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Plant Molecular Biology

, Volume 84, Issue 1–2, pp 227–241 | Cite as

Isolation and functional characterization of a τ-cadinol synthase, a new sesquiterpene synthase from Lavandula angustifolia

  • Frédéric Jullien
  • Sandrine Moja
  • Aurélie Bony
  • Sylvain Legrand
  • Cécile Petit
  • Tarek Benabdelkader
  • Kévin Poirot
  • Sébastien Fiorucci
  • Yann Guitton
  • Florence Nicolè
  • Sylvie Baudino
  • Jean-Louis Magnard
Article

Abstract

In this paper we characterize three sTPSs: a germacrene D (LaGERDS), a (E)-β-caryophyllene (LaCARS) and a τ-cadinol synthase (LaCADS). τ-cadinol synthase is reported here for the first time and its activity was studied in several biological models including transiently or stably transformed tobacco species. Three dimensional structure models of LaCADS and Ocimum basilicum γ-cadinene synthase were built by homology modeling using the template structure of Gossypium arboreum δ-cadinene synthase. The depiction of their active site organization provides evidence of the global influence of the enzymes on the formation of τ-cadinol: instead of a unique amino-acid, the electrostatic properties and solvent accessibility of the whole active site in LaCADS may explain the stabilization of the cadinyl cation intermediate. Quantitative PCR performed from leaves and inflorescences showed two patterns of expression. LaGERDS and LaCARS were mainly expressed during early stages of flower development and, at these stages, transcript levels paralleled the accumulation of the corresponding terpene products (germacrene D and (E)-β-caryophyllene). By contrast, the expression level of LaCADS was constant in leaves and flowers. Phylogenetic analysis provided informative results on potential duplication process leading to sTPS diversification in lavender.

Keywords

Lavandulaangustifolia L. 454 Pyrosequencing Terpene synthase Transcript regulation 

Abbreviations

CADS

Cadinol synthase

CARS

β-Caryophyllene synthase

EO(s)

Essential oil(s)

EST(s)

Expressed sequence tag(s)

IPTG

Isopropyl-β-d-thiogalactopyranoside

FPP

Farnesyl diphosphate

GERDS

Germacrene D synthase

GPP

Geranyl diphosphate

LaCADS

L. angustifolia cadinol synthase

LaCARS

L. angustifolia β-caryophyllene synthase

LaGERDS

L. angustifolia germacrene D synthase

mTPS(s)

Monoterpene synthase(s)

Qpcr

Quantitative realtime polymerase chain reaction

sTPS(s)

Sesquiterpene synthase(s)

TPS(s)

Terpene synthase(s)

Notes

Acknowledgments

This work was supported by a Grant of the Rhône-Alpes county. We thank Dr. Busso who kindly provided the pHGGWA vector. We also thank Florence Gros for her technical contribution to this work. The IFF company and CRIEPPAM are also thanked for providing respectively basil rich τ-cadinol and L. angustifolia cv’Diva’ essential oils used as standards for τ-cadinol identification. We also thank Dr. Bendahmane (ENS Lyon) for critical reading and Frédéric Hache and Neil Finn (Department of Foreign Languages, UJM) for text revision).

Supplementary material

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Supplementary material 1 (PPTX 24219 kb)
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Supplementary material 2 (DOCX 85 kb)
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Supplementary material 3 (PPTX 67 kb)
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Supplementary material 4 (PPTX 1059 kb)

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Copyright information

© Springer Science+Business Media Dordrecht 2013

Authors and Affiliations

  • Frédéric Jullien
    • 1
    • 2
    • 3
  • Sandrine Moja
    • 1
    • 2
    • 3
  • Aurélie Bony
    • 1
    • 2
    • 3
  • Sylvain Legrand
    • 1
    • 7
  • Cécile Petit
    • 1
    • 2
    • 3
  • Tarek Benabdelkader
    • 1
    • 5
  • Kévin Poirot
    • 1
  • Sébastien Fiorucci
    • 6
  • Yann Guitton
    • 4
  • Florence Nicolè
    • 1
    • 2
    • 3
  • Sylvie Baudino
    • 1
    • 2
    • 3
  • Jean-Louis Magnard
    • 1
    • 2
    • 3
  1. 1.Université de LyonSaint-ÉtienneFrance
  2. 2.Université de Saint-Etienne, Jean MonnetSaint-ÉtienneFrance
  3. 3.Laboratoire de Biotechnologies Végétales Appliquées aux Plantes Aromatiques et MédicinalesSaint-ÉtienneFrance
  4. 4.Laboratoire Stress Abiotique et Différenciation des végétaux Cultivés, UMR Lille1/INRA 1281Université Lille 1Villeneuve d’Ascq CedexFrance
  5. 5.Laboratoire d’Ecophysiologie VégétaleEcole Normale SupérieureAlgerAlgeria
  6. 6.Institut de Chimie de Nice, UMR-CNRS 7272, Faculté des SciencesUniversité de Nice-Sophia AntipolisNice Cedex 2France
  7. 7.Stress Abiotique et Différenciation des végétaux Cultivés (SADV), UMR INRA 1281Université Lille Nord de FranceVilleneuve d’AscqFrance

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