Chinese Academy of Agricultural Sciences/Key Laboratory of Agro-products Quality and Safety Control in Storage and Transport Process, Ministry of Agriculture and Rural Affairs, Institute of Food Science and Technology, Beijing 100193, P. R. China.
J Agric Food Chem. 2024 Jun 26;72(25):14229-14240. doi: 10.1021/acs.jafc.4c01914. Epub 2024 May 26.
() is a globally recognized and highly impactful fungal pathogen of maize, causing yield losses and producing harmful mycotoxins that pose a threat to human and animal health. However, the genetic tools available for studying this crucial fungus are currently limited in comparison to other important fungal pathogens. To address this, an efficient CRISPR/Cas9 genome editing system based on an autonomously replicating plasmid with an AMA1 sequence was established in this study. First, gene disruption of and via nonhomologous end-joining (NHEJ) pathway was successfully achieved, with efficiency ranging from 66 to 100%. Second, precise gene deletions were achieved with remarkable efficiency using a dual sgRNA expression strategy. Third, the developed genome editing system can be applied to generate designer chromosomes in , as evidenced by the deletion of a crucial 38 kb fragment required for fumonisin biosynthesis. Fourth, the recycling system has been established and successfully applied in . Lastly, the developed Δ and Δ mutants can serve as biocontrol agents to reduce the fumonisin B (FB) contamination produced by the toxigenic strain. Taken together, these significant advancements in genetic manipulation and biocontrol strategies provide valuable tools for studying and mitigating the impact of on maize crops.
（）是一种全球公认的、具有高度影响力的玉米真菌病原体，它会导致产量损失，并产生有害的真菌毒素，从而对人类和动物健康构成威胁。然而，与其他重要的真菌病原体相比，目前用于研究这种关键真菌的遗传工具仍然有限。为了解决这个问题，本研究建立了一种基于含有 AMA1 序列的自主复制质粒的高效 CRISPR/Cas9 基因组编辑系统。首先，通过非同源末端连接（NHEJ）途径成功实现了和的基因敲除，效率范围为 66%至 100%。其次，利用双 sgRNA 表达策略实现了精确的基因缺失，效率显著。第三，所开发的基因组编辑系统可用于在中生成设计染色体，这可从缺失一个对伏马菌素生物合成至关重要的 38kb 片段得到证明。第四，已经建立了并成功应用于。最后，开发的Δ和Δ突变体可以作为生物防治剂，减少由产毒菌株产生的伏马菌素 B（FB）污染。总之，这些遗传操作和生物防治策略的重大进展为研究和减轻对玉米作物的影响提供了有价值的工具。
