Cochliobolus heterostrophus is a plant pathogenic fungus of southern corn leaf blight, which has been regarded as a model necrotrophic plant pathogen. Many methods have been developed to knock out targeted genes in C. heterostrophus, of which the most widely-used one is protoplast-mediated transformation. However, there are several problems of this method associated with protoplast preparation, DNA product, time consumption, or high cost. In this study, a highly efficient target gene deletion approach in C. heterostrophus was established and optimized, based on Agrobacterium tumefaciens-mediated transformation (ATMT); the transformation efficiency of this approach was 85–88 transformants per 10⁵ conidia, and the homologous recombination efficiency was approximately 68.3%. Furthermore, six gene knockout mutants of C. heterostrophus were obtained using this ATMT method. The phenotypes of this fungus altered in the mutant strains, and the virulence of the mutants significantly reduced compared to of the wild type strain. Taken together, this ATMT system established in this study can be used as a genetic manipulation tool for C. heterostrophus, to better understand the functions of genes and its relation to virulence.

异旋旋孢菌是南方玉米叶枯病的植物病原真菌，被认为是坏死营养型植物病原菌的模型。已经开发了许多方法来敲除异旋梭菌中的靶基因，其中最广泛使用的一种是原生质体介导的转化。然而，该方法存在原生质体制备、DNA产物、耗时或成本高等问题。本研究基于根癌农杆菌介导的转化（ATMT），建立并优化了异旋梭菌的高效靶基因删除方法；该方法的转化效率为每10 ^{5 个}分生孢子85-88个转化子，同源重组效率约为68.3%。此外，利用该ATMT方法还获得了C.异旋菌的6个基因敲除突变体。该真菌的表型在突变株中发生了改变，与野生型菌株相比，突变株的毒力显着降低。综上所述，本研究建立的ATMT系统可以作为异旋梭菌的遗传操作工具，以更好地了解基因的功能及其与毒力的关系。