title: Flexible gateway constructs for functional analyses of genes in plant pathogenic fungi.
abstract: Genetic manipulation of fungi requires quick, low-cost, efficient, high-throughput and molecular tools. In this paper, we report 22 entry constructs as new molecular tools based on the Gateway technology facilitating rapid construction of binary vectors that can be used for functional analysis of genes in fungi. The entry vectors for single, double or triple gene-deletion mutants were developed using hygromycin, geneticin and nourseothricin resistance genes as selection markers. Furthermore, entry vectors containing green fluorescent (GFP) or red fluorescent (RFP) in combination with hygromycin, geneticin or nourseothricin selection markers were generated. The latter vectors provide the possibility of gene deletion and simultaneous labelling of the fungal transformants with GFP or RFP reporter genes. The applicability of a number of entry vectors was validated in Zymoseptoria tritici, an important fungal wheat pathogen.
authors.affiliation: Cereal Research Department, Seed and Plant Improvement Institute, PO Box 31585-4119, Karaj, Iran; Laboratory of Phytopathology, Wageningen University, Wageningen, The Netherlands. Electronic address: Rahim.Mehrabi@gmail.com.
authors.forename: Rahim
authors.initials: R
authors.lastname: Mehrabi
authors.affiliation: Plant Research International B.V., Wageningen University and Research Centre, The Netherlands; Department of Plant Protection, College of Agriculture, University of Tehran, Plant Pathology Building, Karaj, Iran.
authors.forename: Amir
authors.initials: A
authors.lastname: Mirzadi Gohari
authors.affiliation: Laboratory of Molecular Biology, Embrapa Western Amazon, Manaus, Brazil.
authors.forename: Gilvan Ferreira
authors.initials: GF
authors.lastname: da Silva
authors.affiliation: School of Biosciences, University of Exeter, Exeter, UK.
authors.forename: Gero
authors.initials: G
authors.lastname: Steinberg
authors.affiliation: Plant Research International B.V., Wageningen University and Research Centre, The Netherlands. Electronic address: Gert.Kema@wur.nl.
authors.forename: Gert H J
authors.initials: GH
authors.lastname: Kema
authors.affiliation: Laboratory of Phytopathology, Wageningen University, Wageningen, The Netherlands.
authors.forename: Pierre J G M
authors.initials: PJ
authors.lastname: de Wit
mesh_terms: D025141:Drug Resistance, Fungal; D005658:Fungi / Q000235:genetics* / Q000472:pathogenicity / Q000502:physiology; D015870:Gene Expression; D018390:Gene Targeting / Q000379:methods*; D005800:Genes, Fungal; D005822:Genetic Vectors; D005827:Genetics, Microbial / Q000379:methods*; D008164:Luminescent Proteins / Q000032:analysis / Q000235:genetics; D008967:Molecular Biology / Q000379:methods*; D010944:Plants / Q000382:microbiology*; D012641:Selection, Genetic; D013194:Staining and Labeling / Q000379:methods; D014170:Transformation, Genetic
chemical_list: D008164:Luminescent Proteins
keywords: Binary vectors; GFP; Gateway technology; Gene deletion; RFP; Zymoseptoria tritici