Triple-quad LC-MS-MS analysis for the detection of low-abundance plant signalling molecules in complex matrices

With the advent of genomics in plant science, the last decade our understanding of the genetic regulation of many processes in plants has strongly advanced. The high sensitivity of detection tools such as promoter reporter fusions, for example, allowed scientists to show that genetic regulation is often localised in specific tissues rather than occurring uniformly throughout entire organs. This implies that also the translation of the genetic regulation into a plant response probably occurs highly localised. Examples of this translation are the regulation of production and transport of signalling molecules such as plant hormones and other signalling molecules that are responsible for plant growth and the interaction with (organisms in) the environment. In order to be able to analyse local changes in the production of these signalling molecules, ultra-sensitive and highly selective analytical techniques are required that can detect specific molecules in complex matrices of hundreds of other metabolites. Hereto, we will use a technique called Multiple Reaction Monitoring (MRM) using Ultra Performance Liquid Chromatography- ElectroSpray Ionisation-tandem Mass Spectrometry UPLC/ESI-MS/MS. In this method we use the fact that compounds have a distinct precursor-to-product ion transition that is diagnostic for the presence of that compound in an extract. With the proposed Waters UPLC - Quatro Premier XE Mass Spectrometer we will analyse localised changes in plant hormones and the production of (rhizosphere) signalling molecules in a number of biological systems and as such contribute to the further elucidation of the regulatory processes in these systems. There is no comparable equipment present in any of the Dutch plant science research groups and for their questions with regard to plant hormones and plant signalling molecules, such as of importance in systemic acquired resistance, sugar signalling, shade avoidance and fruit development, these research groups now largely depend on foreign research groups. Also for them the proposed equipment is of great strategic importance and they therefore strongly support this initiative. Hence, the proposed equipment will allow Dutch plant scientists to further improve their understanding of the regulation of plant growth, plant development and plant defense. This improved knowledge is anticipated to lead to new patent applications, the improvement of agriculture, healthier foods and agricultural systems with lower inputs.