Table_6_Integrated physiological, metabolomic, and proteome analysis of Alpinia officinarum Hance essential oil inhibits the growth of Fusarium oxysporum of Panax notoginseng.XLSX

Fusarium oxysporum is the main pathogen of Panax notoginseng root rot, and chemical fungicides remain the primary measures to control the disease. Plant essential oil (EO) is a volatile plant secondary metabolic product that does not produce any residue to replace chemical pesticide. To comprehensively understand the antifungal mechanism of Alpinia officinarum Hance EO, the physiological indicators, proteome and metabolome were analyzed using F. oxysporum spores and hyphae treated with different EO concentrations. The cell membrane was damaged after both low and high concentrations of EO treatment, along with leakage of the cell contents. To resist the destruction of membrane structure, fungi can increase the function of steroid biosynthesis and expression of these catalytic enzymes, including squalene monooxygenase (SQLE), sterol 14alpha-demethylase (CYP51, CYP61A), delta14-sterol reductase (TM7SF2, ERG4), methylsterol monooxygenase (MESO1), and sterol 24-C-methyltransferase (SMT1). Furthermore, the tricarboxylic acid cycle (TCA) was influenced by inhibiting the expression of glutamate synthase (GLT1), 4-aminobutyrate aminotransferase (ABAT), and succinate-semialdehyde dehydrogenase (gabD); increasing malate and gamma-aminobutyric acid (GABA); and decreasing citrate content. The spore germination rate and mycelia growth were decreased because the expression of cohesin complex subunit SA-1/2 (IRR1) and cohesion complex subunit (YCS4, BRN1, YCG1) were inhibited. Particularly, under high EO concentrations, cyclin-dependent kinase (CDC28) and DNA replication licensing factor (MCM) were further inhibited to disrupt the cell cycle and meiosis, thus affecting cell division. The results of this study will enrich the understanding of the antifungal mechanism of EOs and provide an important basis to develop new plant-derived fungicides.

尖孢镰刀菌（Fusarium oxysporum）是三七根腐病的主要病原体，化学杀菌剂仍然是控制该病的主要措施。植物精油（EO）作为一种挥发性植物次级代谢产物，不产生任何残留，可替代化学农药。为全面理解高良姜精油（Alpinia officinarum Hance EO）的抑真菌机制，本研究采用不同浓度的精油处理尖孢镰刀菌孢子及菌丝，分析了生理指标、蛋白质组学和代谢组学。结果显示，无论是低浓度还是高浓度的精油处理，均导致细胞膜受损，细胞内容物泄漏。为了抵抗细胞结构的破坏，真菌能够增强甾体生物合成功能以及以下催化酶的表达：单氧合酶（squalene monooxygenase, SQLE）、甾醇14α-去甲基化酶（CYP51, CYP61A）、Δ14-甾醇还原酶（TM7SF2, ERG4）、甲基甾醇单氧合酶（MESO1）和甾醇24-C-甲基转移酶（SMT1）。此外，通过抑制谷氨酸合成酶（GLT1）、γ-氨基丁酸转氨酶（ABAT）和琥珀酸半醛脱氢酶（gabD）的表达，增加苹果酸和γ-氨基丁酸（GABA）的含量，以及降低柠檬酸含量，影响了三羧酸循环（TCA）。由于抑制了粘合蛋白复合体亚基SA-1/2（IRR1）和粘合蛋白复合体亚基（YCS4, BRN1, YCG1）的表达，孢子发芽率和菌丝生长均受到抑制。特别是在高浓度精油作用下，周期依赖性激酶（CDC28）和DNA复制许可因子（MCM）进一步受到抑制，从而干扰细胞周期和减数分裂，影响细胞分裂。本研究结果将丰富对植物精油抑真菌机制的理解，并为开发新型植物源杀菌剂提供重要的理论依据。