title: Involvement of the ABC transporter BcAtrB and the laccase BcLCC2 in defence of Botrytis cinerea against the broad-spectrum antibiotic 2,4-diacetylphloroglucinol.
abstract: The genetic and biochemical basis of defence mechanisms in plant pathogenic fungi against antifungal compounds produced by antagonistic microorganisms is largely unknown. The results of this study show that both degradative and non-degradative defence mechanisms enable the plant pathogenic fungus Botrytis cinerea to resist the broad-spectrum, phenolic antibiotic 2,4-diacetylphloroglucinol (2,4-DAPG). The efflux pump BcAtrB provides the first line of defence for B. cinerea, preventing accumulation of 2,4-DAPG in the cell to toxic concentrations, whereas the extracellular laccase BcLCC2 mediates, via conversion of tannic acid, subsequent degradation of 2,4-DAPG. Expression of BcatrB is induced by 2,4-DAPG and efflux gives B. cinerea sufficient time to more effectively initiate the process of BcLCC2-mediated antibiotic degradation. This is supported by the observations that the BcatrB mutant is significantly more sensitive to 2,4-DAPG than its parental strain, and is substantially less effective in 2,4-DAPG degradation. The results of this study further showed that BcLCC2 itself is not able to degrade 2,4-DAPG, but requires tannic acid as a mediator for 2,4-DAPG degradation. To our knowledge, this is the first time that the laccase-mediator system is shown to play a role in the detoxification of a broad-spectrum antibiotic compound from bacterial origin. We postulate that yet unknown constituents present in tannic acid act as substrate(s) of BcLCC2, thereby generating radicals that mediate 2,4-DAPG degradation.
authors.affiliation: Laboratory of Phytopathology, Wageningen University, PO Box 8025, 6700 EE Wageningen, The Netherlands. aschout@uni-bonn.de
authors.forename: Alexander
authors.initials: A
authors.lastname: Schouten
authors.forename: Olesya
authors.initials: O
authors.lastname: Maksimova
authors.forename: Yaite
authors.initials: Y
authors.lastname: Cuesta-Arenas
authors.forename: Grardy
authors.initials: G
authors.lastname: van den Berg
authors.forename: Jos M
authors.initials: JM
authors.lastname: Raaijmakers
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000235:genetics / Q000378:metabolism*; D000935:Antifungal Agents / Q000378:metabolism / Q000494:pharmacology*; D020171:Botrytis / Q000187:drug effects* / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D003470:Culture Media; D025141:Drug Resistance, Fungal; D042845:Laccase / Q000235:genetics / Q000378:metabolism*; D008826:Microbial Sensitivity Tests; D009154:Mutation; D010696:Phloroglucinol / Q000031:analogs & derivatives / Q000378:metabolism / Q000494:pharmacology
chemical_list: C437164:ABC transporter BcatrB; D018528:ATP-Binding Cassette Transporters; D000935:Antifungal Agents; D003470:Culture Media; C059817:2,4-diacetylphloroglucinol; D010696:Phloroglucinol; D042845:Laccase