Abstract
The production of hydrogen peroxide (H2O2) has been recognized as an important feature of plant cells that undergo programmed cell death (PCD) during host-pathogen interaction. Thordal-Christensen et al. (1997) first described a method using chemical 3,3-diaminobenzidine (DAB) to detect the presence and distribution of H2O2 in barley leaves challenged by the powdery mildew fungus (Thordal-Christensen et al., 1997). Since then, this method has been adapted to many other plant species for in situ detection of H2O2. Here, we describe a modified protocol to stain and visualize H2O2 production in wheat leaves during infection by the necrotrophic fungus, Stagonospora nodrum or infiltration by the necrotrophic effectors produced by the fungus. The short version of this method has been reported in (Liu et al. 2012).
Materials and Reagents
Equipment
Procedure
Acknowledgments
This protocol was adapted from a method published in Liu et al. (2012), which was originally based on the descriptions by Thordal-Christensen et al. (1997). Development and implementation of this protocol was funded by USDA-NIFA AFRI Microbial Biology Program - Competitive grant # 2010-65108-20543, and by USDA-ARS CRIS Project #5442-22000-048-0D.
References
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