title: Development of a transformation system for Crinipellis perniciosa, the causal agent of witches' broom in cocoa plants.
abstract: 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/CaCl(2) treatment. Transformation frequencies of 1.6-2.5 transformants/microg 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.
authors.affiliation: Departamento de Microbiologia/BIOAGRO, Universidade Federal de Viçosa, 36.570-000 Viçosa, Minas Gerais, Brazil.
authors.forename: Juliana Oliveira
authors.initials: JO
authors.lastname: Lima
authors.forename: Jildete Karla
authors.initials: JK
authors.lastname: dos Santos
authors.forename: Jorge Fernando
authors.initials: JF
authors.lastname: Pereira
authors.forename: Mário Lúcio Vilela
authors.initials: ML
authors.lastname: de Resende
authors.forename: Elza Fernandes
authors.initials: EF
authors.lastname: de Araújo
authors.forename: Marisa Vieira
authors.initials: MV
authors.lastname: de Queiroz
mesh_terms: D000900:Anti-Bacterial Agents / Q000378:metabolism; D001203:Ascomycota / Q000235:genetics*; D002099:Cacao / Q000382:microbiology; D015825:Chromosomes, Fungal; D002934:Cinnamates / Q000378:metabolism; D025141:Drug Resistance, Fungal / Q000235:genetics*; D005800:Genes, Fungal; D006921:Hygromycin B / Q000031:analogs & derivatives* / Q000378:metabolism; D014162:Transfection
chemical_list: D000900:Anti-Bacterial Agents; D002934:Cinnamates; D006921:Hygromycin B; C026273:hygromycin A