2.5.2017: Miroslav STRNAD (Olomouc, Czech Republic): Phytohormone profiling on tissue and cell level (Olomouc, Czech Republic): Phytohormone profiling on tissue and cell level, Hörsaal 31.11, Institut für Pflanzenwissenschaften, Bereich Pflanzenphysiologie, Schubertstraße 51, um 17:00 Uhr.
Plant hormones play crucial role in the control of various physiological processes. The major problem associated with phytohormone analysis is that the amount of phytohormones present endogenously in plant tissues is very low, usually in the range of fmol to pmol/g fresh weight. High-resolution measurements of phytohormones are therefore necessary for physiological studies of their mode of action. Application of targeted metabolomics shows an optimal method for phytohormonal screening. Our work is focused on the development of high-throughput miniaturized purification methods for minute amounts of plant tissue. Two novel approaches, a simple one-step purification protocol based on in-tip microSPE (micro Solid-Phase Extraction) and a class-specific miniaturized immunoaffinity chromatography method were utilized. To verify the effect of complex sample matrix as well as its efficiency and analytical accuracy, the appropriate stable isotope-labelled internal standards have been prepared by the organic synthesis. We have also developed several fast chromatographic separations (UHPLC) and highly sensitive tandem mass spectrometry (MS/MS) methods for simultaneous profiling of the phytohormone metabolites. Moreover, we applied fluorescence-activated cell sorting (FACS) of green fluorescent protein (GFP)-marked cell types, combined with a highly sensitive LC-MS method for analysis of phytohormonal biosynthesis and homeostasis at tissue resolution.