Electrochemical detection of extracellular hydrogen peroxide in Arabidopsis thaliana: A real-time marker of oxidative stress
Gonzalez-Sanchez, M. I. , Gonzalez-Macia, L. , Hancock, J. T. and Killard, A. (2013) Electrochemical detection of extracellular hydrogen peroxide in Arabidopsis thaliana: A real-time marker of oxidative stress. To be published in Plant, Cell and Environment, 36 (4). pp. 869-878. ISSN 0140-7791 [In Press]
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Publisher's URL: http://dx.doi.org/10.1111/pce.12023
An electrochemical approach to directly measure the dynamic process of H2O2 release from cultures of Arabidopsis thaliana cells is reported. This approach is based on H2O2 oxidation on a Pt electrode in conjunction with continuous measurement of sample pH. For [H2O2] < 1 mM, calibration plots were linear and the amperometric response of the electrode was dependent on the pH. At higher concentrations ([H2O2] > 1 mM), the amperometric response can be described by Michaelian-type kinetics and a mathematical expression relating current intensity and pH was obtained to quantitatively determine H2O2 concentration. At pH 5.5, the detection limit of the sensor was 3.1 M (S/N=3), with a response sensitivity of 0.16 AM-1cm-2 and reproducibility was within 6.1% in the range 1-5 x 10-3 M (n=5). Cell suspensions under normal physiological conditions had a pH between 5.5-5.7 and H2O2 concentrations in the range 7.0-20.5 M (n=5). The addition of exogenous H2O2, as well as other potential stress stimuli was made to the cells and the change in H2O2 concentration was monitored. This real-time quantitative H2O2 analysis is a potential marker for the evaluation of oxidative stress in plant cell cultures.
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