title: Multi-sited mutations of beta-tubulin are involved in benzimidazole resistance and thermotolerance of fungal biocontrol agent Beauveria bassiana.
abstract: Fungicide resistance and thermotolerance of biocontrol agents in mitosporic fungi are of merits for enhancing fungal formulations against insect pests in the field. Among 20 wild strains of Beauveria bassiana (a well-known fungal biocontrol agent) tested in this study, 19 were sensitive or highly sensitive to carbendazim (methyl 2-benzimidazole carbamate), a typical benzimidazole fungicide, despite low resistance found in one strain. Sequential mutagenesis of a carbendazim-sensitive wild strain [minimal inhibitory concentration (MIC) = 1.32 microg ml(-1)] under artificial selection pressure generated 11 mutants sharing a common MIC of > 1000 microg ml(-1) without visible variation in colony growth and conidiation capacity. This represents at least 758-fold enhancement of the resistance among the mutants. However, accompanied with the enhanced resistance, all the mutants became less thermotolerable. Stressed at 48 degrees C, conidial LT(50)s of the mutants varied from 1.8 to 9.6 min and were lower than the parental LT(50) (36 min). Moreover, the contents of hydrophobin-like proteins in conidial walls declined significantly among the mutants compared with that of the wild parent. Mutations commonly relating to benzimidazole resistance in fungi were located at Q134, F167 and/or E198 around the taxol-binding site of beta-tubulin by sequencing the beta-tubulin of the mutants. Also, mutations of other 37 amino acid residues in the sequences (each having one to five residues mutated) were found for the first time and they were diverse in spatial structure. All mutations restricted to the half of beta-tubulin close to alpha-tubulin were likely involved in variation in each of the traits concerned but their interactions were complicated.
authors.affiliation: Institute of Microbiology, College of Life Sciences, Zhejiang University, Hangzhou 310058, People's Republic of China.
authors.forename: Gen
authors.initials: G
authors.lastname: Zou
authors.forename: Sheng-Hua
authors.initials: SH
authors.lastname: Ying
authors.forename: Zhi-Chen
authors.initials: ZC
authors.lastname: Shen
authors.forename: Ming-Guang
authors.initials: MG
authors.lastname: Feng
mesh_terms: D000222:Adaptation, Physiological; D000818:Animals; D000871:Anthelmintics / Q000494:pharmacology*; D052982:Beauveria / Q000187:drug effects / Q000235:genetics*; D001562:Benzimidazoles / Q000494:pharmacology*; D004252:DNA Mutational Analysis; D019020:Directed Molecular Evolution / Q000009:adverse effects*; D004351:Drug Resistance; D025141:Drug Resistance, Fungal / Q000235:genetics*; D006376:Helminths / Q000382:microbiology; D006358:Hot Temperature / Q000009:adverse effects*; D010572:Pest Control, Biological; D014404:Tubulin / Q000187:drug effects / Q000235:genetics*
chemical_list: D000871:Anthelmintics; D001562:Benzimidazoles; D014404:Tubulin; C031000:benzimidazole