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Poster

Session: Session 1 Presenting Authors at Posters (Evens)

P-194 - Novel yeast-based polycistronic plasmids for the heterologous production of fungal secondary metabolites

Tuesday, July 15, 2025
09:55 - 11:30  
Location: Confex Hall 3
Microbial infection strategies (pathogens & non-pathogens)

    Author(s)

  • AG

    Aude Geistodt-Kiener

    Universite Paris-Saclay, INRAE, UR BIOGER
    Palaiseau, Ile-de-France, France

  • JT

    Jean Chrisologue Totozafy

    Universite Paris-Saclay, INRAE, AgroParisTech, IJPB
    Versailles, Ile-de-France, France

  • GL

    Géraldine Le Goff

    Universite Paris-Saclay, CNRS, ICSN
    Gif-sur-Yvette, Ile-de-France, France

  • KS

    Kaori Sakai

    Universite Paris-Saclay, INRAE, UR BIOGER
    Palaiseau, Ile-de-France, France

  • MV

    Muriel Viaud

    Group Leader team ECCP
    Universite Paris-Saclay, INRAE, UR BIOGER
    Palaiseau, Ile-de-France, France

  • RO

    Richard J. O'Connell

    Scientific Advisor of IMAFUN
    Université Paris-Saclay, INRAE, UR BIOGER
    Palaiseau, Ile-de-France, France

    • Presenting Author(s)

  • JD

    Jean-Felix Dallery

    Group Leader
    Universite Paris-Saclay, INRAE, UR BIOGER
    Palaiseau

Fungal pathogens of plants activate the expression of numerous biosynthetic gene clusters (BGC) exclusively when in the presence of a living host plant. This is a major bottleneck to the identification and structural elucidation of the corresponding fungal secondary metabolites. The heterologous expression of BGCs in microbial cell factories is one of the many approaches to get around such obstacles. We have implemented a system based on different Saccharomyces cerevisiae chassis strains and polycistronic vectors for efficient, seamless and cost-effective cloning of biosynthetic genes using in vivo assembly (also called transformation-assisted recombination) directly in Escherichia coli. Polycistrons were placed under the control of auto-inducible yeast promoters, pADH2 and pPCK1. As a proof-of-concept, we applied our system to the Colletochlorins, a family of phytotoxic molecules produced by Colletotrichum higginsianum. Using comparative genomics, we identified a BGC putatively involved in the biosynthesis of Colletochlorins in C. higginsianum, the causal agent of anthracnose disease of Brassicaceae. Expression of those genes in S. cerevisiae resulted in the identification of the precursor Orsellinic acid, Colletochlorins and their non-chlorinated counterparts, the Colletorins and allowed to link these compounds to their corresponding BGC. This system will now enable the production and purification of infection-specific secondary metabolites of fungal phytopathogens.