基于nano LC-MS/MS技术草酸影响猪苓菌核形成的乙酰化修饰蛋白质组学研究

批准号:
81973425
项目类别:
面上项目
资助金额:
55.0 万元
负责人:
邢咏梅
依托单位:
学科分类:
中药资源
结题年份:
2023
批准年份:
2019
项目状态:
已结题
项目参与者:
邢咏梅
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中文摘要
猪苓是珍贵的药用真菌,菌核入药,在我国用药历史悠久。然而目前野生菌核资源逐渐贫乏。本项目在发现内源性草酸在菌丝和菌核的含量存在显著差异且与氧化应激密切相关及不同浓度外源性草酸可促进或抑制菌核形成后,以nano LC-MS/MS乙酰化修饰蛋白质组学技术获得乙酰化修饰蛋白标记物并阐述草酸/氧化应激影响猪苓菌核发育中能量代谢的分子机理;利用免疫胶体金技术、Western blot及MRM定量检测技术研究在猪苓菌核与菌丝间差异表达且与细胞间粘连及细胞骨架组成密切相关的integrin ß1,actin与tubulin的亚细胞结构定位与分布及表达情况,揭示其在菌丝融合、细胞骨架重塑中的作用。应用非损伤微测技术对草酸影响猪苓菌核形成中Ca2+与H2O2进行定量实时监测,揭示Ca2+与活性氧的变化规律及菌核形成中草酸与Ca2+、H2O2变化的关系。本项目将为草酸影响猪苓菌核发育的分子机制奠定基础。
英文摘要
Polyporus umbellatus is one of the most precious medicinal fungi, which has been used as medicine for more than two thousand years. However, wild sclerotia of P. umbellatus are almost in danger of extinction now..This study is based on our previous study which indicates that endogenous oxalic acid content differs very much in P. umbellatus mycelia and sclerotia and it is closely associated to oxidative stress. Furthermore, different concentrations of exogenous oxalic acid affect sclerotial formation at different manners. The study conducts to investigate the differential genes affected by oxalic acid using nano LC-MS/MS acetylated modification proteomics technology, mainly paying attention to oxalic acid and oxidative stress affecting energy metabolism regulation during P. umbellatus sclerotial formation and acquiring protein markers after acetylated modification. In the previous study, integrin ß1, actin and tubulin were found to be significantly and differentially expressed in mycelia and sclerotia during P. umbellatus sclerotial formation, which were closely associated with adhesion between cells and being main components of cytoskeleton. Based on their subcellular localization, distribution and expression characteristics using Immunogold technique, western blot and quantitative MRM detection assay, it will facilitate to discover what roles integrin ß1, actin and tubulin will play during mycelia fusion and cytoskeleton remodeling affected by oxalic acid. Furthermore, Ca2+ and H2O2 are quantitative real-time dynamic monitored using Non-invasive Micro-test Technology during different stages of sclerotial formation affected by oxalic acid, which will help to elucidate the changing rules of Ca2+ and ROS and the relationship among oxalic acid and changes of Ca2+ and ROS. This study will provide us information for molecular mechanism induced by oxalic acid in P. umbellatus sclerotial formation.
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DOI:10.16438/j.0513-4870.2019-0900
发表时间:2020
期刊:药学学报
影响因子:--
作者:刘蒙蒙;邢咏梅;郭顺星
通讯作者:郭顺星
Armillaria mellea Symbiosis Drives Metabolomic and Transcriptomic Changes in Polyporus umbellatus Sclerotia.
蜜环菌共生驱动猪苓菌核的代谢组学和转录组学变化
DOI:10.3389/fmicb.2021.792530
发表时间:2021
期刊:Frontiers in microbiology
影响因子:5.2
作者:Xing YM;Li B;Liu L;Li Y;Yin SX;Yin SC;Chen J;Guo SX
通讯作者:Guo SX
Use of transcriptomic profiling to identify candidate genes involved in Polyporus umbellatus sclerotial formation affected by oxalic acid.
使用转录组分析来鉴定参与受草酸影响的猪苓菌核形成的候选基因
DOI:10.1038/s41598-021-96740-7
发表时间:2021-08-30
期刊:Scientific reports
影响因子:4.6
作者:Xing YM;Li B;Zeng X;Zhou LS;Lee TS;Lee MW;Chen XM;Guo SX
通讯作者:Guo SX
DOI:10.13344/j.microbiol.china.210480
发表时间:2022
期刊:微生物学通报
影响因子:--
作者:李兵;刘柳;单婷婷;邢咏梅;郭顺星
通讯作者:郭顺星
药用真菌猪苓菌丝形成菌核差异基因及功能的研究
- 批准号:31201666
- 项目类别:青年科学基金项目
- 资助金额:20.0万元
- 批准年份:2012
- 负责人:邢咏梅
- 依托单位:
国内基金
海外基金
