Development of a transformation system for Crinipellis perniciosa, the causal agent of witches' broom in cocoa plants.
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.
Lima
Juliana Oliveira
JO
Departamento de Microbiologia/BIOAGRO, Universidade Federal de Viçosa, 36.570-000 Viçosa, Minas Gerais, Brazil.
dos Santos
Jildete Karla
JK
Pereira
Jorge Fernando
JF
de Resende
Mário Lúcio Vilela
ML
de Araújo
Elza Fernandes
EF
de Queiroz
Marisa Vieira
MV
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
D000900:Anti-Bacterial Agents; D002934:Cinnamates; D006921:Hygromycin B; C026273:hygromycin A