title: Elucidation of terpenoid metabolism in Scoparia dulcis by RNA-seq analysis.
abstract: Scoparia dulcis biosynthesize bioactive diterpenes, such as scopadulcic acid B (SDB), which are known for their unique molecular skeleton. Although the biosynthesis of bioactive diterpenes is catalyzed by a sequence of class II and class I diterpene synthases (diTPSs), the mechanisms underlying this process are yet to be fully identified. To elucidate these biosynthetic machinery, we performed a high-throughput RNA-seq analysis, and de novo assembly of clean reads revealed 46,332 unique transcripts and 40,503 two unigenes. We found diTPSs genes including a putative syn-copalyl diphosphate synthase (SdCPS2) and two kaurene synthase-like (SdKSLs) genes. Besides them, total 79 full-length of cytochrome P450 (CYP450) genes were also discovered. The expression analyses showed selected CYP450s associated with their expression pattern of SdCPS2 and SdKSL1, suggesting that CYP450 candidates involved diterpene modification. SdCPS2 represents the first predicted gene to produce syn-copalyl diphosphate in dicots. In addition, SdKSL1 potentially contributes to the SDB biosynthetic pathway. Therefore, these identified genes associated with diterpene biosynthesis lead to the development of genetic engineering focus on diterpene metabolism in S. dulcis.
authors.affiliation: Graduate School of Medicine and Pharmaceutical Sciences for Research, University of Toyama, 2630 Sugitani, Toyama, Toyama 930-0194, Japan.
authors.forename: Yoshimi
authors.initials: Y
authors.lastname: Yamamura
authors.affiliation: Graduate School of Medicine and Pharmaceutical Sciences for Research, University of Toyama, 2630 Sugitani, Toyama, Toyama 930-0194, Japan.
authors.forename: Fumiya
authors.initials: F
authors.lastname: Kurosaki
authors.affiliation: Graduate School of Medicine and Pharmaceutical Sciences for Research, University of Toyama, 2630 Sugitani, Toyama, Toyama 930-0194, Japan.
authors.forename: Jung-Bum
authors.initials: JB
authors.lastname: Lee
mesh_terms: D053898:Biosynthetic Pathways; D059014:High-Throughput Nucleotide Sequencing; D036721:Scoparia / Q000235:genetics* / Q000378:metabolism*; D017423:Sequence Analysis, RNA; D013729:Terpenes / Q000378:metabolism*
chemical_list: D013729:Terpenes