柞蚕蛹培养蛹虫草不同时间后的代谢组分析

为了解柞蚕蛹培养蛹虫草（简称柞蚕蛹虫草）不同时间后的代谢物变化规律,利用广泛靶向代谢组学技术,比较不同培养时期的柞蚕蛹虫草代谢产物成分,找出差异代谢物并进行代谢通路分析。在柞蚕蛹虫草的5个生长时期共检测到10类421种化合物,主要包括：氨基酸及其衍生物、核苷酸及其衍生物、萜类、酚酸、有机酸、糖及醇类、甾体、脂类、生物碱、醌类等多种化合物。主成分分析（PCA）结果显示,不同生长时期柞蚕蛹虫草的代谢组表现出不同的代谢特征,5个生长时期样本在得分图中可分为3个不同区域,对照（S1）与子座形成初期（S3）归为一区,菌核期（S2）独自归为一区,子座形成期（S4）和子囊壳形成期（S5）归为一区。S2-S5筛选到的特有差异代谢物有36种,包括虫草素、甘露醇等主要活性物质,其中香豆酰腐胺、五羟色胺、γ-生育三烯酚等物质首次被检测到。对不同生长阶段变化排在前20的差异显著代谢成分进行分析,结果表明上下调幅度较大的物质多为有机酸、酚酸、生物碱、核苷酸及其衍生物等次生代谢产物。5个生长时期筛选出的所有差异代谢物共富集在154条代谢通路上,差异代谢物数量富集最多的前5条通路,分别为代谢途径、次生代谢物的生物合成、ABC转运蛋白、嘌呤代谢和氨酰生物合成。整个生长周期中,具有显著影响的通路是嘌呤代谢与丙氨酸、天门冬氨酸和谷氨酸代谢途径。本研究结果可为柞蚕蛹虫草的深入研究与应用提供参考。

Metabolomic analyses of tussah pupa-cultivated Cordyceps militaris at different growth stages

In order to illustrate the change of metabolites of tussah pupa-cultivated Cordyceps militaris at different growth stages, widely-targeted metabolomics technology was applied to compare the metabolite products and the metabolic pathways of different metabolites were analyzed. In total, 421 compounds of 10 classes were detected from five growth stages of C. militaris, including amino acids and their derivatives, terpenoids, phenolic acids, organic acids, steroids, lipids, alkaloids, quinones, nucleotides and their derivatives and so on. Principal component analysis (PCA) showed that the metabolome of cordyceps exhibited different metabolic characteristics at different growth stages. The samples of five growth periods were classified into three different regions in the score map. Control (S1) and initial stroma stage (S3) were classified into the same category, selerotized stage (S2) was classified into one category alone, and selerotium germination stage (S4) and perithecium formation stage (S5) were classified into other category. And 36 specific metabolites were found from S1-S5, including main active substances such as cordycepin and cordycepic acid. Vanillin, 5-hydroxytryptamine and -tocopherol were firstly detected in C. militaris. The top 20 significantly different components at different growth stages belong to secondary metabolites, such as organic acids, phenolic acids, alkaloids, nucleotides and their derivatives, showing significant up-down regulation. All the differentially metabolites screened from five growth stages were enriched in 154 metabolic pathways, of which top five were metabolic pathways, biosynthesis of secondary metabolites, ABC transporters, purine metabolism, and aminoacyl biosynthesis. The pathways with most significant influence were purine, alanine, aspartic acid and glutamate metabolism pathways. This study provides primary reference for the further study and application of C. militaris cultivated on tussah silkworm.