Agrobacterium tumefaciens-mediated transformation of Coniella granati

J Microbiol Methods. 2021 Mar:182:106149. doi: 10.1016/j.mimet.2021.106149. Epub 2021 Jan 22.

Abstract

Pomegranate fruit rot caused by Coniella granati is among the most devastating diseases threatening pomegranate production. The pathogenic mechanism of this pathogen remains largely unknown due to lack of genetic transformation method. Herein, we developed an approach to the Agrobacterium tumefaciens-mediated transformation (ATMT) of C. granati using a plasmid vector encoding the green fluorescent protein (GFP) and hygromycin resistance (Hyg) genes. This approach yielded C. granati transformants that exhibited uniform, stable green fluorescence. We further optimized this ATMT protocol, enabling us to achieve a transformation efficiency of up to 300 transformants per 0.5 cm2 mycelial plug. Together, we thus provide the first report of the stable transformation of C. granati, laying a foundation for future functional studies characterizing this economically important fungal pathogen.

Keywords: Agrobacterium tumefaciens-mediated transformation; Coniella granati; GFP; Hyg; Pomegranate fruit rot.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Ascomycota* / genetics
  • Ascomycota* / growth & development
  • DNA, Bacterial / genetics
  • Genetic Vectors
  • Green Fluorescent Proteins / genetics
  • Mutagenesis, Insertional
  • Mycoses / microbiology*
  • Plant Diseases / microbiology*
  • Pomegranate / microbiology*
  • Transformation, Genetic

Substances

  • DNA, Bacterial
  • Green Fluorescent Proteins

Supplementary concepts

  • Coniella granati