Journal
BMC PLANT BIOLOGY
Volume 23, Issue 1, Pages -Publisher
BMC
DOI: 10.1186/s12870-023-04516-0
Keywords
Oxygen consumption rate; OCR; High-throughput plant mitochondria respiration assay; Plant mitochondria
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We present a novel method to measure the oxygen consumption rate in isolated mitochondria of plant seeds. The optimized protocol allows for gradual decrease in respiratory function and provides information on the activity and condition of plant mitochondria.
Background: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. Results: 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. Conclusions: 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.
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