Lead-induced stress, which triggers the production of nitric oxide (NO) and superoxide anion (O2·-) in Arabidopsis peroxisomes, affects catalase activity

Abstract

Lead (Pb) contamination has a toxic effect on plant metabolisms, leading to a decrease in biomass production. The free radical nitric oxide (NO) is involved in the mechanism of response to a wide range of abiotic stresses. However, little is known about the interplay between Pb-induced stress and NO metabolism. Peroxisomes are sub-cellular compartments involved in multiple cellular metabolic pathways which are characterized by an active nitro-oxidative metabolism. Thus, Arabidopsis thaliana mutants expressing cyan fluorescent protein (CFP) through the addition of peroxisomal targeting signal 1 (PTS1), which enables peroxisomes to be visualized in vivo by confocal laser scanning microscopy (CLSM) combined with fluorescent probes for nitric oxide (NO), superoxide anion (O2·-) and peroxynitrite (ONOO−), were used to evaluate the potential involvement of these organelles in the mechanism of response to 150 μM lead-induced stress. Both NO and O2·- radicals, and consequently ONOO−, were overproduced under Pb-stress. Additionally, biochemical and gene expression analyses of peroxisomal enzymes, including the antioxidant catalase (CAT) and two photorespiration enzymes, such as glycolate oxidase (GOX) and hydroxypyruvate reductase (HPR), show that, under Pb-stress, only the catalase was negatively affected, while the two photorespiration enzymes remained unaffected. These results corroborate the involvement of plant peroxisomal metabolisms in the mechanism of response to lead contamination and highlight the importance of the peroxisomal NO metabolism.