title: Agrobacterium tumefaciens-mediated transformation in the entomopathogenic fungus Lecanicillium lecanii and development of benzimidazole fungicide resistant strains.
abstract: Lecanicillium lecanii has been used in the biological control of several insects in agricultural practice. Since the gene manipulation tools for this entomopathogenic fungus have not been sufficiently developed, Agrobacterium tumefaciens-mediated transformation (ATMT) in L. lecanii was investigated in this study, using the wild-type isolate FZ9906 as a progenitor strain and the hygromycin B resistance (hph) gene as a selection marker. Furthermore, a field carbendazim-resistant (mrt) gene from Botrytis cinerea was expressed in L. lecanii FZ9906 via the ATMT system. The results revealed that the frequency of transformation surpassed 25transformants/10(6) conidia, most of the putative transformants contained a single copy of T-DNA, and the T-DNA inserts were stably inherited after five generations. All putative transformants had indistinguishable biological characteristics relative to the wild-type strain, excepting two transformants with altered growth habits or virulence. Moreover, the resistance of the putative transformants to carbendazim (MBC) was improved, and the highest one was 380-fold higher than the wild-type strain. In conclusion, ATMT is an effective and suitable system for L. lecanii transformation, and will be a useful tool for the basic and application research of gene functions and gene modifications of this strain.
authors.affiliation: State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, People's Republic of China.
authors.forename: Yan-Jun
authors.initials: YJ
authors.lastname: Zhang
authors.affiliation: State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, People's Republic of China.
authors.forename: Jin-Jin
authors.initials: JJ
authors.lastname: Zhao
authors.affiliation: State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, People's Republic of China. Electronic address: xiem406@126.com.
authors.forename: Ming
authors.initials: M
authors.lastname: Xie
authors.affiliation: State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, People's Republic of China.
authors.forename: De-Liang
authors.initials: DL
authors.lastname: Peng
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics; D000935:Antifungal Agents / Q000494:pharmacology*; D001562:Benzimidazoles / Q000494:pharmacology*; D032661:Cordyceps / Q000187:drug effects* / Q000235:genetics*; D025141:Drug Resistance, Fungal; D006921:Hygromycin B / Q000494:pharmacology; D060847:Metabolic Engineering / Q000379:methods*; D012641:Selection, Genetic; D014170:Transformation, Genetic
chemical_list: D000935:Antifungal Agents; D001562:Benzimidazoles; D006921:Hygromycin B; C031000:benzimidazole
keywords: Entomopathogenic fungus; Fungicide resistance; Genetic transformation