Genome-Wide Analysis of Terpene Synthase Genes in Crocus sativus Reveals Their Regulatory Roles in Terpenoid Biosynthesis and Abiotic Stress Tolerance

Terpene synthases (TPS) facilitate terpenoid production, influencing the flavor, color, and medicinal properties of <i>Crocus sativus</i> (saffron), a triploid geophyte of significant commercial importance. Despite its importance, the <i>CsTPS</i> gene family remains poorly characterized, limiting genetic enhancements in saffron's agronomic features. This research performed a comprehensive genome-wide analysis of <i>CsTPS</i> genes using genomic, transcriptomic, and in silico approaches. BLASTP and PfamScan discovered thirty <i>CsTPS</i> genes, demonstrating conserved TPS domains, varied exon-intron architectures, and chromosomal clustering indicative of tandem duplications. Phylogenetic research categorized these genes into five subfamilies (TPS-a to TPS-e), with the prevalence of TPS-a suggesting a role in sesquiterpene biosynthesis. RNA-seq data (PRJNA976833, PRJNA400472) revealed tissue-specific expression, with <i>CsTPS1</i> and <i>CsTPS5</i> expressed in reproductive tissues and <i>CsTPS2</i> in vegetative tissues. Stress-responsive genes (<i>CsTPS1</i>, <i>CsTPS4</i>) exhibited upregulation in response to cold and pathogen stress, with cis-regulatory elements (e.g., ARE, ABRE) indicating hormone control. The in-silico validation of <i>CsTPS1</i>, chosen for its elevated GMQE score (0.89), included primer design, ePCR, and vector optimization for expression in <i>Arabidopsis thaliana</i>. This study elucidates the contribution of the <i>CsTPS</i> family to saffron terpenoid diversity, providing a foundation for enhancing flavor, yield, and stress tolerance through genetic engineering.