Table_5_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.

尖孢镰刀菌是三七根腐病的主要病原菌，化学杀菌剂仍为控制该病的主要手段。植物精油（EO）作为一种挥发性次生代谢产物，不产生任何残留，可替代化学农药。为全面理解高良姜（Alpinia officinarum Hance）精油抗真菌作用机制，本研究采用尖孢镰刀菌的孢子及菌丝，分别用不同浓度的精油处理，对其生理指标、蛋白质组学和代谢组学进行了分析。结果显示，低浓度与高浓度的精油处理均导致细胞膜损伤及细胞内容物泄漏。为抵抗细胞结构的破坏，真菌可通过增加甾体生物合成功能及催化酶（包括鲨烯单加氧酶（SQLE）、甾醇14α-脱甲基酶（CYP51、CYP61A）、delta14-甾醇还原酶（TM7SF2、ERG4）、甲基甾醇单加氧酶（MESO1）和甾醇24-C-甲基转移酶（SMT1）的表达）来维持膜结构的稳定。此外，通过抑制谷氨酸合成酶（GLT1）、4-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，降低柠檬酸含量，影响了三羧酸循环（TCA）。此外，抑制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，降低了孢子发芽率和菌丝生长。在高浓度精油的作用下，细胞周期和减数分裂进一步受到抑制，这归因于细胞周期依赖性激酶（CDC28）和DNA复制许可因子（MCM）的活性被抑制。本研究结果将丰富对植物精油抗真菌作用机制的理解，并为开发新型植物源性杀菌剂提供了重要依据。