Visualisation of Reactive Oxygen Species During Stress of Aromatic Crop

The present protocol described staining protocol for Reactive Oxygen Species (ROS) in aromatic crop grown under nutrient stress through DAB and NBT histochemical method. Spearmint (Mentha spicata) were grown under manganese and salt toxicity stresses and after 10 and 20 days of the stress treatments, plants showed stunted growth. The morphological characteristics of leaves under stresses were observed. Manganese toxicity and salt stress induced the production of ROS. Accumulation of hydrogen peroxide was characterised as brown spots from the DAB polymerisation which were emerged and clearly observed in leaves from plant grown under 2.5 and 5 mM concentrations of manganese as well as 300 mM concentration of salt. Furthermore, accumulation of superoxide anion was characterised as blue pigments based upon the ability of cells to reduce NBT. Spearmint leaves showed the distribution of the blue pigment which was obviously observed under the 5 mM of manganese and 300 mM of salt. DAB and NBT staining method can be the rapid method to characterise ROS accumulation in plant cell under the abiotic stresses.