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

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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.

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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)

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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).

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Authors and Affiliations

  1. Université de Lyon, 42023, Saint-Étienne, France

    Frédéric Jullien, Sandrine Moja, Aurélie Bony, Sylvain Legrand, Cécile Petit, Tarek Benabdelkader, Kévin Poirot, Florence Nicolè, Sylvie Baudino & Jean-Louis Magnard

  2. Université de Saint-Etienne, Jean Monnet, 42000, Saint-Étienne, France

    Frédéric Jullien, Sandrine Moja, Aurélie Bony, Cécile Petit, Florence Nicolè, Sylvie Baudino & Jean-Louis Magnard

  3. Laboratoire de Biotechnologies Végétales Appliquées aux Plantes Aromatiques et Médicinales, EA 3061, 23 rue du Dr Michelon, 42000, Saint-Étienne, France

    Frédéric Jullien, Sandrine Moja, Aurélie Bony, Cécile Petit, Florence Nicolè, Sylvie Baudino & Jean-Louis Magnard

  4. Laboratoire Stress Abiotique et Différenciation des végétaux Cultivés, UMR Lille1/INRA 1281, Université Lille 1, 59655, Villeneuve d’Ascq Cedex, France

    Yann Guitton

  5. Laboratoire d’Ecophysiologie Végétale, Ecole Normale Supérieure, Kouba, 16050, Alger, Algeria

    Tarek Benabdelkader

  6. Institut de Chimie de Nice, UMR-CNRS 7272, Faculté des Sciences, Université de Nice-Sophia Antipolis, 06108, Nice Cedex 2, France

    Sébastien Fiorucci

  7. Stress Abiotique et Différenciation des végétaux Cultivés (SADV), UMR INRA 1281, Université Lille Nord de France, Université Lille 1, SN2, 59650, Villeneuve d’Ascq, France

    Sylvain Legrand

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  1. Frédéric Jullien
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  2. Sandrine Moja
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  3. Aurélie Bony
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  4. Sylvain Legrand
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  5. Cécile Petit
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  6. Tarek Benabdelkader
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  7. Kévin Poirot
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  9. Yann Guitton
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  10. Florence Nicolè
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  11. Sylvie Baudino
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  12. Jean-Louis Magnard
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Corresponding author

Correspondence to Frédéric Jullien.

Additional information

Accession numbers: LaCADS: JX401282; LaCARS: JX401283; LaGERDS: JX401284.

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Jullien, F., Moja, S., Bony, A. et al. Isolation and functional characterization of a τ-cadinol synthase, a new sesquiterpene synthase from Lavandula angustifolia . Plant Mol Biol 84, 227–241 (2014). https://doi.org/10.1007/s11103-013-0131-3

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