Miao Zhengang, Wang Guangyuan, Shen Heng, Wang Xue, Gabriel Dean W, Liang Wenxing
College of Plant Health and Medicine, Engineering Research Center for Precision Pest Management for Fruits and Vegetables of Qingdao, Shandong Engineering Research Center for Environment-Friendly Agricultural Pest Management, Shandong Province Key Laboratory of Applied Mycology, Qingdao Agricultural University, Qingdao, China.
College of Life Sciences, Shandong Province Key Laboratory of Applied Mycology, Qingdao Agricultural University, Qingdao, China.
Front Microbiol. 2022 Jun 30;13:925868. doi: 10.3389/fmicb.2022.925868. eCollection 2022.
DNA adenine N-methylation (6mA) plays a critical role in various biological functions, but its occurrence and functions in filamentous plant pathogens are largely unexplored. is an important pathogenic fungus worldwide. A systematic analysis of 6mA in was performed in this study, revealing that 6mA is widely distributed in the genome of this fungus. The 2 kb regions flanking many genes, particularly the upstream promoter regions, were susceptible to methylation. The role of BcMettl4, a 6mA methyltransferase, in the virulence of was investigated. disruption and point mutations of its catalytic motif "DPPW" both resulted in significant 6mA reduction in the genomic DNA and in reduced virulence of . RNA-Seq analysis revealed a total of 13 downregulated genes in the disruption mutant ΔBcMettl4 in which methylation occurred at the promoter sites. These were involved in oxidoreduction, secretory pathways, autophagy and carbohydrate metabolism. Two of these genes, and , were independently disrupted. Knockout of led to reduced sclerotium formation, while disruption of resulted in dramatically decreased conidium formation and pathogenicity. These observations indicated that 6mA provides potential epigenetic markers in and that BcMettl4 regulates virulence in this important plant pathogen.
DNA腺嘌呤N-甲基化（6mA）在多种生物学功能中起关键作用，但其在丝状植物病原体中的发生情况和功能在很大程度上尚未得到探索。是全球一种重要的致病真菌。本研究对中的6mA进行了系统分析，发现6mA广泛分布于该真菌的基因组中。许多基因两侧的2kb区域，尤其是上游启动子区域，易发生甲基化。研究了6mA甲基转移酶BcMettl4在致病力中的作用。其催化基序“DPPW”的破坏和点突变均导致基因组DNA中6mA显著减少，并降低了的致病力。RNA测序分析显示，在破坏突变体ΔBcMettl4中共有13个下调基因，其启动子位点发生了甲基化。这些基因参与氧化还原、分泌、自噬和碳水化合物代谢。其中两个基因，和，被独立破坏。敲除导致菌核形成减少，而破坏导致分生孢子形成和致病性显著降低。这些观察结果表明，6mA在中提供了潜在的表观遗传标记，并且BcMettl4调节这种重要植物病原体的致病力。
