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Monoterpene engineering in a woody plant Eucalyptus camaldulensis using a limonene synthase cDNA

Kazuaki Ohara

Laboratory of Plant Gene Expression, Research Institute for Sustainable Humanosphere, Kyoto University, Kyoto, Japan

Present address: Central Laboratories for Frontier Technology, Kirin Holdings Co., Ltd., 1‐13‐5 Fukuura, Kanazawa‐ku, Yokohama‐shi, Kanagawa 236‐0004, Japan.Search for more papers by this author
Etsuko Matsunaga

Forestry Science Research Laboratory, Nippon Paper Industries Co. Ltd., Tokyo, Japan

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Kazuya Nanto

Forestry Science Research Laboratory, Nippon Paper Industries Co. Ltd., Tokyo, Japan

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Kyoko Yamamoto

Laboratory of Plant Gene Expression, Research Institute for Sustainable Humanosphere, Kyoto University, Kyoto, Japan

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Kanako Sasaki

Laboratory of Plant Gene Expression, Research Institute for Sustainable Humanosphere, Kyoto University, Kyoto, Japan

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Hiroyasu Ebinuma

Forestry Science Research Laboratory, Nippon Paper Industries Co. Ltd., Tokyo, Japan

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Kazufumi Yazaki

Corresponding Author

Laboratory of Plant Gene Expression, Research Institute for Sustainable Humanosphere, Kyoto University, Kyoto, Japan

*(fax +81 774 38 3623; e‐mail

yazaki@rish.kyoto‐u.ac.jp

)
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First published: 09 December 2009
Cited by: 9

Summary

Metabolic engineering aimed at monoterpene production has become an intensive research topic in recent years, although most studies have been limited to herbal plants including model plants such as Arabidopsis. The genus Eucalyptus includes commercially important woody plants in terms of essential oil production and the pulp industry. This study attempted to modify the production of monoterpenes, which are major components of Eucalyptus essential oil, by introducing two expression constructs containing Perilla frutescens limonene synthase (PFLS) cDNA, whose gene products were designed to be localized in either the plastid or cytosol, into Eucalyptus camaldulensis. The expression of the plastid‐type and cytosol‐type PFLS cDNA in transgenic E. camaldulensis was confirmed by real‐time polymerase chain reaction (PCR). Gas chromatography with a flame ionization detector analyses of leaf extracts revealed that the plastidic and cytosolic expression of PFLS yielded 2.6‐ and 4.5‐times more limonene than that accumulated in wild‐type E. camaldulensis, respectively, while the ectopic expression of PFLS had only a small effect on the emission of limonene from the leaves of E. camaldulensis. Surprisingly, the high level of PFLS in Eucalyptus was accompanied by a synergistic increase in the production of 1,8‐cineole and α‐pinene, two major components of Eucalyptus monoterpenes. This genetic engineering of monoterpenes demonstrated a new potential for molecular breeding in woody plants.

Number of times cited: 9

  • , Tissue-specific production of limonene in Camelina sativa with the Arabidopsis promoters of genes BANYULS and FRUITFULL, Planta, 243, 2, (549), (2016).
  • , Production of mono- and sesquiterpenes in Camelina sativa oilseed, Planta, 242, 3, (693), (2015).
  • , Differential expression of limonene synthase gene affects production and composition of essential oils in leaf and floret of transgenic lavandin (Lavandula × intermedia Emeric ex Loisel.), Plant Biotechnology Reports, 8, 2, (193), (2014).
  • , Metabolic engineering of plant monoterpenes, sesquiterpenes and diterpenes—current status and future opportunities, Plant Biotechnology Journal, 11, 2, (169-196), (2012).
  • , Seasonal variations in chemical composition and fumigant activity of five Eucalyptus essential oils against three moth pests of stored dates in Tunisia, Journal of Stored Products Research, 48, (61), (2012).
  • , Agrobacterium-mediated transformation of Eucalyptus globulus using explants with shoot apex with introduction of bacterial choline oxidase gene to enhance salt tolerance, Plant Cell Reports, 31, 1, (225), (2012).
  • , Comparison of variation in adaptive traits between wild-type and transgenic silver birch (Betula pendula) in a field trial, Tree Genetics & Genomes, 7, 5, (955), (2011).
  • , Metabolic engineering of essential oil in Eucalyptus using Perilla limonene synthase cDNA, Journal of Japan Association on Odor Environment, 42, 4, (248), (2011).
  • , Metabolic engineering for the production of prenylated polyphenols in transgenic legume plants using bacterial and plant prenyltransferases, Metabolic Engineering, 13, 6, (629), (2011).