Register      Login
Functional Plant Biology Functional Plant Biology Society
Plant function and evolutionary biology
RESEARCH ARTICLE

Characterisation of an (S)-linalool synthase from kiwifruit (Actinidia arguta) that catalyses the first committed step in the production of floral lilac compounds

Xiuyin Chen A C , Yar-Khing Yauk A C , Niels J. Nieuwenhuizen A , Adam J. Matich B , Mindy Y. Wang A , Ramon Lopez Perez A , Ross G. Atkinson A D and Lesley L. Beuning A
+ Author Affiliations
- Author Affiliations

A The New Zealand Institute for Plant and Food Research Limited (PFR), Private Bag 92 169, Auckland 1142, New Zealand.

B PFR, Private Bag 11600, Palmerston North 4442, New Zealand.

C These authors contributed equally to this work.

D Corresponding author. Email: ross.atkinson@plantandfood.co.nz

Functional Plant Biology 37(3) 232-243 https://doi.org/10.1071/FP09179
Submitted: 17 July 2009  Accepted: 6 November 2009   Published: 25 February 2010

Abstract

Kiwifruit (Actinidia spp. Lindl.) flowers and fruit contain many compounds of interest to the flavour and fragrance industries. In particular, Actinidia arguta (Sieb. et Zucc.) Planch. ex Miq. flowers produce β-linalool and important derivatives thereof, including linalool oxides, lilac aldehydes, alcohols and alcohol epoxides. Dynamic headspace sampling of whole A. arguta flowers showed that the peak emission rate of linalool, lilac alcohols and lilac aldehydes occurred around 0800 hours. After solvent extraction, linalool levels remained constant throughout the day and night, but lilac alcohol levels peaked at noon. In whole flowers, linalool was found predominantly in pistils and petals, and the lilac compounds were found mainly in petals. Two highly homologous (96.6% nucleotide identity) terpene synthase cDNA sequences, AaLS1 and ApLS1, were isolated from A. arguta and Actinidia polygama (Sieb. et Zucc.) Maxim flower EST libraries respectively. Real-time PCR analysis revealed that AaLS1 was expressed constitutively throughout the day and night, and primarily in petal tissue. Functional analysis in Escherichia coli showed that AaLS1 and ApLS1 each encoded a linalool synthase which was confirmed by transient expression in planta. Enantioselective gas chromatography revealed that both terpene synthases produced only (S)-(+)-linalool. AaLS1, therefore, is likely to be the key enzyme producing the (S)-linalool precursor of the lilac alcohols and aldehydes in A. arguta flowers.

Additional keywords: flower, terpene, volatile.


Acknowledgements

We thank Andrew Kralicek and Catrin Guenther for critically reviewing the manuscript, Mark McNeilage and Robert Winz for help with flower collections and Elspeth McRae, Miva Splawinski and Ellen Friel for contributing to preliminary AaLS1 studies. This work was funded by the New Zealand Foundation for Research Science and Technology (C06X0403).


References


Aharoni A, Giri AP, Deuerlein S, Griepink F, de Kogel WJ, Verstappen FW, Verhoeven HA, Jongsma MA, Schwab W, Bouwmeester HJ (2003) Terpenoid metabolism in wild-type and transgenic Arabidopsis plants. The Plant Cell 15, 2866–2884.
Crossref | GoogleScholarGoogle Scholar | PubMed | CAS | [Verified 8 January 2010]

Nieuwenhuizen NJ, Beuning LL, Sutherland PW, Sharma NN, Cooney JM, Bieleski LRF, Schröder R, MacRae EA, Atkinson RG (2007) Identification and characterisation of acidic and novel basic forms of actinidin, the highly abundant cysteine protease from kiwifruit. Functional Plant Biology 34, 946–961.
Crossref | GoogleScholarGoogle Scholar | CAS | open url image1

Nieuwenhuizen NJ, Wang MY, Matich AJ, Green SA, Chen X, Yauk Y-K, Beuning LL, Nagegowda DA, Dudareva N, Atkinson RG (2009) Two terpene synthases are responsible for the major terpenes emitted from the flowers of kiwifruit (Actinidia deliciosa). Journal of Experimental Botany 60, 3203–3219.
Crossref | GoogleScholarGoogle Scholar | PubMed | CAS | open url image1

Pichersky E, Lewinsohn E, Croteau R (1995) Purification and characterization of S-linalool synthase, an enzyme involved in the production of floral scent in Clarkia breweri. Archives of Biochemistry and Biophysics 316, 803–807.
Crossref | GoogleScholarGoogle Scholar | PubMed | CAS | open url image1

Reinhard J, Srinivasan MV, Zhang S (2004) Olfaction: scent-triggered navigation in honeybees. Nature 427, 411.
Crossref | GoogleScholarGoogle Scholar | PubMed | CAS | open url image1

Sitrit Y, Ninio R, Bar E, Golan E, Larkov O, Ravid U, Lewinsohn E (2004) S-linalool synthase activity in developing fruit of the columnar cactus koubo (Cereus peruvianus (L.) Miller). Plant Science 167, 1257–1262.
Crossref | GoogleScholarGoogle Scholar | CAS | open url image1

Soria AC, Gonzalez M, de Lorenzo C, Martinez-Castro I, Sanz J (2004) Characterization of artisanal honeys from Madrid (Central Spain) on the basis of their melissopalynological, physicochemical and volatile composition data. Food Chemistry 85, 121–130.
Crossref | GoogleScholarGoogle Scholar | CAS | open url image1

Starks CM, Back K, Chappell J, Noel JP (1997) Structural basis for cyclic terpene biosynthesis by tobacco 5-epi-aristolochene synthase. Science 277, 1815–1820.
Crossref | GoogleScholarGoogle Scholar | PubMed | CAS | open url image1

Steven D (1988) Chinese pollinators identified. New Zealand Kiwifruit November 1988, 15. open url image1

Studier FW (2005) Protein production by auto-induction in high density shaking cultures. Protein Expression and Purification 41, 207–234.
Crossref | GoogleScholarGoogle Scholar | PubMed | CAS | open url image1

Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG (1997) The ClustalX windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Research 25, 4876–4882.
Crossref | GoogleScholarGoogle Scholar | CAS | PubMed | open url image1

Tollsten L, Bergstrom LG (1993) Fragrance chemotypes of Platanthera (Orchidaceae) – the result of adaptation to pollinating moths. Nordic Journal of Botany 13, 607–613.
Crossref | GoogleScholarGoogle Scholar | open url image1

Ulland S, Ian E, Borg-Karlson AK, Mustaparta H (2006) Discrimination between enantiomers of linalool by olfactory receptor neurons in the cabbage moth Mamestra brassicae (L.). Chemical Senses 31, 325–334.
Crossref | GoogleScholarGoogle Scholar | PubMed | CAS | open url image1

van Schie CCN, Haring MA, Schuurink RC (2007) Tomato linalool synthase is induced in trichomes by jasmonic acid. Plant Molecular Biology 64, 251–263.
Crossref | GoogleScholarGoogle Scholar | PubMed | open url image1

Vancanneyt G, Sanz C, Farmaki T, Paneque M, Ortego F, Castanera P, Sanchez-Serrano JJ (2001) Hydroperoxide lyase depletion in transgenic potato plants leads to an increase in aphid performance. Proceedings of the National Academy of Sciences of the United States of America 98, 8139–8144.
Crossref | GoogleScholarGoogle Scholar | PubMed | CAS | open url image1

Wei S, Marton I, Dekel M, Shalitin D, Lewinsohn E, Bravdo BA, Shoseyov O (2004) Manipulating volatile emission in tobacco leaves by expressing Aspergillus niger beta-glucosidase in different subcellular compartments. Plant Biotechnology Journal 2, 341–350.
Crossref | GoogleScholarGoogle Scholar | PubMed | CAS | open url image1

Williams DC, McGarvey DJ, Katahira EJ, Croteau R (1998) Truncation of limonene synthase preprotein provides a fully active ‘pseudomature’ form of this monoterpene cyclase and reveals the function of the amino-terminal arginine pair. Biochemistry 37, 12213–12220.
Crossref | GoogleScholarGoogle Scholar | PubMed | CAS | open url image1

Yuan JS, Kollner TG, Wiggins G, Grant J, Degenhardt J, Chen F (2008) Molecular and genomic basis of volatile-mediated indirect defense against insects in rice. The Plant Journal 55, 491–503.
Crossref | GoogleScholarGoogle Scholar | PubMed | CAS | open url image1