METHOD FOR PROFILING PHYTOHORMONE LEVELS IN PLANT TISSUE

摘要

The present invention provides a method for profiling phytohormone levels in plant tissue or tissue of other plastid containing organisms, i.e. a method for the simultaneous determination of a multitude of phytohormone levels in plant tissue or tissue of other plastid containing organisms, which method comprises the following steps: i. extraction of particulate tissue material of the plastid containing organism, in particulate tissue material of plants, with a liquid extractant, which is a mixture of at least one water miscible neutral organic solvent having from 1 to 3 carbon atoms and 1 heteroatom selected from O and N with water containing from 0.1 to 5% by weight, based on the extractant, of at least one acid, whereby a first liquid extract is obtained; ii. contacting the liquid extract obtained in step i. with a solid absorbent having a hydrophobically modified surface and removing the solid absorbent to obtain a second liquid extract; iii. evaporating the solvent from the second extract and then re-dissolving the obtained residue in a solvent mixture of at least one water miscible neutral organic solvent having from 1 to 3 carbon atoms and 1 heteroatom selected from O and N with water containing from 0.1 to 5% by weight, based on the solvent mixture, of at least one acid to obtain a reconstituted extract; iv. purification of the reconstituted extract by contacting it with a solid absorbent having a hydrophobically modified surface to obtain a purified reconstituted extract (eluate); and v. determining the relative concentrations of at least two, frequently at least 4, in particular at least 6, especially at least 8 or at least 10 phyto, e.g. from 2 to 60, frequently from 4 to 50, in particular from 6 to 45, especially from 8 to 40 or from 10 to 30 plant hormones (phytohormones) in the purified reconstituted extract obtained in step iv. by directly subjecting the purified reconstituted extract to an analyzing unit comprising a separation unit for separation the phytohormones and an analyzer for identifying the phytohormones which separation unit is coupled to the analyzer.