Construction and high-throughput phenotypic screening of Zymoseptoria tritici over-expression strains

•We generate a pilot library of 32 Z. tritici over-expression strains.•We develop a high throughput technique for in vitro phenotypic screening.•This protocol can probe both hyphal and budding Z. tritici morphologies.•A putative transcription factor impedes hyphal biosynthesis when over-expressed.•Genome wide functional analysis will enable discovery of novel virulence attributes.