Consecutive action of two BAHD acyltransferases promotes tetracoumaroyl spermine accumulation in chicory - Plateforme Analytique Accéder directement au contenu
Article Dans Une Revue Plant Physiology Année : 2022

Consecutive action of two BAHD acyltransferases promotes tetracoumaroyl spermine accumulation in chicory

Julie Buges
David Mathiron
Jean-Louis Hilbert

Résumé

Fully substituted phenolamide accumulation in the pollen coat of Eudicotyledons is a conserved evolutionary chemical trait. Interestingly, spermidine derivatives are replaced by spermine derivatives as the main phenolamide accumulated in the Asteraceae family. Here, we show that the full substitution of spermine in chicory (Cichorium intybus) requires the successive action of two enzymes, i.e. CiSHT1 and CiSHT2 (spermidine hydroxycinnamoyl transferase-like protein 1 and 2), two members of the BAHD enzyme family. Deletion of these genes in chicory using CRISPR/Cas9 gene editing technology evidenced that CiSHT2 catalyzes the first N-acylation steps whereas CiSHT1 fulfills the substitution to give rise to tetracoumaroyl spermine. Additional experiments using Nicotiana benthamiana confirmed these findings. Expression of CiSHT2 alone promoted partially substituted spermine accumulation, and coexpression of CiSHT2 and CiSHT1 promoted synthesis and accumulation of the fully substituted spermine. Structural characterization of the main product of CiSHT2 using NMR revealed that CiSHT2 preferentially catalyzed N-acylation of secondary amines to form N5,N10-dicoumaroyl spermine, whereas CiSHT1 used this substrate to synthesize tetracoumaroyl spermine. We showed that spermine availability may be a key determinant towards preferential accumulation of spermine derivatives over spermidine derivatives in chicory. Our results reveal a subfunctionalization among the SHTs that was accompanied by a modification of free polyamine metabolism that has resulted in the accumulation of this new phenolamide in chicory and most probably in all Asteraceae. Finally, genetically engineered yeast (Saccharomyces cerevisiae) was shown to be a promising host platform to produce these compounds.
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Origine : Publication financée par une institution

Dates et versions

hal-03692347 , version 1 (20-01-2023)

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Paternité - Pas d'utilisation commerciale - Pas de modification

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Guillaume Bernard, Julie Buges, Marianne Delporte, Roland Molinié, Sebastien Besseau, et al.. Consecutive action of two BAHD acyltransferases promotes tetracoumaroyl spermine accumulation in chicory. Plant Physiology, 2022, ⟨10.1093/plphys/kiac234⟩. ⟨hal-03692347⟩
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