云南黑松露及其共生松根的物质及通路关联分析

Correlation analyses of substances and pathways of Tuber sinenses and their symbiotic pine roots

摘要:

松露必须与植物共生形成外生菌根，通过与菌根进行物质互利交换才能形成子实体。【目的】云南黑松露(Tuber sinense,SL)多与云南松(Pinus yunnanensis)共生，探明云南松露与其共生松树的物质交流。【方法】应用液相质谱(liquid chromatograph mass spectrometer,LC-MS)技术，对新鲜的云南黑松露及云南松松根(Pinus yunnanensis roots,SG)的代谢物谱进行分析。【结果】在SL及SG中分别检测到1 304个和1 516个物质，其中SL特有物质(Tuber sinense specificity,SLS)399个，SG特有物质(Pinus yunnanensis roots specificity,SGS)611个；二者共有的显著差异物质294个，包括SL上调物质(differential metabolites-up,DEMs-up)93个，SL下调物质201个。网络相关分析表明，相对含量≥2.0的100个(SLS及SGS 40个，DEMs-up 60个)物质有92个存在显著相关性。SLS及SGS中29个关联物质及DEMs-up中19个物质，富集到15条重要通路，其主要涉及次生代谢产物的生物合成、氨基酸的生物合成及代谢、维生素的生物合成及代谢三大类通路。【结论】松露及松根所含有的代谢物间存在一定的网络关联；松露及松根中含有的一些物质，包括环磷酸腺苷(cAMP)、植物甾醇、3-吲哚乙酸等物质对子实体的发育具有重要的调控作用。研究结果为今后分析与子实体发育相关的潜在信号分子及探明云南松露与其共生松树的共生机制提供了理论依据，同时为筛选菌根苗及子实体的促生物质，为规模化人工培养松露等奠定了实践基础。

Abstract:

Truffles must be symbiotic with plants to form ectomycorrhiza (ECM), which facilitates the formation of fruiting bodies by mutually beneficial exchanges of substances. [Objective] To elucidate the flow of substances between Tuber sinense and Pinus yunnanensis in a symbiosis relationship. [Methods] Liquid chromatography-mass spectrometry (LC-MS) was employed to analyze the metabolite profiles of freshly harvested T. sinense (SL) and P. yunnanensis roots (SG). [Results] In SL and SG, 1 304 and 1 516 substances were detected respectively, including 399 SL-specific substances (SLSs) and 611 SG-specific substances (SGSs). There were 294 common differential substances, including 93 up-regulated substances (DEMs-up) and 201 down-regulated substances in SL. The network correlation analysis revealed that 92 of the 100 substances with the relative content ≥2.0 (40 SLSs and SGSs and 60 DEMs-up) displayed significant correlations. Twenty-nine associated substances in SLSs and SGSs and 19 substances in DEMs-up were enriched in 15 important pathways, which mainly involved secondary metabolite biosynthesis, amino acid biosynthesis and metabolism, and vitamin biosynthesis and metabolism. [Conclusion] A network correlation existed between T. sinense and P. yunnanensis pine roots metabolites. Some substances in truffles and pine roots, including adenosine 3',5'-cyclic monophosphate (cAMP), phytosterol, and indole-3-acetic acid, had regulatory effects on the development of fruiting bodies. The findings provided a theoretical basis for further studying the potential signaling molecules associated with fruiting body development and elucidating the symbiotic mechanism between T. sinense and pines. Additionally, this study laid a practical basis for screening the substances promoting the growth of mycorrhizal seedlings and fruiting bodies and for large-scale artificial cultivation of truffles.