title: Reliable transformation system for Microbotryum lychnidis-dioicae informed by genome and transcriptome project.
abstract: Microbotryum lychnidis-dioicae is a member of a species complex infecting host plants in the Caryophyllaceae. It is used as a model system in many areas of research, but attempts to make this organism tractable for reverse genetic approaches have not been fruitful. Here, we exploited the recently obtained genome sequence and transcriptome analysis to inform our design of constructs for use in Agrobacterium-mediated transformation techniques currently available for other fungi. Reproducible transformation was demonstrated at the genomic, transcriptional and functional levels. Moreover, these initial proof-of-principle experiments provide evidence that supports the findings from initial global transcriptome analysis regarding expression from the respective promoters under different growth conditions of the fungus. The technique thus provides for the first time the ability to stably introduce transgenes and over-express target M. lychnidis-dioicae genes.
authors.affiliation: Department of Biology and Program on Disease Evolution, University of Louisville, Louisville, KY, 40292, USA.
authors.forename: Su San
authors.initials: SS
authors.lastname: Toh
authors.affiliation: Indiana University Southeast, New Albany, IN, USA.
authors.forename: David S
authors.initials: DS
authors.lastname: Treves
authors.affiliation: Kidney Disease Program, University of Louisville, Louisville, KY, USA.
authors.forename: Michelle T
authors.initials: MT
authors.lastname: Barati
authors.affiliation: Department of Biology and Program on Disease Evolution, University of Louisville, Louisville, KY, 40292, USA. mhperl01@louisville.edu.
authors.forename: Michael H
authors.identifier: http://orcid.org/0000-0002-5580-188X
authors.initials: MH
authors.lastname: Perlin
mesh_terms: D001487:Basidiomycota / Q000235:genetics*; D002874:Chromosome Mapping; D020869:Gene Expression Profiling; D018014:Gene Transfer Techniques; D016681:Genome, Fungal / Q000235:genetics*; D023281:Genomics; D010935:Plant Diseases / Q000382:microbiology*; D010944:Plants / Q000382:microbiology*; D059467:Transcriptome / Q000235:genetics; D014170:Transformation, Genetic / Q000235:genetics*
keywords: Agrobacterium-mediated transformation; Microbotryum; Recalcitrant fungi; Regulatable promoters