Artigo
Development of a transformation system for Crinipellis perniciosa, the causal agent of witches' broom in cocoa plants
Autor
Lima, Juliana Oliveira
Santos, Jildete Karla dos
Pereira, Jorge Fernando
Resende, Mário Lúcio Vilela de
Araújo, Elza Fernandes de
Queiroz, Marisa Vieira de
Institución
Resumen
Protoplasts of the pathogenic plant fungus, Crinipellis perniciosa, were transformed to hygromycin B resistance using the pAN7-1 plasmid, which contains the Escherichia coli hph gene under the control of Aspergillus nidulans regulatory sequences. The pAN7-1 plasmid was introduced by PEG/CaCl2 treatment. Transformation frequencies of 1.6–2.5 transformants/μg of DNA were achieved. About 54% of the transformants were abortive and 40 analyzed transformants were mitotically stable and showed different hygromycin B resistance levels. The presence of the hph gene was checked by PCR in five transformants and the integration of multiple plasmid copies into different genome sites was observed by Southern analysis. This is the first report of a C. perniciosa transformation system and represents an important step for further research into genetic manipulation of this fungal plant pathogen.