An improved Agrobacterium-mediated transformation system for the functional genetic analysis of Penicillium marneffei

We have developed an improved Agrobacterium-mediated transformation (AMT) system for the functional genetic analysis of Penicillium marneffei, a thermally dimorphic, human pathogenic fungus. Our AMT protocol included the use of conidia or pre-germinated conidia of P. marneffei as the host recipient for T-DNA from Agrobacterium tumefaciens and co-cultivation at 28 degrees C for 36 hours. Bleomycin-resistant transformants were selected as yeast-like colonies following incubation at 37 degrees C. The efficiency of transformation was approximately 123 +/- 3.27 and 239 +/- 13.12 transformants per plate when using 5 x 10