| Summary: | Infections caused by <i>Candida albicans</i> (<i>C. albicans</i>) and increasing resistance to commonly used drugs lead to a variety of mucosal diseases and systemic infectious diseases. We previously confirmed that the essential oil of <i>Clausena lansium</i> (Lour.) Skeels seeds (CSEO) had antifungal activity against <i>C. albicans</i>, but the detailed mechanism between the chemical components and antifungal activity is unclear. In this study, a quantitative analysis of five volatile components of CSEO, including sabinene, <i>α</i>-phellandrene, <i>β</i>-phellandrene, 4-terpineol, and <i>β</i>-caryophyllene, was carried out using the gas chromatography–mass spectrometry (GC–MS) method. Both the broth dilution and kinetic growth methods proved that the antifungal activity of CSEO against fluconazole-resistant <i>C. albicans</i> was better than that of its main components (sabinene and 4-terpineol). To further investigate the inhibitory mechanism, the transcriptional responses of <i>C. albicans</i> to CSEO, sabinene, and 4-terpineol treatment were determined based on RNA-seq. The Venn diagram and clustering analysis pattern of differential expression genes showed the mechanism of CSEO and 4-terpineol’s anti-<i>C. albicans</i> activity might be similar from the perspective of the genes. Functional enrichment analysis suggested that CSEO regulated adherence-, hyphae-, and biofilm-formation-related genes, which may be CSEO’s active mechanism of inhibiting the growth of fluconazole-resistant <i>C. albicans</i>. Overall, we preliminarily revealed the molecular mechanism between the chemical components and the antifungal activity of CSEO against <i>C. albicans.</i> This study provides new insights to overcome the azole resistance of <i>C. albicans</i> and promote the development and application of <i>C. lansium</i> (Lour.) Skeels seeds.
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