Transformation of the entomopathogenic fungus Paecilomyces fumosoroseus with Agrobacterium tumefaciens.
OBJECTIVE
To test the suitability of the Agrobacterium tumefaciens-mediated transformation (AMT) method with Paecilomyces fumosoroseus, a fungal pathogen that causes diseases in a wide range of insects including whiteflies.


RESULTS
Conidia of P. fumosoroseus were successfully transformed to hygromycin B resistance using the hph gene of Escherichia coli as the selectable marker. Transformation frequencies were 58.3 +/- 18.5, 98.3 +/- 24.8 and 169.7 +/- 35.5 (+/-SEM) transformants per 10(5), 10(6) and 10(7) target conidia respectively. After confirmation by PCR, transformants were subjected to Southern analysis, and the results revealed that 45% (four of nine) of the transformants contained single-copy integration of the T-DNA.


CONCLUSIONS
In our AMT system, we efficiently transformed conidia of P. fumosoroseus. The employment of this method circumvents time-consuming protoplast preparation and allows the isolation of transformants containing single-copy integration of the T-DNA.


CONCLUSIONS
Considering the efficiency of Ag. tumefaciens-mediated transformation, this method represents a useful tool for insertional mutagenesis to characterize genes that are important for the pathogenicity of P. fumosoroseus.
Lima
I G P
IG
Centro de Ciências Biológicas, Universidade Estadual de Londrina, Londrina-PR, Brazil.
Duarte
R T D
RT
Furlaneto
L
L
Baroni
C H
CH
Fungaro
M H P
MH
Furlaneto
M C
MC
D016960:Agrobacterium tumefaciens / Q000235:genetics*; D000818:Animals; D024881:Drug Resistance, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial; D006921:Hygromycin B / Q000494:pharmacology; D007313:Insecta / Q000382:microbiology*; D010143:Paecilomyces / Q000235:genetics* / Q000472:pathogenicity*; D014170:Transformation, Genetic
D006921:Hygromycin B