Analytical methods – determination of plant hormones
Understanding of the hormonal control of plant development is dependent on the application of appropriate methodological procedures. An independent analytical unit was established in our laboratory that deals with improvement of existing and development of new modern methods of plant hormone analysis, including their extraction, purification, identification, and metabolic profiling.
In cooperation with other members of the laboratory a new fast and efficient method of extraction and purification of several groups of plant hormones and other bioactive compounds from a single plant extract was developed (Dobrev and Kamínek, J. Chrom.A 2002, Dobrev et al., J. Chrom.A 2005 a Hoyerová et al., Phytochemistry 2006). The efficiency and relative simplicity of this method led to its quick spreading among plant hormone community as indicates its frequent citation (Dobrev and Kamínek, 2002, prized as „top cited paper“ in J. Chromatography A).
A new method for on line sample purification and quantitation of plant hormones auxin and abscissic acid utilizing two dimensional high performance liquid chromatography (2D-HPLC) was developed (Dobrev et al., J. Chrom.A 2005).
Methods were developed and optimized for determination of biological activities of key hormone enzymes catalyzing various metabolic conversions of cytokinins, such as cytokinin oxidase/dehydrogenase (Dobrev a Motyka, Biologické Listy 2003), zeatin reductase (Gaudinová et al., J. Plant Growth Regul. 2005) and cytokinin glucosyltransferase (Polanská et al., Biotechnology Letters 2004).
Metabolic profile of trans-zeatin in tobacco cells.
The analytical unit also deals with “metabolic profiling” of plant hormones and other bioactive compounds including studies of metabolic fate and dynamics of labeled analytes in plant cells.