广叶绣球菌不同生长阶段栽培基质中代谢物差异分析

Metabolite differences in cultivation substrates of Sparassis latifolia at different developmental stages

摘要:

【背景】栽培基质的利用是广叶绣球菌(Sparassis latifolia)栽培中重要的生理过程，但栽培过程中基质代谢物的变化尚不清楚。【目的】通过不同生长阶段栽培基质中差异代谢物分析挖掘关键代谢物，为广叶绣球菌基质利用机理研究提供理论参考。【方法】利用UHPLC-MS/MS技术分析广叶绣球菌菌丝(Myc)、原基(Pri)和子实体(FB)生长阶段栽培基质中代谢产物的变化，通过不同数据库进行代谢物注释并进行KEGG通路富集分析。利用LC-MS/MS技术检测不同发育阶段绣球菌中植物类激素含量。【结果】三个不同栽培阶段基质中共鉴定出代谢产物1 360个。不同比较组(Pri vs. Myc、FB vs. Myc和FB vs. Pri)间共有的差异代谢产物179个，含量最高的50个代谢物主要包括氨基酸、脂质、吡喃酸、吡喃酮和植物类激素等物质。其中氨基酸含量在Myc、Pri和FB阶段基质中逐渐降低，而吡喃酸和吡喃酮类化合物含量逐渐升高。植物类激素中的赤霉素在Pri和FB阶段基质中含量较高，茉莉酸在Myc阶段基质中含量较高。对不同发育阶段绣球菌植物类激素进行检测，发现赤霉素GA7仅在原基中检测到，12-氧代植物二烯酸在子实体中含量显著升高。不同比较组所有的差异代谢物KEGG分析显示嘌呤代谢、鞘脂信号通路和甘油磷脂代谢等通路显著富集。【结论】氨基酸是Myc阶段基质中重要的代谢物，吡喃酸、吡喃酮和植物类激素可能与Pri和FB阶段基质利用过程中的生理生化调控有关。

Abstract:

[Background] Substrate utilization is an important physiological process in the cultivation of Sparassis latifolia, while little is known about the metabolite differences in the substrates at different developmental stages.[Objective] To identify the differential metabolites in the cultivation substrates of S. latifolia at different developmental stages, explore key metabolites, and provide a theoretical basis for the research on the mechanism of substrate utilization. [Methods] Ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) was employed to determine the changes of metabolites in the cultivation substrates of mycelium (Myc), primordium (Pri), and fruiting body (FB). Functional annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment were conducted for the metabolites. The content of phytohormones in S. latifolia was determined by liquid chromatography-tandem mass spectrometry (LC-MS/MS). [Results] A total of 1 360 metabolites were identified from the substrates of S. latifolia at three different stages. There were 179 common differential metabolites among three different comparison groups (Pri vs. Myc, FB vs. Myc, and FB vs. Pri), and the top 50 most abundant metabolites were mainly amino acids, lipids, pyranic acids, pyranones, and phytohormones. The content of amino acids gradually decreased while that of pyranic acids and pyranones gradually increased with the development of S. latifolia. The content of gibberellins (GAs) was higher in the substrates of Pri and FB, and that of jasmonic acid was higher in the substrate of Myc. GA7 was detected only in Pri, and the content of 12-oxo-phytodienoic acid was higher in FB. The differential metabolites were mainly enriched in purine metabolism, sphingolipid, glycerophospholipid metabolism pathways. [Conclusion] Amino acids are key metabolites in the substrate of Myc. pyranoic acids, pyranones, and phytohormones may be involved in the physiological and biochemical regulation of substrate utilization in the Pri and FB stages.