Hao Luyao, Liu Hongyou, Zhao Fengmiao, Ma Yuan, Li Zhengyi, Wang Rui
College of Veterinary Medicine, Inner Mongolia Agricultural University, Hohhot, Inner Mongolia, People's Republic of China; Key Laboratory of Clinical Diagnosis and Treatment of Animal Diseases, Ministry of Agriculture, National Animal Medicine Experimental Teaching Center, People's Republic of China.
College of Veterinary Medicine, Inner Mongolia Agricultural University, Hohhot, Inner Mongolia, People's Republic of China; Key Laboratory of Clinical Diagnosis and Treatment of Animal Diseases, Ministry of Agriculture, National Animal Medicine Experimental Teaching Center, People's Republic of China.
Vet Parasitol. 2025 Jul;337:110463. doi: 10.1016/j.vetpar.2025.110463. Epub 2025 Apr 23.
Traditional parasite management has long been plagued by issues such as drug resistance and environmental pollution. Biological control using Pochonia chlamydosporia has emerged as a sustainable alternative, yet the underlying infection mechanisms remain elusive. This study aimed to comprehensively elucidate these mechanisms, with a particular focus on the role of gene editing. Employing optical, SEM, and TEM microscopy, we observed that P. chlamydosporia infects Fasciola hepatica eggs in three distinct stages. TEM analysis first visualized unique infection pegs crucial for initial eggshell penetration. Using 4D-DIA mass spectrometry, proteomics identified 208 differentially expressed proteins between normal and nematode - egg - induced mycelium, of which 93 were downregulated and 115 were upregulated. Through comprehensive protein sequencing and subsequent bioinformatics analyses, we successfully identified a key gene, designated p1. To understand p1's function, we used RNA interference (RNAi) and overexpression. We constructed pSilent-1-p1 (RNAi vector) and pBARGPE1-p1 (overexpression vector). The overexpression strain pBARGPE-p1 demonstrated a remarkable increase in serine protease activity (0.63 U/mL), indicating an enhanced ability to degrade host tissues. Conversely, the deletion strain pSilent-1-p1 had lower activity, indicating the crucial role of the p1 gene in protease production. When assessing the infection efficiency against three types of nematode eggs, both the overexpressed and silenced strains exhibited a downward trend. The silenced strain had a significantly reduced infection rate, with an average of 45.22 %, highlighting the importance of the p1 gene in the fungus's parasitic ability. Notably, no significant differences were observed among strains with respect to spore concentration, mycelial biomass, and growth rate. These findings, centered on the p1 gene provide comprehensive insights into the biological control mechanisms of P. chlamydosporia, establishing a solid theoretical foundation for the development of more efficient and environmentally friendly parasite management strategies.
传统的寄生虫管理长期以来一直受到耐药性和环境污染等问题的困扰。利用厚垣普可尼亚菌进行生物防治已成为一种可持续的替代方法，但其潜在的感染机制仍不清楚。本研究旨在全面阐明这些机制，特别关注基因编辑的作用。通过光学显微镜、扫描电子显微镜和透射电子显微镜，我们观察到厚垣普可尼亚菌感染肝片吸虫卵分为三个不同阶段。透射电镜分析首次观察到了对初始卵壳穿透至关重要的独特感染钉。利用4D-DIA质谱技术，蛋白质组学鉴定出正常菌丝体和线虫卵诱导的菌丝体之间有208种差异表达蛋白，其中93种下调，115种上调。通过全面的蛋白质测序和后续的生物信息学分析，我们成功鉴定出一个关键基因，命名为p1。为了解p1的功能，我们使用了RNA干扰（RNAi）和过表达技术。我们构建了pSilent-1-p1（RNAi载体）和pBARGPE1-p1（过表达载体）。过表达菌株pBARGPE-p1的丝氨酸蛋白酶活性显著增加（0.63 U/mL），表明其降解宿主组织的能力增强。相反，缺失菌株pSilent-1-p1的活性较低，表明p1基因在蛋白酶产生中起关键作用。在评估对三种线虫卵的感染效率时，过表达菌株和沉默菌株均呈现下降趋势。沉默菌株的感染率显著降低，平均为45.22%，突出了p1基因在该真菌寄生能力中的重要性。值得注意的是，在孢子浓度、菌丝体生物量和生长速率方面，各菌株之间未观察到显著差异。这些以p1基因为中心的研究结果为厚垣普可尼亚菌的生物防治机制提供了全面的见解，为开发更高效、更环保的寄生虫管理策略奠定了坚实的理论基础。
