Changes of paramagnetic species in cereal grains upon short-term ozone action as a marker of oxidative stress tolerance

Abstract

The increase of the concentration of ozone in the atmosphere, being the direct source of reactive oxygen species, results in the yield loss of agronomic crops. On the other hand, ozone is also used as a protector against microorganisms, living in plants and present in materials obtained from them, dangerous for human and animal health. In this work it has been studied if ozone in doses similar to those used for removal of microorganisms can have significant influence on the generation of stable organic radicals and changes in the character of transition metal ions and in the antioxidative biochemical parameters of cereal grains. The aim of this work was to find if the response of grains of three cereals (wheat, oat and barley) to ozone depended on their oxidative stress tolerance. The influence of direct short-term ozone application on grains of these cereals, each represented by two genotypes with different oxidative stress tolerance, was studied by biochemical analyses and by electron paramagnetic resonance (EPR) technique. Whole grains as well as their parts: embryo, endosperm and seed coat were subjected to ozone treatment for 30 min. Biochemical investigation of control samples showed that their antioxidant activity increased in order: wheat<oat<barley. EPR method revealed that character and the number of paramagnetic species (transition metal ions: Fe(III), Cu(II), Mn(II) and stable organic radicals) changed upon ozone exposure, depending on the kind of cereal, stress tolerance of particular genotype and the part of grain. The control samples of whole grains and their parts originating from sensitive genotypes contained higher amounts of stable organic radicals (semiquinone, phenoxyl and carbohydrate types) than those from tolerant ones. In embryos of grains from sensitive genotypes their amount increased upon ozone treatment stronger than in embryos from grains of tolerant cultivars. In seed coats and endosperms such relation was not found and the changes in the content of the radicals during ozone application were correlated with the amount of transition metal ions and were more intensive in parts of grains richer in easily oxidized iron species Fe(II), located in inorganic structures. On the contrary, Fe(II) ions situated in embryos were stabilized by organic matrix and did not undergo oxidation by ozone.

Keywords: Cereal grains; EPR Spectroscopy; Ozone treatment; Stable organic radicals; Stress tolerance; Transition metal ions.