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Detection of Hydrogen Peroxide by DAB Staining in Arabidopsis Leaves
Arsalan Daudi1, Jose A O'Brien2
1Department of Biological Sciences, Royal Holloway University of London, Egham, UK; Department of Plant Pathology, University of California, Davis, CA, USA.
Abstract:
In this protocol, the in situ detection of hydrogen peroxide (one of several reactive oxygen species) is described in mature Arabidopsis rosette leaves by staining with 3,3'-diaminobenzidine (DAB) using an adaptation of previous methods (Thordal-Christensen et al., 1997; Bindschedler et al., 2006; Daudi et al., 2012). DAB is oxidized by hydrogen peroxide in the presence of some haem-containing proteins, such as peroxidases, to generate a dark brown precipitate. This precipitate is exploited as a stain to detect the presence and distribution of hydrogen peroxide in plant cells. The protocol can be modified slightly to detect hydrogen peroxide in different types of plant tissue.
