title: Combined metabolome and transcriptome profiling provides new insights into diterpene biosynthesis in S. pomifera glandular trichomes.
abstract: BACKGROUND
  Salvia diterpenes have been found to have health promoting properties. Among them, carnosic acid and carnosol, tanshinones and sclareol are well known for their cardiovascular, antitumor, antiinflammatory and antioxidant activities. However, many of these compounds are not available at a constant supply and developing biotechnological methods for their production could provide a sustainable alternative. The transcriptome of S.pomifera glandular trichomes was analysed aiming to identify genes that could be used in the engineering of synthetic microbial systems.
  
  
  RESULTS
  In the present study, a thorough metabolite analysis of S. pomifera leaves led to the isolation and structure elucidation of carnosic acid-family metabolites including one new natural product. These labdane diterpenes seem to be synthesized through miltiradiene and ferruginol. Transcriptomic analysis of the glandular trichomes from the S. pomifera leaves revealed two genes likely involved in miltiradiene synthesis. Their products were identified and the corresponding enzymes were characterized as copalyl diphosphate synthase (SpCDS) and miltiradiene synthase (SpMilS). In addition, several CYP-encoding transcripts were identified providing a valuable resource for the identification of the biosynthetic mechanism responsible for the production of carnosic acid-family metabolites in S. pomifera.
  
  
  CONCLUSIONS
  Our work has uncovered the key enzymes involved in miltiradiene biosynthesis in S. pomifera leaf glandular trichomes. The transcriptomic dataset obtained provides a valuable tool for the identification of the CYPs involved in the synthesis of carnosic acid-family metabolites.
authors.affiliation: Institute of Applied Biosciences/CERTH, P.O. Box 60361, Thermi, 57001, , Thessaloniki, Greece. ftrikka@gmail.com.
authors.forename: Fotini A
authors.initials: FA
authors.lastname: Trikka
authors.affiliation: Institute of Applied Biosciences/CERTH, P.O. Box 60361, Thermi, 57001, , Thessaloniki, Greece. nikolchem@hotmail.com.
authors.forename: Alexandros
authors.initials: A
authors.lastname: Nikolaidis
authors.affiliation: Department of Biochemistry, School of Medicine, University of Crete, P.O. Box 2208, Heraklion, 71003, Greece. igneacodruta@yahoo.com.
authors.forename: Codruta
authors.initials: C
authors.lastname: Ignea
authors.affiliation: Institute of Applied Biosciences/CERTH, P.O. Box 60361, Thermi, 57001, , Thessaloniki, Greece. afroditi.tsaballa@gmail.com.
authors.forename: Aphrodite
authors.initials: A
authors.lastname: Tsaballa
authors.affiliation: Department of Pharmacognosy and Chemistry of Natural Products, Faculty of Pharmacy, University of Athens, Panepistimiopolis Zografou, Athens, 15771, Greece. ltziveleka@pharm.uoa.gr.
authors.forename: Leto-Aikaterini
authors.initials: LA
authors.lastname: Tziveleka
authors.affiliation: Department of Pharmacognosy and Chemistry of Natural Products, Faculty of Pharmacy, University of Athens, Panepistimiopolis Zografou, Athens, 15771, Greece. eioannou@pharm.uoa.gr.
authors.forename: Efstathia
authors.initials: E
authors.lastname: Ioannou
authors.affiliation: Department of Pharmacognosy and Chemistry of Natural Products, Faculty of Pharmacy, University of Athens, Panepistimiopolis Zografou, Athens, 15771, Greece. roussis@pharm.uoa.gr.
authors.forename: Vassilios
authors.initials: V
authors.lastname: Roussis
authors.affiliation: Institute of Applied Biosciences/CERTH, P.O. Box 60361, Thermi, 57001, , Thessaloniki, Greece. elenstea@gmail.com.
authors.forename: Eleni A
authors.initials: EA
authors.lastname: Stea
authors.affiliation: Group of Biotechnology of Pharmaceutical Plants, Laboratory of Pharmacognosy, Department of Pharmaceutical Sciences, Aristotle University of Thessaloniki, 541 24, Thessaloniki, Greece. dragana.bozic@ibiss.bg.ac.rs.|Institute for Biological Research "Siniša Stanković", University of Belgrade, Belgrade, Serbia. dragana.bozic@ibiss.bg.ac.rs.
authors.forename: Dragana
authors.initials: D
authors.lastname: Božić
authors.affiliation: Institute of Applied Biosciences/CERTH, P.O. Box 60361, Thermi, 57001, , Thessaloniki, Greece. argiriou@certh.gr.
authors.forename: Anagnostis
authors.initials: A
authors.lastname: Argiriou
authors.affiliation: Group of Biotechnology of Pharmaceutical Plants, Laboratory of Pharmacognosy, Department of Pharmaceutical Sciences, Aristotle University of Thessaloniki, 541 24, Thessaloniki, Greece. kanellis@pharm.auth.gr.
authors.forename: Angelos K
authors.initials: AK
authors.lastname: Kanellis
authors.affiliation: Department of Biochemistry, School of Medicine, University of Crete, P.O. Box 2208, Heraklion, 71003, Greece. s.kampranis@med.uoc.gr.
authors.forename: Sotirios C
authors.initials: SC
authors.lastname: Kampranis
authors.affiliation: Institute of Applied Biosciences/CERTH, P.O. Box 60361, Thermi, 57001, , Thessaloniki, Greece. antoniosmakris@yahoo.gr.
authors.forename: Antonios M
authors.initials: AM
authors.lastname: Makris
mesh_terms: D003577:Cytochrome P-450 Enzyme System / Q000145:classification / Q000235:genetics; D004224:Diterpenes / Q000378:metabolism; D055442:Metabolome / Q000235:genetics*; D058977:Molecular Sequence Annotation; D015394:Molecular Structure; D018515:Plant Leaves / Q000378:metabolism; D027544:Salvia / Q000235:genetics* / Q000378:metabolism; D013729:Terpenes / Q000737:chemistry / Q000378:metabolism*; D059467:Transcriptome / Q000235:genetics*; D064201:Trichomes / Q000235:genetics*
chemical_list: D004224:Diterpenes; D013729:Terpenes; C573600:miltiradiene; D003577:Cytochrome P-450 Enzyme System