title: M233I Mutation in the β-Tubulin of Botrytis cinerea Confers Resistance to Zoxamide.
abstract: Three phenotypes were detected in 161 Botrytis cinerea field isolates, including Zox(S)Car(S) (sensitive to zoxamide and carbendazim), Zox(S)Car(R) (sensitive to zoxamide and resistant to carbendazim), and Zox(R)Car(R) (resistant to zoxamide and carbendazim), but not Zox(R)Car(S) (resistant to zoxamide and sensitive to carbendazim). The baseline sensitivity to zoxamide was determined with a mean EC50 of 0.76 μg/ml. Two stable Zox(R)Car(S) isolates were obtained with a resistance factor of 13.28 and 20.43; there was a fitness penalty in mycelial growth rate, sporulation, virulence and sclerotium production. The results suggest that the resistance risk of B. cinerea to zoxamide is low where benzimidazoles have not been used. E198V, E198K and M233I, were detected in the β-tubulin of Zox(S)Car(R), Zox(R)Car(R), Zox(R)Car(S), respectively. Molecular docking indicated that position 198 in β-tubulin were targets for both zoxamide and carbendazim. The mutations at 198 prevented formation of hydrogen bonds between β-tubulin and carbendazim (E198V/K), and changed the conformation of the binding pocket of zoxamide (E198K). M233I had no effect on the binding of carbendazim but resulted in loss of a hydrogen bond between zoxamide and F200. M233 is suggested to be a unique target site for zoxamide and be very important in the function of β tubulin.
authors.affiliation: Department of Plant Pathology, China Agricultural University, Beijing, 100193, P. R. China.
authors.forename: Meng
authors.initials: M
authors.lastname: Cai
authors.affiliation: Department of Plant Pathology, China Agricultural University, Beijing, 100193, P. R. China.
authors.forename: Dong
authors.initials: D
authors.lastname: Lin
authors.affiliation: College of Forestry, Beijing Forestry University, Beijing, 100083, P.R. China.
authors.forename: Lei
authors.initials: L
authors.lastname: Chen
authors.affiliation: Plant Science and Technology College, Beijing University of Agriculture, Beijing, 102206, P.R. China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Bi
authors.affiliation: Department of Plant Pathology, China Agricultural University, Beijing, 100193, P. R. China.
authors.forename: Lu
authors.initials: L
authors.lastname: Xiao
authors.affiliation: Department of Plant Pathology, China Agricultural University, Beijing, 100193, P. R. China.
authors.forename: Xi-li
authors.initials: XL
authors.lastname: Liu
mesh_terms: D000577:Amides / Q000737:chemistry / Q000494:pharmacology*; D001562:Benzimidazoles / Q000494:pharmacology; D001665:Binding Sites; D020171:Botrytis / Q000187:drug effects / Q000235:genetics*; D002219:Carbamates / Q000494:pharmacology; D025141:Drug Resistance, Fungal; D005659:Fungicides, Industrial / Q000737:chemistry / Q000494:pharmacology*; D008826:Microbial Sensitivity Tests; D062105:Molecular Docking Simulation; D020125:Mutation, Missense; D025282:Mycelium / Q000187:drug effects / Q000235:genetics; D010935:Plant Diseases; D011485:Protein Binding; D013172:Spores, Fungal / Q000187:drug effects / Q000235:genetics; D014404:Tubulin / Q000737:chemistry / Q000235:genetics*; D014774:Virulence
chemical_list: D000577:Amides; D001562:Benzimidazoles; D002219:Carbamates; D005659:Fungicides, Industrial; D014404:Tubulin; C451427:zoxamide; C006698:carbendazim