Analyses of reactive oxygen species and antioxidants in relation to seed longevity and germination

Evidence is emerging that reactive oxygen species (ROS) and antioxidants, together with plant hormones and other reactive species, such as reactive nitrogen species, are part of signalling networks pertinent to plant stress responses, cell division, and cell death. Consequently, they play pivotal roles in the regulation of seed development and maturation, germination and dormancy, seedling establishment, and seed ageing. Importantly, ROS, although essentially required at low concentrations, must be kept under stringent control by antioxidants. If the balance between pro- and antioxidative processes is lost and ROS production prevails, oxidative stress is the result, which can induce cell death and ultimately seed death. This chapter offers a variety of protocols for the determination of ROS, antioxidants, and stress markers aimed at enabling the reader to quantify these compounds. Protocols are also described to visualize ROS and localize the sites of ROS production, hoping to stimulate more research into ROS signalling and antioxidant control in key physiological and biochemical processes in seeds.