Sesquiterpene diversity and TPS-a gene characterization in the pelargonium genus

Sesquiterpenes are key molecules nurturing the complex and subtle base notes of pelargonium fragrances. Yet, their contribution to the essential oil olfactory profile and their biosynthesis are far from being well understood. We therefore explored sesquiterpene composition in 10 pelargonium accessions and revealed that, although lowly accumulated, their wide diversity participates to the biochemical uniqueness of each fragrance. We further investigated sesquiterpene biosynthesis thanks to a multiomic approach, integrating transcriptomic together with metabolomic data. A phylogenetic analysis of pelargonium TPS-a subfamily first revealed a total of 21 groups of orthology and denoted a strong transcriptional regulation of related enzymes. Seven sesquiterpene synthases were then wisely selected for functional characterization, both in vitro and in planta. Among them, two 6,9-guaidiene synthases were characterized for the first time in plants. Finally, we explored the role of amino acids located in the active site of sesquiterpene synthases in the enzymatic mechanism using site-directed mutagenesis. Altogether, this work highlights the great potential of the multiomic approach to predict TPS functions, but also exposes its limitations, inherent to terpene synthase's plasticity.