基于非靶向代谢组学的平菇子实体发育过程中菌丝体差异代谢物分析

Analysis of differential metabolites during fruiting body development of Pleurotus ostreatus based on untargeted metabolomics

为了解平菇生长发育过程中子实体形成的代谢物基础,采用超高效液相色谱-电喷雾串联四级杆质谱(UPLC-ESI-MS/MS)技术结合多变量统计分析方法对发菌完成期(MM)、原基期(MP)及子实体分化期(MF)的平菇菌丝体进行代谢组学分析。结果表明,主成分(PCA)模型分析结果显示3个时期平菇菌丝体中的代谢产物具有明显差异。通过正交偏最小二乘判别分析(OPLS‐DA),以VIP(varible importance in the projection)>1和差异倍数值(fold change)≥2或≤0.5为条件对MM vs MP、MM vs MF和MP vs MF中的差异代谢物进行比较分析,分别获得139个、147个和67个差异代谢物,变化倍数最大的物质包括氨基酸及其衍生物、脂质、生物碱、有机酸等,说明这些差异代谢物对平菇子实体发育具有重要影响。KEGG分析表明,苯丙氨酸代谢、色氨酸代谢、嘌呤代谢等20条代谢通路表现活跃。在子实体发育过程中,脂质、有机酸、核苷酸及其衍生物、氨基酸及其衍生物之间明显相关。以上研究结果为平菇子实体发育机制和标准化栽培提供了理论依据。

In order to understand the metabolite basis of fruiting body formation during the growth and development of Pleurotus ostreatus,ultra performance liquid chromatography-tandem mass spectrometry interfaced with electro-spray ionization(UPLC-ESI-MS/MS)combined with multivariate statistical methods was used to analyze the differences of intracellular metabolites in mycelia of P.ostreatus at the spawning completion stage,primordium stage and fruiting body differentiation stage.The results showed that principal component(PCA)model revealed significant differences(p<0.05)among metabolites in mycelia of P.ostreatus at the three periods.Through orthogonal partial least squares discriminant analysis(OPLS‐DA),139,147 and 67 differential metabolites were screened and identified in MM vs MP,MM vs MF and MP vs MF under the conditions of VIP(varible importance in the projection)>1 and fold change≥2 or≤0.5.Amino acids and derivatives,lipids,alkaloids and organic acids were substances with the maximum fold change values,which indicated that these differential metabolites played important roles in the fruiting body development.KEGG analysis showed that phenylalanine metabolism,tryptophan metabolism,purine metabolism and other metabolic pathways were active.During the growth and development of P.ostreatus,the contents of lipids,organic acids,nucleotides and their derivatives,and amino acids and their derivatives were significantly correlated.The results of this study provided theoretical basis for the mechanism of fruiting body development and standardized cultivation of P.ostreatus.