Shao Lei, Liu Zuoyi, Liu Yongxiang, Tan Yumei
College of Pharmacy, Guiyang Healthcare Vocational University, Guiyang, Guizhou, China.
Guizhou Provincial Engineering Research Center of Medical Resourceful Healthcare Products, Guiyang Healthcare Vocational University, Guiyang, Guizhou, China.
J Basic Microbiol. 2025 Jan;65(1):e2400475. doi: 10.1002/jobm.202400475. Epub 2024 Oct 7.
Aspergillus cristatus is a dominant fungus formed during the "flowering" process of Fuzhuan brick tea. Previous research has established that the sporulation of Aspergillus nidulans, a model organism of filamentous fungi, is regulated by light. However, the sporulation of A. cristatus is dependent on osmotic stress. In a previous study, we used pull-down and mass spectrometry to identify proteins that interacted with AcHog1 in A. cristatus when cultured under different conditions of osmotic stress. In the present study, we analyzed the proteins we identified previously to investigate their functional role. The AA1E3BER4 protein was located downstream of Hog1 in the HOG branch pathway and was identified that was regulated by AcHog1. Furthermore, yeast two-hybrid analysis showed that AA1E3BER4 interacted with AcHog1. In addition, we knocked out and complemented the Acsko1 gene encoding the AA1E3BER4 protein. We found that the number of sexual and asexual spores were downregulated by 3.81- and 4.57-fold, respectively, in the ΔAcsko1 strain. The sensitivity of the ΔAcsko1 strain to sorbitol and sucrose, as regulators of osmotic stress, increased, and the sensitivity to high sucrose was higher than that of sorbitol. Acsko1 also regulated the response of A. cristatus to oxidative stress, Congo red, and SDS (sodium dodecyl sulfate). In addition, the deletion of Acsko1 significantly increased the pigment of the ΔAcsko1 strain. This is the first study to report the role of the sko1 gene in oxidative stress, stress-induced damage to the cell wall, and pigment in Aspergillus cristatus.
冠突曲霉是茯砖茶“发花”过程中形成的优势真菌。先前的研究表明，丝状真菌的模式生物构巢曲霉的孢子形成受光调节。然而，冠突曲霉的孢子形成依赖于渗透胁迫。在之前的一项研究中，我们使用下拉法和质谱法鉴定了在不同渗透胁迫条件下培养的冠突曲霉中与AcHog1相互作用的蛋白质。在本研究中，我们分析了之前鉴定的蛋白质以研究它们的功能作用。AA1E3BER4蛋白位于HOG分支途径中Hog1的下游，并被鉴定受AcHog1调控。此外，酵母双杂交分析表明AA1E3BER4与AcHog1相互作用。另外，我们敲除并互补了编码AA1E3BER4蛋白的Acsko1基因。我们发现，在ΔAcsko1菌株中，有性和无性孢子的数量分别下调了3.81倍和4.57倍。作为渗透胁迫调节剂，ΔAcsko1菌株对山梨醇和蔗糖的敏感性增加，并且对高蔗糖的敏感性高于山梨醇。Acsko1还调节冠突曲霉对氧化应激、刚果红和十二烷基硫酸钠（SDS）的反应。此外，Acsko1的缺失显著增加了ΔAcsko1菌株的色素含量。这是首次报道sko1基因在冠突曲霉的氧化应激、应激诱导的细胞壁损伤和色素形成中的作用的研究。
