Efficient Agrobacterium tumefaciens-mediated transformation system of Diaporthe caulivora

MARINA MONTOYA; MASSA GABRIELA ALEJANDRA; MABEL COLABELLI; AZUCENA DEL CARMEN RIDAO

JOURNAL OF MICROBIOLOGICAL METHODS

ELSEVIER SCIENCE BV

Lugar: Amsterdam; Año: 2021

0167-7012

This is the first report describing the genetic transformation of Diaporthe caulivora, the soybean stem cankerfungus. A simple and 100 % efficient protocol of Agrobacterium tumefaciens-mediated transformation used mycelium asstarting 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 reactionwith hygromycin B resistance primers confirmed successful T-DNA integration into the fungal genome. Plant-fungusinteraction studies, including pathogenicity, latency, and endophytism, as well as further studies of random and targetedmutagenesis will be possible with GFP-expressing isolates of D. caulivora and other species in the Diaporthe / Phomopsiscomplex.