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.
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