Methods and approaches to study plant mitochondrial alternative oxidase

The alternative oxidase is a non‐proton motive ‘alternative’ to electron transport through the cytochrome pathway. Despite its wasteful nature in terms of energy conservation, the pathway is likely present throughout the plant kingdom and appears to be expressed in most plant tissues. A small alternative oxidase gene family exists, the members of which are differentially expressed in response to environmental, developmental and other cell signals. The alternative oxidase enzyme possesses tight biochemical regulatory properties that determine its ability to compete with the cytochrome pathway for electrons. Studies show that alternative oxidase can be a prominent component of total respiration in important crop species. All these characteristics suggest this pathway plays an important role in metabolism and/or other aspects of cell physiology. This brief review is an introduction to experimental methods and approaches applicable to different areas of alternative oxidase research. We hope it provides a framework for further investigation of this fascinating component of primary plant metabolism.