双孢蘑菇子实体发育不同阶段蛋白质组分析

Developmental proteomics analysis of the button mushroom Agaricus bisporus

为探讨双孢蘑菇子实体发育不同阶段的蛋白质表达变化,对双孢蘑菇As2796菌株子实体原基期、幼菇期、采收期、开伞期的蛋白质组进行了二维液相色谱串联质谱(i TRAQ-MS/MS)分析,共获得不同肽段5 869个,鉴定到1 059个蛋白质,其中1 007个具有相对定量信息。与双孢蘑菇原基期相比,幼菇期、采收期和开伞期分别有差异蛋白质242、200、240个,分别占鉴定蛋白质总数的24.0%,19.9%和23.8%。对这些蛋白质及不同阶段之间的差异蛋白质进行了系列生物信息学分析,对8个上、下调表达具有连续性的差异蛋白质相关基因进行了荧光定量PCR验证,其中3个蛋白质(错配碱基识别蛋白、细胞色素C1及某推定的未知蛋白质)基因的转录与蛋白质的表达具有较为一致的趋势。这些结果为后续双孢蘑菇子实体发育相关基因的功能研究奠定了良好的基础。

In order to reveal the protein expression changes that occur at the different developmental stages of the button mushroom Agaricus bisporus fruiting body, the technique of iTRAQ-MS/MS （i.e. isobaric tags for relative and absolute quantification coupled two-dimensional liquid chromatography tandem mass spectrometry） was used to analyze the fruiting body proteomes of the primordium, button, harvest and opening stages of A. bisporus As2796 strain. As a result, 1 059 proteins, included a total of 5 869 unique peptides, were identified. Among them, 1 007 proteins were detected with quantitative information. As compared with the proteins of primordium stage, the samples of button, harvest and opening stages produced 242, 200, and 240 different proteins, respectively, accounting for 24.0%, 19.9% and 23.8% of the proteins identified, respectively Meanwhile, the differentially expressed proteins between different stages, as well as all of the identified proteins, were analyzed via bioinformatics. The genes of 8 proteins either up- or down-expressed continuously at different developmental stages were analyzed by real-time PCR （qPCR）, and 3 of them, i.e., mismatched base pair and cruciform DNA recognition protein, Cytochrome C2 and a hypothetical protein, were confirmed to have similar expression trends with the corresponding proteins. The data of this study provided a basis for future functional research of the genes involved in fruiting body development in A. bisporus