| Summary: | <i>Zephyranthes candida</i> is a frequently cultivated ornamental plant containing several secondary metabolites, including alkaloids, flavonoids, and volatile organic compounds (VOCs). However, extensive research has been conducted only on non-VOCs found in the plant, whereas the production of VOCs and the molecular mechanisms underlying the biosynthesis of terpenes remain poorly understood. In this study, 17 volatile compounds were identified from <i>Z. candida</i> flowers using gas chromatography–mass spectrometry (GC-MS), with 16 of them being terpenoids. Transcriptome sequencing resulted in the identification of 17 terpene synthase (<i>TPS</i>) genes; two <i>TPS</i> genes, ZcTPS01 and ZcTPS02, had high expression levels. Biochemical characterization of two enzymes encoded by both genes revealed that <i>ZcTPS02</i> can catalyze geranyl diphosphate (GPP) into diverse products, among which is <i>β</i>-ocimene, which is the second most abundant compound found in <i>Z. candida</i> flowers. These results suggest that <i>ZcTPS02</i> plays a vital role in <i>β</i>-ocimene biosynthesis, providing valuable insights into terpene biosynthesis pathways in <i>Z. candida</i>. Furthermore, the expression of ZcTPS02 was upregulated after 2 h of methyl jasmonate (MeJA) treatment and downregulated after 4 h of the same treatment.
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