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J Microbiol Methods ; 184: 106197, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33713724

RESUMO

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.


Assuntos
Agrobacterium tumefaciens/genética , Ascomicetos/genética , Técnicas de Transferência de Genes , Transformação Genética , Agrobacterium tumefaciens/metabolismo , Ascomicetos/efeitos dos fármacos , Ascomicetos/metabolismo , Vetores Genéticos/genética , Vetores Genéticos/metabolismo , Genoma Fúngico , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Higromicina B/farmacologia , Phomopsis/genética , Doenças das Plantas/microbiologia , Glycine max/microbiologia
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