Raw data_(Submission ID 247694791)

<b>Background:</b> 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. <i>Beauveria bassiana</i>, 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 <i>B. bassiana</i>.<b>Materials and Methods:</b> Gene expression profiles of <i>Noxs</i> in <i>B. bassiana</i> (<i>BbNoxs</i>) were analyzed using RT-qPCR. Knockout strains of single <i>BbNoxA</i>, <i>BbNoxB</i>, <i>BbNoxR</i>, and double <i>BbNoxA</i> and <i>BbNoxB</i> were constructed via homologous recombination, and their phenotypic characteristics were examined. Fungal virulence was evaluated using <i>Galleria mellonella</i> 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.<b>Results:</b> Expression analysis revealed significant upregulation of <i>BbNoxs</i> during fungal growth and infection. Compared to the wild-type strain, single knockouts (Δ<i>BbNoxA/B/R</i>) and double knockout (Δ<i>BbNoxAB</i>) of <i>BbNoxs</i> exhibited reduced conidial yields, accelerated conidial germination rates. Deletion of <i>BbNoxB</i> or <i>BbNoxR</i> 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.<b>Conclusion:</b> BbNoxs are crucial for fungal growth, development, and virulence in <i>B. bassiana</i>, playing essential roles in infection structures formation, penetration, ROS production, and actin assembly. Understanding their functions provides insights into <i>B. bassiana</i>’s pathogenic mechanisms.