AGROBACTERIUM-MEDIATED TRANSFORMATION OF THE PLANT PATHOGENIC FUNGUS VERTICILLIUM ALBO-ATRUM

Abstract

We report the successful Agrobacterium tumefaciensmediated transformation of the important plant pathogen Verticillium albo-atrum. V. albo-atrum was demonstrated to be hygromycin B sensitive and consequently transformations were based on integration of either the hygromycin B resistance gene (hph) or hph in addition to the fluorescence gene DsRed. The effect of various parameters on transformation efficiency, including spore concentration, acetosyringone concentration, selection media and co-cultivation time were investigated. Transformants were analysed by PCR and Southern analysis, and were found to contain randomly integrated T-DNA that typically inserted as one or two copies. The dual marker system afforded by pCAMDsRed provides an additional and fast method of transformant verification. Development of a successful transformation system for V. albo-atrum should facilitate further molecular studies of this important plant pathogen.