Zhai Ziqi, Zhang Mengwei, Yin Ruya, Zhao Siji, Shen Zhen, Yang Yonglin, Zhang Xuan, Wang Jianing, Qin Yifei, Xu Dan, Zhou Ligang, Lai Daowan
State Key Laboratory of Agricultural and Forestry Biosecurity, College of Plant Protection, China Agricultural University, Beijing, 100193, China.
Department of Plant Pathology and MOA Key Lab of Pest Monitoring and Green Management, College of Plant Protection, China Agricultural University, Beijing, 100193, China.
World J Microbiol Biotechnol. 2025 Mar 12;41(3):103. doi: 10.1007/s11274-025-04313-x.
Edenia gomezpompae, an endophytic fungus derived from plants, produced a diverse array of preussomerins, a type of spirobisnaphthalenes featuring two spiroketal groups, which exhibited significant antibacterial, antifungal, and cytotoxic activities. Structurally, the biosynthesis of preussomerins might be related to the biosynthesis of 1,8-dihydroxynaphthalene (DHN), a precursor of DHN-melanin. However, the absence of efficient gene-editing tools for E. gomezpompae has hindered the biosynthetic study of preussomerins. In this study, we developed a CRISPR/Cas9-based gene editing system for E. gomezpompae SV2 that was isolated from the stem of Setaria viridis, by utilizing the endogenous U6 snRNA promoter to drive sgRNA expression. Using this system, we successfully disrupted the polyketide synthase (PKS)-encoding gene, Egpks, a putative 1,3,6,8-tetrahydroxynaphthalene synthase gene involved in the biosynthesis of DHN-melanin, with an editing efficiency up to 92% and a knockout efficiency of 71% when employing the U6 snRNA-3 promoter. Furthermore, the disrupted mutant (∆Egpks) displayed white hyphae and lost the ability to produce preussomerins. These results provided a foundational tool for genetic manipulation in E. gomezpompae and revealed the role of EgPKS in the biosynthesis of preussomerin-type spirobisnaphthalenes.
戈麦斯蓬帕伊艾氏菌是一种源自植物的内生真菌，它能产生多种前乌索菌素，这是一类具有两个螺环缩酮基团的螺二萘类化合物，具有显著的抗菌、抗真菌和细胞毒性活性。在结构上，前乌索菌素的生物合成可能与1,8 - 二羟基萘（DHN）的生物合成有关，DHN是DHN - 黑色素的前体。然而，缺乏针对戈麦斯蓬帕伊艾氏菌的高效基因编辑工具阻碍了前乌索菌素的生物合成研究。在本研究中，我们利用内源性U6 snRNA启动子驱动sgRNA表达，为从绿色狗尾草茎中分离出的戈麦斯蓬帕伊艾氏菌SV2开发了一种基于CRISPR/Cas9的基因编辑系统。利用该系统，我们成功破坏了编码聚酮合酶（PKS）的基因Egpks，这是一个推测参与DHN - 黑色素生物合成的1,3,6,8 - 四羟基萘合酶基因，当使用U6 snRNA - 3启动子时，编辑效率高达92%，敲除效率为71%。此外，破坏后的突变体（∆Egpks）呈现白色菌丝，失去了产生前乌索菌素的能力。这些结果为戈麦斯蓬帕伊艾氏菌的遗传操作提供了一个基础工具，并揭示了EgPKS在前乌索菌素型螺二萘生物合成中的作用。
