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Citronellol biosynthesis in pelargonium is a multi-step pathway involving PRISE enzymes.

Laure MartinelliCamille BihanicAurélie BonyFlorence GrosCorentin ConartSébastien FiorucciHervé CasabiancaFrédéric SchietsGiorgiana ChieteraBenoît BoachonBernard BlerotSylvie BaudinoFrédéric JullienDenis Saint-Marcoux
Published in: Plant physiology (2023)
Citronellol is a pleasant-smelling compound produced in rose (Rosa spp.) flowers and in the leaves of many aromatic plants, including pelargoniums (Pelargonium spp.). Although geraniol production has been well studied in several plants, citronellol biosynthesis has been documented only in crab-lipped spider orchid (Caladenia plicata) and its mechanism remains open to question in other species. We therefore profiled 10 pelargonium accessions using RNA-Seq and GC-MS analysis. Three enzymes from the PRISE family were characterized in vitro and subsequently identified as citral reductases (named PhCIRs). Transgenic RNAi lines supported a role for PhCIRs in the biosynthesis of citronellol as well as in the production of mint-scented terpenes. Despite their high amino acid sequence identity, the three enzymes showed contrasting stereoselectivity, either producing mainly (S)-citronellal or a racemate of both (R)- and (S)-citronellal. Using site-directed mutagenesis, we identified a single amino acid substitution as being primarily responsible for the enzyme's enantioselectivity. Phylogenetic analysis of pelargonium PRISEs revealed three clades and seven groups of orthologs. PRISEs from different groups exhibited differential affinities toward substrates (citral and progesterone) and cofactors (NADH/NADPH), but most were able to reduce both substrates, prompting hypotheses regarding the evolutionary history of PhCIRs. Our results demonstrate that pelargoniums evolved citronellol biosynthesis independently through a three-step pathway involving PRISE homologues and both citral and citronellal as intermediates. In addition, these enzymes control the enantiomeric ratio of citronellol thanks to small alterations of the catalytic site.
Keyphrases
  • amino acid
  • rna seq
  • single cell
  • cell wall
  • minimally invasive
  • crispr cas
  • genome wide
  • reactive oxygen species
  • dna methylation
  • mass spectrometry
  • genetic diversity
  • solid state
  • wild type