Raw data_(Submission ID 247694791)

Background: Reactive oxygen species (ROS), synthesized by the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase (Nox) complex, are vital molecules in biological cells, influencing various physiological processes such as fungal growth, development, and virulence. Beauveria bassiana, an entomopathogenic fungus, is a promising biopesticide for agricultural, forestry, and urban pest control. This study focuses on the characterization of NADPH oxidases (Noxs) in B. bassiana. Materials and Methods: Gene expression profiles of Noxs in B. bassiana (BbNoxs) were analyzed using RT-qPCR. Knockout strains of single BbNoxA, BbNoxB, BbNoxR, and double BbNoxA and BbNoxB were constructed via homologous recombination, and their phenotypic characteristics were examined. Fungal virulence was evaluated using Galleria mellonella larvae, and infection structures formation and penetration ability were assessed on cicada wings. ROS production and actin assembly during fungal growth and infection were detected using staining and marker methods. Results: Expression analysis revealed significant upregulation of BbNoxs during fungal growth and infection. Compared to the wild-type strain, single knockouts (ΔBbNoxA/B/R) and double knockout (ΔBbNoxAB) of BbNoxs exhibited reduced conidial yields, accelerated conidial germination rates. Deletion of BbNoxB or BbNoxR decreased fungal virulence compared to the WT strain in topical inoculation experiments. Additionally, loss of BbNoxB or BbNoxR impaired infection structures formation, penetration ability, ROS production, and actin aggregation during fungal infection. Conclusion: BbNoxs are crucial for fungal growth, development, and virulence in B. bassiana, playing essential roles in infection structures formation, penetration, ROS production, and actin assembly. Understanding their functions provides insights into B. bassiana’s pathogenic mechanisms.背景：活性氧（Reactive oxygen species, ROS）是由烟酰胺腺嘌呤二核苷酸磷酸（nicotinamide adenine dinucleotide phosphate, NADPH）氧化酶（Nox）复合物合成的生物细胞内关键分子，参与调控真菌生长、发育及毒力等诸多生理过程。球孢白僵菌（Beauveria bassiana）作为一类虫生真菌（entomopathogenic fungus），是用于农业、林业及城市害虫防治的极具潜力的生物农药。本研究围绕球孢白僵菌中的NADPH氧化酶（Noxs）展开功能表征工作。 材料与方法：采用实时定量聚合酶链反应（RT-qPCR）分析球孢白僵菌中Noxs（命名为BbNoxs）的基因表达谱。通过同源重组（homologous recombination）技术构建单基因敲除菌株ΔBbNoxA、ΔBbNoxB、ΔBbNoxR以及双基因敲除菌株ΔBbNoxAΔBbNoxB，并对其表型特征进行检测。以大蜡螟（Galleria mellonella）幼虫为试虫评估菌株毒力，以蝉翼为载体检测菌株的侵染结构形成能力与穿透能力。采用染色与标记技术检测真菌生长及侵染过程中的ROS产生与肌动蛋白（actin）组装情况。 结果：表达分析结果显示，BbNoxs在真菌生长及侵染阶段均显著上调表达。与野生型菌株相比，单基因敲除菌株（ΔBbNoxA/B/R）及双基因敲除菌株ΔBbNoxAB的分生孢子产量下降，分生孢子萌发速率加快。在体表接种实验中，敲除BbNoxB或BbNoxR会导致菌株毒力较野生型菌株显著下降。此外，敲除BbNoxB或BbNoxR会损害真菌侵染过程中的侵染结构形成能力、穿透能力、ROS产生及肌动蛋白聚集过程。 结论：BbNoxs对球孢白僵菌的真菌生长、发育及毒力至关重要，在侵染结构形成、穿透能力、ROS产生及肌动蛋白组装过程中发挥核心作用。阐明其功能可为解析球孢白僵菌的致病机制提供理论依据。