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JXB Advance Access originally published online on June 10, 2009
Journal of Experimental Botany 2009 60(11):3203-3219; doi:10.1093/jxb/erp162
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© 2009 The Author(s).
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. This paper is available online free of all access charges (see http://jxb.oxfordjournals.org/open_access.html for further details)

RESEARCH PAPER

Two terpene synthases are responsible for the major sesquiterpenes emitted from the flowers of kiwifruit (Actinidia deliciosa)

Niels J. Nieuwenhuizen1,*, Mindy Y. Wang1, Adam J. Matich2, Sol A. Green1, Xiuyin Chen1, Yar-Khing Yauk1, Lesley L. Beuning1, Dinesh A. Nagegowda3, Natalia Dudareva3 and Ross G. Atkinson1

1The New Zealand Institute for Plant and Food Research Ltd, Private Bag 92 169, Auckland, New Zealand
2The New Zealand Institute for Plant and Food Research Ltd, Private Bag 11 030, Palmerston Nth, New Zealand
3Department of Horticulture and Landscape Architecture, Purdue University, West Lafayette, IN 47907, USA

* To whom correspondence should be addressed: E-mail: nnieuwenhuizen{at}hortresearch.co.nz

Kiwifruit vines rely on bees for pollen transfer between spatially separated male and female individuals and require synchronized flowering to ensure pollination. Volatile terpene compounds, which are important cues for insect pollinator attraction, were studied by dynamic headspace sampling in the major green-fleshed kiwifruit (Actinidia deliciosa) cultivar ‘Hayward’ and its male pollinator ‘Chieftain’. Terpene volatile levels showed a profile dominated by the sesquiterpenes {alpha}-farnesene and germacrene D. These two compounds were emitted by all floral tissues and could be observed throughout the day, with lower levels at night. The monoterpene (E)-β-ocimene was also detected in flowers but was emitted predominantly during the day and only from petal tissue. Using a functional genomics approach, two terpene synthase (TPS) genes were isolated from a ‘Hayward’ petal EST library. Bacterial expression and transient in planta data combined with analysis by enantioselective gas chromatography revealed that one TPS produced primarily (E,E)-{alpha}-farnesene and small amounts of (E)-β-ocimene, whereas the second TPS produced primarily (+)-germacrene D. Subcellular localization using GFP fusions showed that both enzymes were localized in the cytoplasm, the site for sesquiterpene production. Real-time PCR analysis revealed that both TPS genes were expressed in the same tissues and at the same times as the corresponding floral volatiles. The results indicate that two genes can account for the major floral sesquiterpene volatiles observed in both male and female A. deliciosa flowers.

Key words: Actinidia, {alpha}-farnesene, floral volatiles, germacrene D, kiwifruit, ocimene, terpene, terpene synthases

Received 3 March 2009; Revised 15 April 2009 Accepted 20 April 2009


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The way the dioecious plant Actinidia deliciosa attracts bees: critical role of volatile terpenes released from kiwifruit flowers of both genotypes
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