Sequencing and variation of terpene synthase gene (TPS2) as the major gene in biosynthesis of thymol in different Thymus species

Thyme (Thymus spp.) is a valuable genus of Lamiaceae family with different pharmaceutical and food properties. Thymol has also been considered as the major essential oil compound in most of the studied Thymus species. In this research, the gene encoding γ-terpinene synthase (Ttps2) was sequenced in T. vulgaris and in eight Iranian thymes including T. carmanicus, T. daenensis, T. fedtschenkoi, T. kotschyanus, T. migricus, T. pubescens, T. serpyllum, and T. trautvetteri. Genetic relationships based on terpene synthase genes were also determined among the studied species. Rapid Amplification of cDNA Ends (RACE) PCR was done to complete the sequence of all species. The cDNA of the studied species possessed an open reading frame ranging from 1788 to 1794 bp that encode for a protein of 596–598 amino acids, presenting all the conserved motifs characteristics of monoterpene synthases. The taxonomic status of Thymus species was determined based on eight reported sections. The species were classified in three major groups. The first and second group comprised species of Micantes and Mastichina sections. The third cluster included the species belonging to Serpyllum and Pseudothymbra sections. Overall, phylogenetic analysis according to whole sequence of Ttps2 gene can help providing insights in respect to its evolutionary process. Finally, clustering based on the amount of main essential oils components (thymol and carvacrol) was compared with that based on Ttps2 gene classification in the studied Thymus species, showing that clustering is not always in accordance.