KBC/EMORR Experimental Methods in Studies of Plant Defense Responses

Guarantor: Marek Petřivalský
Tutorial lecturer: Lucie Kubienová
Exercise: 8 hours/week
Credits: 4
Winter semester
Form of course completion: course credit

Individual laboratory activities include protocols of plant material preparation and practical design of various abiotic and biotic stress treatments, as well as plant sample preparation for following analysis by spectrophotometric, fluorimetric, chromatographic, electrophoretic and histochemical methods:

• Preparation of plant material. Stress factors
• Purification of plant protoplasts and plant cell culture
• Analysis of cell density, viability, cell death and plat cell wall regeneration
• Analysis and localization of reactive oxygen species
• Analysis of reactive nitrgen species in plant extrakts
• Localization of nitric oxide production in plant tissue
• Determination of lipid peroxidation in plant extrakts
• Determination of total antioxidant capacity in plant extrakts
• Determination of enzyme activities – ascorbate peroxidase, glutathione peroxidase and catalasetr
• Determination of isoenzymes o plant peroxidase by native PAGE
• Determination of activity of phenylalanine amoniak lyase. Localization of phenolic substances
• Determination of free and bounded phenolic substances
• Determination of free and conjugated polyamines by HPLC
  Analysis of low molecular and protein S-nitrosothiols in planT extrakts