Abstract
Reactive oxygen species (ROS) are produced throughout plant cells as a by-product of electron transfer processes. While highly oxidative and potentially damaging to a range of biomolecules, there exists a suite of ROS-scavenging antioxidant strategies that maintain a redox equilibrium. This balance can be disrupted in the event of cellular stress leading to increased ROS levels, which can act as a useful stress signal but, in excess, can result in cell damage and death. As crop plants become exposed to greater degrees of multiple stresses due to climate change, efforts are ongoing to engineer plants with greater stress tolerance. It is therefore important to understand the pathways underpinning ROS-mediated signalling and damage, both through measuring ROS themselves and other indicators of redox imbalance. The highly reactive and transient nature of ROS makes this challenging to achieve, particularly in a way that is specific to individual ROS species. In this review, we describe the range of chemical and biological tools and techniques currently available for ROS and redox marker measurement in plant cells and tissues. We discuss the limitations inherent in current methodology and opportunities for advancement.
Highlights
The evolution of aerobic life 2.2 billion years ago corresponds with the ability of cells to exploit the availability of oxygen and its electron accepting ability to generate energy through oxidative phosphorylation
NADH oxidation and oxygen reduction to water maintain a proton gradient across the mitochondrial membrane, which is the driving force for ATP synthesis. The efficiency of this process relies on tight coupling of electron transport in the mitochondrial membrane and sufficient oxygen availability to accept the electrons generated by oxidation of NADH in the electron transport chain
We provide an overview of the state-of-the-art with respect to the methods and tools used to detect and quantify both Reactive oxygen species (ROS) directly and other fluctuating redox markers in plant cells, including the resultant oxidative modifications
Summary
Superoxide can be converted into other ROS (Fig. 1A) and the impact of the altered redox status in a cell or subcellular region depends very much on the nature, location, intensity and duration of the species formed. There are multiple mechanisms in place to ‘mop up’ excessive ROS species in the form of both small molecule buffers (e.g. glutathione, ascorbate) and enzymes (e.g. superoxide dismutase, catalase). This balance of ROS formation and ROS scavengers enables cells to survive inevitable ROS production without excessive damage. This balance can become disrupted in certain, usually stressful, conditions In plants, such conditions include pathogen attack, heat, drought and flooding.[1] An increase in ROS levels beyond the ability of the cell to buffer the oxidative activity will increase the oxidative status of the cell. Remaining challenges in effective measurement of ROS and redox markers in plants
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