Agrobacterium tumefaciens-mediated transformation as an efficient tool for insertional mutagenesis of Kabatiella zeae

Agrobacterium tumefaciens-mediated transformation (ATMT) has been widely used in filamentous fungi. In this study, an efficient Agrobacterium tumefaciens-mediated transformation approach was developed for the plant pathogenic fungus, Kabatiella zeae, the causative pathogen of eyespot in maize. Five parameters were selected to optimize efficiencies of transformation. A. tumefaciens concentration, conidia concentration of K. zeae and mixing ratio of A. tumefaciens and K. zeae were found to exert a significant influence on all parameters. Transformants emitted green fluorescence under fluorescence microscopy. The presence of mitotically stable hygromycin resistance gene (hph) integration in the genome was confirmed by PCR. Up to 148 transformants per 107 conidia could be obtained under optimal conditions. In this way, ATMT approach is an efficient tool for insertional mutagenesis of K. zeae.