Rapid detection of mutations and chromosomal rearrangements in T-DNA insertion mutants of Colletotrichum higginsianum using long-read whole-genome sequencing

Random mutagenesis is a useful tool for identifying genes of interest. Here, we used long-read whole-genome sequencing analysis to identify T-DNA insertion sites in pathogenicity mutants of Colletotrichum higginsianum, a fungus that causes anthracnose on Brassica plants. We identified mutated regions by aligning the sequence reads containing T-DNA to the C. higginsianum reference genome and also detected unexpectedly large chromosomal rearrangements that appear to have been caused by T-DNA transformation. These results demonstrate that long-read whole-genome sequencing is effective for identifying mutated regions in the C. higginsianum mutants.