Liu Qianqian, Jiang Kexin, Duan Shipeng, Zhao Na, Shen Yanmei, Zhu Lirong, Zhang Ke-Qin, Yang Jinkui
State Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan, Key Laboratory for Southwest Microbial Diversity of the Ministry of Education, and School of Life Science, Yunnan University, Kunming 650032, China.
State Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan, Key Laboratory for Southwest Microbial Diversity of the Ministry of Education, and School of Life Science, Yunnan University, Kunming 650032, China.
J Adv Res. 2025 Feb;68:1-15. doi: 10.1016/j.jare.2024.02.002. Epub 2024 Feb 7.
Arthrobotrys oligospora has been utilized as a model strain to study the interaction between fungi and nematodes owing to its ability to capture nematodes by developing specialized traps. A previous study showed that high-osmolarity glycerol (Hog1) signaling regulates the osmoregulation and nematocidal activity of A. oligospora. However, the function of downstream transcription factors of the Hog1 signaling in the nematode-trapping (NT) fungi remains unclear.
This study aimed to investigate the functions and potential regulatory network of AoMsn2, a downstream transcription factor of the Hog1 signaling pathway in A. oligospora.
The function of AoMsn2 was characterized using targeted gene deletion, phenotypic experiments, real-time quantitative PCR, RNA sequencing, untargeted metabolomics, and yeast two-hybrid analysis.
Loss of Aomsn2 significantly enlarged and swollen the hyphae, with an increase in septa and a significant decrease in nuclei. In particular, spore yield, spore germination rate, traps, and nematode predation efficiency were remarkably decreased in the mutants. Phenotypic and transcriptomic analyses revealed that AoMsn2 is essential for fatty acid metabolism and autophagic pathways. Additionally, untargeted metabolomic analysis identified an important function of AoMsn2 in the modulation of secondary metabolites. Furtherly, we analyzed the protein interaction network of AoMsn2 based on the Kyoto Encyclopedia of Genes and Genomes pathway map and the online website STRING. Finally, Hog1 and six putative targeted proteins of AoMsn2 were identified by Y2H analysis.
Our study reveals that AoMsn2 plays crucial roles in the growth, conidiation, trap development, fatty acid metabolism, and secondary metabolism, as well as establishes a broad basis for understanding the regulatory mechanisms of trap morphogenesis and environmental adaptation in NT fungi.
少孢节丛孢菌因其能够通过形成特殊陷阱捕获线虫的能力，被用作研究真菌与线虫相互作用的模式菌株。先前的一项研究表明，高渗甘油（Hog1）信号通路调节少孢节丛孢菌的渗透压调节和杀线虫活性。然而，Hog1信号通路的下游转录因子在捕食线虫（NT）真菌中的功能仍不清楚。
本研究旨在探究少孢节丛孢菌中Hog1信号通路的下游转录因子AoMsn2的功能及其潜在调控网络。
通过靶向基因缺失、表型实验、实时定量PCR、RNA测序、非靶向代谢组学和酵母双杂交分析对AoMsn2的功能进行表征。
Aomsn2缺失显著使菌丝增大、肿胀，隔膜增加，细胞核显著减少。特别是，突变体中的孢子产量、孢子萌发率、陷阱和线虫捕食效率显著降低。表型和转录组分析表明，AoMsn2对脂肪酸代谢和自噬途径至关重要。此外，非靶向代谢组分析确定了AoMsn2在次生代谢产物调节中的重要功能。进一步地，我们基于京都基因与基因组百科全书通路图和在线网站STRING分析了AoMsn2的蛋白质相互作用网络。最后，通过酵母双杂交分析鉴定了Hog1和AoMsn2的六个假定靶向蛋白。
我们的研究表明，AoMsn2在生长、产孢、陷阱发育、脂肪酸代谢和次生代谢中起关键作用，并为理解NT真菌中陷阱形态发生和环境适应的调控机制奠定了广泛基础。
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