Agrobacterium tumefaciens-mediated genetic transformation of the phytopathogenic fungus Penicillium digitatum.
Agrobacterium tumefaciens-mediated transformation (ATMT) system was assessed for conducting insertional mutagenesis in Penicillium digitatum, a major fungal pathogen infecting post-harvest citrus fruits. A transformation efficiency of up to 60 transformants per 10(6) conidia was achieved by this system. The integration of the hph gene into the fungal genome was verified by polymerase chain reaction (PCR) amplification and sequencing. These transformants tested were also shown to be mitotically stable. Southern blot analysis of 14 randomly selected transformants showed that the hph gene was randomly integrated as single copy into the fungal genome of P. digitatum. Thus, we conclude that ATMT of P. digitatum could be used as an alternatively practical genetic tool for conducting insertional mutagenesis in P. digitatum to study functional genomics.
Wang
Ji-Ye
JY
Institute of Biotechnology, Zhejiang University, Hangzhou 310029, China.
Li
Hong-Ye
HY
D016960:Agrobacterium tumefaciens / Q000235:genetics*; D001483:Base Sequence; D002957:Citrus / Q000382:microbiology; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics; D004274:DNA, Recombinant / Q000235:genetics; D025141:Drug Resistance, Fungal / Q000235:genetics; D006921:Hygromycin B / Q000494:pharmacology; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D010407:Penicillium / Q000187:drug effects / Q000235:genetics* / Q000472:pathogenicity*; D010935:Plant Diseases / Q000382:microbiology; D010957:Plasmids / Q000235:genetics; D014170:Transformation, Genetic
D017931:DNA Primers; D004269:DNA, Bacterial; D004271:DNA, Fungal; D004274:DNA, Recombinant; C036483:T-DNA; D006921:Hygromycin B