Reactive oxygen species (ROS) are by-products of photosynthesis and respiration in plant tissues. Abiotic and biotic stressors also induce the production and temporary accumulation of ROS in plants, whereby they can act as secondary messengers/chemical mediators in plant defense signaling and lead to programmed cell death. Hydrogen peroxide (H2O2), one of the key ROS, acts as central hub for information flow in plants. Despite such a key role in fundamental cellular processes, reliable quantitative methods for the determination of H2O2 levels in plant tissues are difficult to find. In this study we quantified constitutive and/or induced levels of H2O2 in a woody plant (pine, Pinus nigra), a crop plant (rice, Oryza sativa) and a model plant (Arabidopsis thaliana) using the fluorimetric probe Amplex Red. Lower limit of detection (LLD) and lower limit of quantitation (LLQ) were determined in the low picomole range. Overall, assay sensitivity for the plant extracts was in the nanomol/g FW range. To estimate the H2O2 scavenging capacity of each plant taxon, we spiked the respective extracts with known concentrations of H2O2 and measured the resulting fluorescence. Finally, we demonstrate that a working solution of Amplex Red remains stable for at least 30 days at -80 �C.

Hydrogen peroxide quantification in plants using the fluorimetric probe Amplex� Red

247th National Meeting (2014)

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