Resveratrol acts as a natural profungicide and induces self-intoxication by a specific laccase.
The grapevine (Vitis) secondary metabolite resveratrol is considered a phytoalexin, which protects the plant from Botrytis cinerea infection. Laccase activity displayed by the fungus is assumed to detoxify resveratrol and to facilitate colonization of grape. We initiated a functional molecular genetic analysis of B. cinerea laccases by characterizing laccase genes and evaluating the phenotype of targeted gene replacement mutants. Two different laccase genes from B. cinerea were characterized, Bclcc1 and Bclcc2. Only Bclcc2 was strongly expressed in liquid cultures in the presence of either resveratrol or tannins. This suggested that Bclcc2, but not Bclcc1, plays an active role in the oxidation of both resveratrol and tannins. Gene replacement mutants in the Bclcc1 and Bclcc2 gene were made to perform a functional analysis. Only Bclcc2 replacement mutants were incapable of converting both resveratrol and tannins. When grown on resveratrol, both the wild type and the Bclcc1 replacement mutant showed inhibited growth, whereas Bclcc2 replacement mutants were unaffected. Thus, contrary to the current theory, BcLCC2 does not detoxify resveratrol but, rather, converts it into compounds that are more toxic for the fungus itself. The Bclcc2 gene was expressed during infection of B. cinerea on a resveratrol-producing host plant, but Bclcc2 replacement mutants were as virulent as the wild-type strain on various hosts. The activation of a plant secondary metabolite by a pathogen introduces a new dimension to plant-pathogen interactions and the phytoalexin concept.
Schouten
Alexander
A
Wageningen University Plant Sciences, Laboratory of Phytopathology, PO Box 8025, 6700 EE Wageningen, The Netherlands.
Wagemakers
Lia
L
Stefanato
Francesca L
FL
van der Kaaij
Rachel M
RM
van Kan
Jan A L
JA
D000595:Amino Acid Sequence; D000935:Antifungal Agents / Q000378:metabolism*; D010367:Arachis / Q000382:microbiology; D001483:Base Sequence; D020171:Botrytis / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D004271:DNA, Fungal; D015870:Gene Expression; D005800:Genes, Fungal; D047348:Hydrolyzable Tannins / Q000378:metabolism; D042845:Laccase; D008969:Molecular Sequence Data; D016296:Mutagenesis; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism*; D010636:Phenols / Q000378:metabolism*; D018515:Plant Leaves / Q000382:microbiology; D011355:Prodrugs / Q000378:metabolism*; D000077185:Resveratrol; D017386:Sequence Homology, Amino Acid; D013267:Stilbenes / Q000378:metabolism*; D014774:Virulence; D027843:Vitis / Q000382:microbiology
D000935:Antifungal Agents; D004271:DNA, Fungal; D047348:Hydrolyzable Tannins; D010636:Phenols; D011355:Prodrugs; D013267:Stilbenes; D010088:Oxidoreductases; D042845:Laccase; D000077185:Resveratrol