-type electrodes, have limitations such as requiring a large amount of starting material. Moreover, commercially available kits for high-throughput methods are usually optimized for animal cells and mitochondria. Here, we present a novel method to measure the oxygen consumption rate using a high-throughput assay in isolated mitochondria of European beech seeds. To perform the measurements, we adapted the Agilent Seahorse XF Cell Mito Stress Test Kit protocol for measurements on plant mitochondria. The optimized protocol for OCR measurement of mitochondria isolated from beech seeds allowed the observation of storage period-dependent gradual decreases in non-phosphorylating respiration, phosphorylating respiration and maximal FCCP-stimulated respiration. The longer the seeds were stored, the greater the impairment of respiratory function. Thanks to this method it is possible to minimize the amount of plant material and conduct research to obtain information on the respiratory condition and activity of plant mitochondria, including the efficiency of oxidative phosphorylation and the maximum oxidative capacity of the respiratory chain. We demonstrated that the improved protocol is suitable for study of plant material.
ISSN:1471-2229
1471-2229
DOI:10.1186/s12870-023-04516-0