Planta

, Volume 238, Issue 1, pp 191–204 | Cite as

Genomic characterization, molecular cloning and expression analysis of two terpene synthases from Thymus caespititius (Lamiaceae)

  • A. Sofia Lima
  • Jette Schimmel
  • Brigitte Lukas
  • Johannes Novak
  • José G. Barroso
  • A. Cristina Figueiredo
  • Luis G. Pedro
  • Jörg Degenhardt
  • Helena Trindade
Original Article

Abstract

The identification, isolation and functional characterization of two genes encoding two monoterpene synthases—γ-terpinene synthase (Tctps2) and α-terpineol synthase (Tctps5)—from three chemically distinct Thymus caespititius (Lamiaceae) genotypes were performed. Genomic exon–intron structure was also determined for both terpene synthase genes, revealing an organization with seven exons and six introns. The cDNA of Tctps2 was 2,308 bp long and had an open reading frame of 1,794 bp encoding for a protein with 598 amino acids. Tctps5 was longer, mainly due to intron sequences, and presented high intraspecific variability on the plants analyzed. It encoded for a protein of 602 amino acids from an open reading frame of 1,806 bp comprising a total of 2,507 bp genomic sequence. The amino acid sequence of these two active Tctps genes shared 74 % pairwise identity, ranging between 42 and 94 % similarity with about 50 known terpene synthases of other Lamiaceae species. Gene expression revealed a multi-product Tctps2 and Tctps5 enzymes, producing γ-terpinene and α-terpineol as major components, respectively. These enzymatic results were consistent with the monoterpene profile present in T. caespititius field plants, suggesting a transcriptional regulation in leaves. Herewith reported for the first time for this species, these two newly characterized Tctps genes improve the understanding of the molecular mechanisms of reaction responsible for terpene biosynthesis and chemical diversity found in T. caespititius.

Keywords

Gene expression Genomic characterization Terpene synthase α-Terpineol γ-Terpinene Thymus 

Abbreviations

DTT

Dithiothreitol

GPP

Geranyl diphosphate

FPP

Farnesyl diphosphate

IPTG

Isopropyl β-d-1-thiogalactopyranoside

ISSR

Inter-simple sequence repeat

PMSF

Phenylmethylsulfonyl fluoride

RAPD

Random amplification of polymorphic DNA

SPME

Solid phase micro extraction

Tctps2

T. caespititius terpene synthase 2 gene (γ-terpinene synthase)

Tctps5

T. caespititius terpene synthase 5 gene (α-terpineol synthase)

TPS

Terpene synthase

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Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • A. Sofia Lima
    • 1
  • Jette Schimmel
    • 2
  • Brigitte Lukas
    • 3
  • Johannes Novak
    • 3
  • José G. Barroso
    • 1
  • A. Cristina Figueiredo
    • 1
  • Luis G. Pedro
    • 1
  • Jörg Degenhardt
    • 2
  • Helena Trindade
    • 1
  1. 1.Departmento de Biologia Vegetal, Faculdade de Ciências, Centro de Biotecnologia Vegetal, Instituto de Biotecnologia e BioengenhariaUniversidade de LisboaLisbonPortugal
  2. 2.Institute for Pharmacy, AG Pharmaceutical BiotechnologyMartin Luther University Halle-WittenbergHalle (Saale)Germany
  3. 3.Institute for Applied Botany and PharmacognosyUniversity of Veterinary MedicineWienAustria

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