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J Microbiol Methods ; 184: 106197, 2021 05.
Article in English | MEDLINE | ID: mdl-33713724

ABSTRACT

This is the first report describing the genetic transformation of Diaporthe caulivora, the soybean stem canker fungus. A simple and 100% efficient protocol of Agrobacterium tumefaciens-mediated transformation used mycelium as starting material and the hygromycin B resistance and green fluorescent protein (GFP) as a selection and reporter agents, respectively. All transgenic isolates were mitotically stable in two independent experiments and polymerase chain reaction with hygromycin B resistance primers confirmed successful T-DNA integration into the fungal genome. Plant-fungus interaction studies, including pathogenicity, latency, and endophytism, as well as further studies of random and targeted mutagenesis will be possible with GFP-expressing isolates of D. caulivora and other species in the Diaporthe / Phomopsis complex.


Subject(s)
Agrobacterium tumefaciens/genetics , Ascomycota/genetics , Gene Transfer Techniques , Transformation, Genetic , Agrobacterium tumefaciens/metabolism , Ascomycota/drug effects , Ascomycota/metabolism , Genetic Vectors/genetics , Genetic Vectors/metabolism , Genome, Fungal , Green Fluorescent Proteins/genetics , Green Fluorescent Proteins/metabolism , Hygromycin B/pharmacology , Phomopsis/genetics , Plant Diseases/microbiology , Glycine max/microbiology
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