title: Agrobacterium tumefaciens-mediated transformation of the lichen fungus, Umbilicaria muehlenbergii.
abstract: Transformation-mediated mutagenesis in both targeted and random manners has been widely applied to decipher gene function in diverse fungi. However, a transformation system has not yet been established for lichen fungi, severely limiting our ability to study their biology and mechanism underpinning symbiosis via gene manipulation. Here, we report the first successful transformation of the lichen fungus, Umbilicaria muehlenbergii, via the use of Agrobacterium tumefaciens. We generated a total of 918 transformants employing a binary vector that carries the hygromycin B phosphotransferase gene as a selection marker and the enhanced green fluorescent protein gene for labeling transformants. Randomly selected transformants appeared mitotically stable, based on their maintenance of hygromycin B resistance after five generations of growth without selection. Genomic Southern blot showed that 88% of 784 transformants contained a single T-DNA insert in their genome. A number of putative mutants affected in colony color, size, and/or morphology were found among these transformants, supporting the utility of Agrobacterium tumefaciens-mediated transformation (ATMT) for random insertional mutagenesis of U. muehlenbergii. This ATMT approach potentially offers a systematic gene functional study with genome sequences of U. muehlenbergii that is currently underway.
authors.affiliation: Korean Lichen Research Institute, Sunchon National University, Sunchon, Korea ; Dept. of Agricultural Biotechnology, Fungal Bioinformatics Laboratory, Center for Fungal Genetic Resources, and Center for Fungal Pathogenesis, Seoul National University, Seoul, Korea.
authors.forename: Sook-Young
authors.initials: SY
authors.lastname: Park
authors.affiliation: Korean Lichen Research Institute, Sunchon National University, Sunchon, Korea ; Dept. of Biology, Sunchon National University, Sunchon, Korea.
authors.forename: Min-Hye
authors.initials: MH
authors.lastname: Jeong
authors.affiliation: College of Life Sciences, Shandong Normal University, Jinan, China.
authors.forename: Hai-Ying
authors.initials: HY
authors.lastname: Wang
authors.affiliation: Korean Lichen Research Institute, Sunchon National University, Sunchon, Korea.
authors.forename: Jung A
authors.initials: JA
authors.lastname: Kim
authors.affiliation: Korean Lichen Research Institute, Sunchon National University, Sunchon, Korea ; Dept. of Biology, Sunchon National University, Sunchon, Korea.
authors.forename: Nan-Hee
authors.initials: NH
authors.lastname: Yu
authors.affiliation: Dept. of Agricultural Biotechnology, Fungal Bioinformatics Laboratory, Center for Fungal Genetic Resources, and Center for Fungal Pathogenesis, Seoul National University, Seoul, Korea.
authors.forename: Sungbeom
authors.initials: S
authors.lastname: Kim
authors.affiliation: Korean Lichen Research Institute, Sunchon National University, Sunchon, Korea.
authors.forename: Yong Hwa
authors.initials: YH
authors.lastname: Cheong
authors.affiliation: Dept. of Plant Pathology & Environmental Microbiology, The Pennsylvania State University, University Park, Pennsylvania, United States of America.
authors.forename: Seogchan
authors.initials: S
authors.lastname: Kang
authors.affiliation: Dept. of Agricultural Biotechnology, Fungal Bioinformatics Laboratory, Center for Fungal Genetic Resources, and Center for Fungal Pathogenesis, Seoul National University, Seoul, Korea.
authors.forename: Yong-Hwan
authors.initials: YH
authors.lastname: Lee
authors.affiliation: Korean Lichen Research Institute, Sunchon National University, Sunchon, Korea.
authors.forename: Jae-Seoun
authors.initials: JS
authors.lastname: Hur
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics*; D001203:Ascomycota / Q000235:genetics* / Q000302:isolation & purification; D004269:DNA, Bacterial / Q000235:genetics; D005800:Genes, Fungal / Q000235:genetics; D005818:Genetic Engineering / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000235:genetics; D009154:Mutation; D010641:Phenotype; D014170:Transformation, Genetic
chemical_list: D004269:DNA, Bacterial; C036483:T-DNA; D049452:Green Fluorescent Proteins