Automated image analysis for quantification of histochemical detection of reactive oxygen species and necrotic infection symptoms in plant leaves

Abstract

In plants, the accelerated production of reactive oxygen species (ROS) is a common response to different stresses. We developed a computer method of leaf image analysis to automatically quantify the and H₂O₂ staining patterns in leaves. The quantification of histochemically detected ROS is given as a percentage of stained area partitioned according to the staining intensity and hence accumulation. Thus, this method expands the routine histochemical detection of ROS in situ. The same procedure can be also used to assess necrotic infection symptoms. In plants infected with necrotizing pathogens leaves can be simultaneously examined for pathogen-induced and H₂O₂ accumulation and infection development. This procedure allows rapid and accurate examination of many leaves and provides an advantage over the routine visual assessment of in situ histochemical detection of ROS and infection symptom development.

Keywords:

• histochemistry
• hydrogen peroxide
• infection symptoms
• leaf image analysis
• superoxide anion radical