橙黄乳菇-马尾松菌根广靶代谢组学比较与分析

Comparison and analysis of widely targeted metabolomics of mycorrhizal of Lactarius akahatsu and Pinus massoniana

【目的】分析比较橙黄乳菇-马尾松菌根合成前后根系代谢物差异,为探究调控橙黄乳菇-马尾松共生的潜在根系代谢物提供参考。【方法】在纯培养条件下进行橙黄乳菇-马尾松菌根合成,基于液质联用技术测定马尾松及橙黄乳菇-马尾松菌根根系代谢物成分,利用KEGG和HMDB数据库进行代谢物的定量与注释,采用主成分分析、正交偏最小二乘-判别分析等多元统计方法进行代谢组数据的比较,并筛选差异代谢物。【结果】共检测出代谢物627种,其中差异代谢物95种,上调39种,下调56种,主要集中在黄酮类化合物、萜类化合物、氨基酸及其衍生物、酚类化合物、生物碱类化合物、有机酸及其衍生物、核苷酸及其衍生物、香豆素和木脂素、植物激素等。KEGG代谢通路分析显示次生代谢产物的生物合成、苯丙烷类化合物的生物合成、类黄酮生物合成、神经活性配体-受体相互作用、铁载体基非核糖体肽的生物合成、苯甲酸盐降解途径、植物激素的生物合成、莽草酸途径生物碱的生物合成和氨基苯甲酸降解等9条差异代谢途径在橙黄乳菇-马尾松菌根形成后差异代谢物显著富集,可能对橙黄乳菇-马尾松菌根形成具有重要调节作用。【结论】利用广泛靶向代谢组学技术可在整体水平上揭示橙黄乳菇-马尾松菌根形成前后的代谢组学差异,研究结果可为明晰橙黄乳菇-马尾松共生机制及橙黄乳菇-马尾松菌根合成提供科学依据。

【Objective】This study aims to analyze and compare the differences in root metabolites before and after mycorrhizal synthesis between Lactarius akahatsu and Pinus massoniana,in order to provide reference for exploring the potential substances regulating the symbiosis between L.akahatsu and P.massoniana.【Method】The mycorrhizal of L.akahatsu and P.massoniana were synthesized on pure media.Liquid chromatography-mass spectrometry were used to determine the metabolic components of root system of P.massoniana and mycorrhizal of symbiotic system between L.akahatsu and P.massoniana.The metabolites were quantified and annotated using KEGG and HMDB databases.Principal component analysis,orthogonal partial least-squares discriminant analysis and other multivariate statistical methods were used to compare the metabolome data and screen the different metabolic components.【Result】627 metabolites were detected from mycorrhizal between L.akahatsu and P.massoniana,of them 95 ones were differential metabolites,39 ones were up-regulated and 56 were down-regulated.The main metabolites were flavonoids,terpenoids,amino acid and derivatives,phenols,alkaloids,organic acids and derivatives,nucleotide and its derivates,phenylpropanoids and lignan,phytohormone,etc.KEEG metabolic pathway analyses showed that different metabolites were significantly enriched in 9 metabolic pathways.They were biosynthesis of secondary metabolites,biosynthesis of phenylpropanoids,flavonoid biosynthesis,neuroactive ligandreceptor interaction,biosynthesis of siderophore group nonribosomal peptides,benzoate degradation,biosynthesis of plant hormones,biosynthesis of alkaloids derived from shikimate pathway and aminobenzoate degradation,which might be an important regulatory role in the formation of mycorrhizal synthesis between L. akahatsu and P. massoniana.【Conclusion】Using UPLC-MS/MS metabolomics technique, the metabolomics differences of mycorrhizal synthesis between L. akahatsu and P. massoniana can be revealed on an overall level. The results can provide a scientific basis for clarifying the symbiotic mechanism and the mycorrhizal synthesis of L. akahatsu and P. massoniana.