Development of a transformation system for Crinipellis perniciosa, the causal agent of witches' broom in cocoa plants

• Autor(a) principal: Lima, Juliana Oliveira
• Data de Publicação: 2002
• Outros Autores: Santos, Jildete Karla dos, Pereira, Jorge Fernando, Resende, Mário Lúcio Vilela de, Araújo, Elza Fernandes de, Queiroz, Marisa Vieira de
• Tipo de documento: Artigo
• Idioma: eng
• Título da fonte: LOCUS Repositório Institucional da UFV
• Texto Completo: https://link.springer.com/article/10.1007/s00294-002-0358-z; http://www.locus.ufv.br/handle/123456789/18912
• topic: Transformation; Hygromycin resistance; Witches' broom

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.