Efficient Agrobacterium tumefaciens-mediated transformation system of Diaporthe caulivora

J Microbiol Methods. 2021 May:184:106197. doi: 10.1016/j.mimet.2021.106197. Epub 2021 Mar 10.

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.

Keywords: GFP; Hygromycin; Latent infection; Soybean stem canker.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

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

Substances

  • Green Fluorescent Proteins
  • Hygromycin B

Supplementary concepts

  • Diaporthe caulivora