捕获线虫的真菌对几丁质反应后的蛋白质组学研究。

IF 1.3 3区 农林科学 Q2 VETERINARY SCIENCES
Journal of Veterinary Research Pub Date : 2025-02-25 eCollection Date: 2025-03-01 DOI:10.2478/jvetres-2025-0005
Jiahua Zhang, Lixiang Wei, Huimei Zhang, Xixi Ma, Yansen Sun, Ruobing Li, Chengzhi Zhang, Xuepeng Cai, Jun Qiao, Qingling Meng
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引用次数: 0

摘要

诱捕线虫真菌(NTFs)在捕食过程中可产生多种几丁质酶降解线虫体壁和蛋壳几丁质。然而,它们表达几丁质酶的调控机制尚不清楚。本研究的主要目的是阐明A. oligospora(一种NTF)对几丁质反应的差异蛋白谱。材料与方法:加入胶体几丁质诱导少孢霉培养,倒置显微镜下观察诱导前后的表型差异。采用液相色谱-串联质谱法筛选菌丝诱导前后的差异蛋白。将差异表达的几丁质酶在毕赤酵母中表达,并将重组酶与秀丽隐杆线虫及其卵悬液孵育,探讨其生物活性。结果:寡孢霉与甲壳素相互作用后菌丝生长明显加快。在对照组(AO-c)和实验组(AO-e)之间共鉴定出1124个差异表达蛋白(DEPs),其中上调183个,下调941个。基因本体分析表明,DEPs参与多种代谢过程,具有催化和结合功能。京都基因和基因组百科分析将这些蛋白质主要与葡萄糖代谢相关的信号通路联系起来。三种几丁质酶在dep中显著调节。此外,酶活性测定表明,其中一个有效地降解秀丽隐杆线虫及其卵。结论:寡孢单胞菌对甲壳素的响应可显著改变其蛋白表达谱,从而促进其糖代谢和菌丝发育。本研究为少孢单胞虫捕食线虫的调控机制提供了新的认识。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Proteomic insights into nematode-trapping fungi Arthrobotrys oligospora after their response to chitin.

Introduction: Nematode-trapping fungi (NTFs) can produce various chitinases to degrade nematode body wall and eggshell chitin during predation. However, the regulatory mechanisms of their expression of chitinases still remain unclear. The primary objective of this study was to elucidate the differential protein profile of A. oligospora, an NTF, in response to chitin.

Material and methods: Colloidal chitin was added to induce the culture of A. oligospora, and the phenotypic differences before and after induction were observed under inverted microscope. The differential proteins before and after mycelium induction were screened by liquid chromatography-tandem mass spectrometry. The differentially expressed chitinase was expressed in Pichia yeast, and the recombinant enzyme was incubated with Caenorhabditis elegans and its egg suspension to explore its biological activity.

Results: It was found that there was a significant acceleration in the mycelial growth post chitin interaction in A. oligospora. A total of 1,124 differentially expressed proteins (DEPs) were identified between the control group (AO-c) and the experimental group (AO-e), with 183 upregulated and 941 downregulated. Gene Ontology analysis revealed that the DEPs acted in various metabolic processes with catalysis and binding functions. Kyoto Encyclopedia of Genes and Genomes analysis associated these proteins primarily with signalling pathways related to glucose metabolism. Three chitinases were significantly modulated among DEPs. Moreover, enzymatic activity assays demonstrated that one of them effectively degraded C. elegans and its eggs.

Conclusion: These findings suggest that A. oligospora can significantly alter its protein expression profile in response to chitin, thereby facilitating its sugar metabolism and mycelial development. Our study provided new insights into the regulatory mechanisms of nematode predation in A. oligospora.

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来源期刊
Journal of Veterinary Research
Journal of Veterinary Research Veterinary-General Veterinary
CiteScore
0.90
自引率
5.60%
发文量
58
审稿时长
18 weeks
期刊介绍: Journal of Veterinary Research (formerly Bulletin of the Veterinary Institute in Pulawy) is a quarterly that publishes original papers, review articles and short communications on bacteriology, virology, parasitology, immunology, molecular biology, pathology, toxicology, pharmacology, and biochemistry. The main emphasis is, however, on infectious diseases of animals, food safety and public health, and clinical sciences.
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