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iTRAQ-MS/MS蛋白质组学分析揭示双孢蘑菇采后成熟过程中的差异表达蛋白。

iTRAQ-MS/MS Proteomic Analysis Reveals Differentially Expressed Proteins During Post-harvest Maturation of the White Button Mushroom Agaricus bisporus.

作者信息

Chen Meiyuan, Liao Jianhua, Li Hongrong, Cai Zhixin, Guo Zhongjie, Wach Mark P, Wang Zesheng

机构信息

The Institute of Edible Fungi, Fujian Academy of Agricultural Sciences, No.10, 95 Lane, Qianheng Rd., Fuzhou, 350014, People's Republic of China.

Sylvan Research, West Hills Industrial Park, Bldg 2, Kittanning, PA, 16201, USA.

出版信息

Curr Microbiol. 2017 May;74(5):641-649. doi: 10.1007/s00284-017-1225-y. Epub 2017 Mar 16.

DOI:10.1007/s00284-017-1225-y
PMID:28299440
Abstract

Agaricus bisporus (J. E. Lange) Imbach, well known as the common cultivated mushroom or white button mushroom, is widely cultivated all over the world. We used iTRAQ-MS/MS (i.e., isobaric tags for relative and absolute quantification-coupled two-dimensional liquid chromatography-tandem mass spectrometry) to identify protein expression changes that occur during the post-harvest maturation of Agaricus bisporus fruitbodies at 0, 6, 12, and 48 h after harvest. A total of 5878 unique peptides, representing 1063 proteins, were identified. Quantitative data were obtained from 1012 out of the 1063 proteins. A total of 102, 106, and 160 differentially expressed proteins (>twofolds differences) were identified from samples collected at 6, 12, and 48 h compared to sample from 0 h post harvest, accounting for 10.1, 10.5, and 15.8% of the total proteins identified, respectively. All identified proteins including the differentially expressed proteins among different post-harvest stages were subjected to bioinformatic analysis. Furthermore, seven representative proteins either up or down-regulated at different post-harvest stages were analyzed by real-time PCR. Three out of the seven proteins, the mismatched base pair and cruciform DNA recognition protein, hydrophobin-B and protein transporter, exhibited similar expression patterns as their corresponding proteins. The 260 differentially expressed proteins identified from our study laid a foundation for future studies aiming to understand the post-harvest maturation process of A. bisporus and eventually, might help in the development of breeding program to identify strains with extended shelf life.

摘要

双孢蘑菇(学名:Agaricus bisporus (J. E. Lange) Imbach),即众所周知的普通栽培蘑菇或白蘑菇,在全球广泛种植。我们使用iTRAQ-MS/MS(即相对和绝对定量等压标签结合二维液相色谱-串联质谱法)来鉴定双孢蘑菇子实体在收获后0、6、12和48小时的采后成熟过程中发生的蛋白质表达变化。共鉴定出5878个独特肽段,代表1063种蛋白质。从1063种蛋白质中的1012种获得了定量数据。与收获后0小时的样品相比,在收获后6、12和48小时采集的样品中分别鉴定出102、106和160种差异表达蛋白质(差异>两倍),分别占鉴定出的总蛋白质的10.1%、10.5%和15.8%。对所有鉴定出的蛋白质,包括不同采后阶段的差异表达蛋白质进行了生物信息学分析。此外,对在不同采后阶段上调或下调的7种代表性蛋白质进行了实时PCR分析。7种蛋白质中的3种,即错配碱基对和十字形DNA识别蛋白、疏水蛋白-B和蛋白质转运蛋白,表现出与其相应蛋白质相似的表达模式。我们研究中鉴定出的260种差异表达蛋白质为未来旨在了解双孢蘑菇采后成熟过程的研究奠定了基础,并最终可能有助于开发育种计划以鉴定货架期延长的菌株。

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