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药用真菌猪苓菌丝形成菌核分子机制一直是人们关注的热点问题。本研究首次对猪苓进行了系统的蛋白质组学研究。基于液质联用技术并在数据依赖性采集模式下,成功鉴定了猪苓菌丝形成菌核的初始期、菌核生长期和成熟期的蛋白质组,共鉴定蛋白1 391个(global FDR 1%)。采用先进的SWATH MSALL的非标记定量方式对初始期菌丝和菌核进行了差异蛋白质组学研究,从中定量蛋白1 234个,占鉴定蛋白数量的88.7%。猪苓菌丝形成菌核过程中有378个蛋白差异表达。差异蛋白GO注释结果表明,在猪苓菌丝形成菌核过程中,差异蛋白参与的蛋白功能和生物学过程多样,以催化和结合为主,还参与感知环境刺激、信号转导、电子转移、过氧化活性、菌丝生长及胞壁合成等功能。进一步推测分析表明,猪苓菌丝形成菌核可能是由于缺氧或温差等多种环境胁迫诱导所致,生成活性氧并形成氧化应激状态。信号通过丝裂原活化蛋白激酶通路(MAPK通路)途径向下游传递,可能调控了胞壁蛋白糖基化修饰,进而促进猪苓菌丝极性生长和菌丝形态学变化而形成菌核。KEGG代谢通路分析表明,差异蛋白还主要参与了糖、脂质和氨基酸等初级产物以及甾醇、多聚乙酰等次级代谢物的合成与代谢。因此,对差异蛋白质组的深入解析和研究有利于揭示猪苓菌丝形成菌核的蛋白分子机制,具有较强的理论和现实意义。
Medical fungi Polyporus umbellatus sclerotium molecular mechanism has been a hot issue of concern. This study for the first time on the Polyporus umbellatus system proteomics research. Based on liquid chromatography-mass spectrometry (LC-MS / MS) and data-dependent data collection, the proteomes of the initial, sclerotial and mature stages of the mycelia of M. hyphae were successfully identified and identified a total of 1 391 proteins (global FDR 1 %). A differential proteomic study was carried out on the initial mycelia and sclerotia using the SWATH MSALL non-labeling quantitative method, of which 1 234 were quantified, accounting for 88.7% of the total number of identified proteins. 371 proteins were differentially expressed during the process of mycelium formation in H.pylori. The differential protein GO annotation results showed that in the process of mycelium formation, different protein functions and biological processes are involved in the process of hyphae formation, mainly in catalysis and binding, and are also involved in the perception of environmental stimuli, signal transduction, electron transfer, Peroxide activity, mycelium growth and cell wall synthesis and other functions. Further speculation analysis showed that the formation of mycelium may be due to hypoxia or temperature induced by a variety of environmental stresses, the formation of reactive oxygen species and the formation of oxidative stress. The signal is transmitted downstream through mitogen-activated protein kinase pathway (MAPK pathway), which may regulate the glycosylation of the cell wall protein and further promote the polar growth of mycelia and morphological changes of mycelia to form sclerotia. KEGG metabolic pathway analysis showed that the differential proteins are also mainly involved in the synthesis and metabolism of primary products such as sugars, lipids and amino acids as well as secondary metabolites such as sterols and polyacetals. Therefore, the in-depth analysis and study of differential proteome will be helpful to reveal the protein molecular mechanism of mycelium formation in M. hypoglaucum, which has strong theoretical and practical significance.