1. Transformation of the entomopathogenic fungus Paecilomyces fumosoroseus with Agrobacterium tumefaciens.
- Author
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Lima, I. G. P., Duarte, R. T. D., Furlaneto, L., Baroni, C. H., Fungaro, M. H. P., and Furlaneto, M. C.
- Subjects
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AGROBACTERIUM tumefaciens , *PAECILOMYCES , *PATHOGENIC fungi , *ALEYRODIDAE , *ESCHERICHIA coli , *DNA - Abstract
Aims: 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. Methods and 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 105, 106 and 107 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. Significance and Impact of the Study: 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. [ABSTRACT FROM AUTHOR]
- Published
- 2006
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