High frequency gene conversion among benomyl resistant transformants in the entomopathogenic fungus Metarhizium anisopliae.
Three different methods, (i) PEG, (ii) electroporation and (iii) biolistic, were employed to transform Metarhizium anisopliae using benomyl resistance as a selectable marker. Transformation frequencies and mitotic stability varied for each method, from 0.8 to 6.9 transformants micrograms-1 of DNA and 46%, respectively, by the PEG method; 1.3 to 1.8 transformants micrograms-1 of DNA and 67% by electroporation; and 32 to 201 transformants micrograms-1 of DNA and 90% by biolistic. We demonstrate by PCR that 60% of the transformants were generated by gene conversion.
Bogo
M R
MR
Departamento de Biotecnologia, Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil.
Vainstein
M H
MH
Aragão
F J
FJ
Rech
E
E
Schrank
A
A
D000818:Animals; D001483:Base Sequence; D001542:Benomyl / Q000494:pharmacology*; D017931:DNA Primers / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics; D004352:Drug Resistance, Microbial / Q000235:genetics*; D018274:Electroporation; D005659:Fungicides, Industrial / Q000494:pharmacology*; D005785:Gene Conversion; D005819:Genetic Markers; D003904:Mitosporic Fungi / Q000187:drug effects* / Q000235:genetics* / Q000472:pathogenicity; D008969:Molecular Sequence Data; D010572:Pest Control, Biological; D011092:Polyethylene Glycols; D014170:Transformation, Genetic
D017931:DNA Primers; D004271:DNA, Fungal; D005659:Fungicides, Industrial; D005819:Genetic Markers; D011092:Polyethylene Glycols; D001542:Benomyl