Chrysanthemum expressing a linalool synthase gene ‘smells good’, but ‘tastes bad’to western flower thrips
Ting Yang, Ting ; Stoopen, G.M. ; Thoen, H.P.M. ; Wiegers, G.L. ; Jongsma, M.A. - \ 2013
Plant Biotechnology Journal 11 (2013)7. - ISSN 1467-7644 - p. 875 - 882.
induced plant volatiles - terpenoid pathway - mass-spectrometry - herbivore enemies - tomato fruits - s-linalool - host-plant - emissions - biosynthesis - defense
Herbivore-induced plant volatiles are often involved in direct and indirect plant defence against herbivores. Linalool is a common floral scent and found to be released from leaves by many plants after herbivore attack. In this study, a linalool/nerolidol synthase, FaNES1, was overexpressed in the plastids of chrysanthemum plants (Chrysanthemum morifolium). The volatiles of FaNES1 chrysanthemum leaves were strongly dominated by linalool, but they also emitted small amount of the C11-homoterpene, (3E)-4,8-dimethyl-1,3,7-nonatriene, a derivative of nerolidol. Four nonvolatile linalool glycosides in methanolic extracts were found to be significantly increased in the leaves of FaNES1 plants compared to wild-type plants. They were putatively identified by LC-MS-MS as two linalool–malonyl–hexoses, a linalool–pentose–hexose and a glycoside of hydroxy–linalool. A leaf-disc dual-choice assay with western flower thrips (WFT, Frankliniella occidentalis) showed, initially during the first 15 min of WFT release, that FaNES1 plants were significantly preferred. This gradually reversed into significant preference for the control, however, at 20–28 h after WFT release. The initial preference was shown to be based on the linalool odour of FaNES1 plants by olfactory dual-choice assays using paper discs emitting pure linalool at similar rates as leaf discs. The reversal of preference into deterrence could be explained by the initial nonvolatile composition of the FaNES1 plants, as methanolic extracts were less preferred by WFT. Considering the common occurrence of linalool and its glycosides in plant tissues, it suggests that plants may balance attractive fragrance with ‘poor taste’ using the same precursor compound.
Volatile science? Metabolic engineering of terpenoids in plants
Aharoni, A. ; Jongsma, M.A. ; Bouwmeester, H.J. - \ 2005
Trends in Plant Science 10 (2005)12. - ISSN 1360-1385 - p. 594 - 602.
transgenic arabidopsis plants - linalool synthase gene - isoprenoid biosynthesis - essential oil - monoterpene biosynthesis - diphosphate synthase - s-linalool - functional expression - menthofuran synthase - plastidial pathways
Terpenoids are important for plant survival and also possess biological properties that are beneficial to humans. Here, we describe the state of the art in terpenoid metabolic engineering, showing that significant progress has been made over the past few years. Subcellular targeting of enzymes has demonstrated that terpenoid precursors in subcellular compartments are not as strictly separated as previously thought and that multistep pathway engineering is feasible, even across cell compartments. These engineered plants show that insect behavior is influenced by terpenoids. In the future, we expect rapid progress in the engineering of terpenoid production in plants. In addition to commercial applications, such transgenic plants should increase our understanding of the biological relevance of these volatile secondary metabolites
Metabolic engineering of monoterpende biosysnthesis: two step production of (+)-trans-Isopiperitenol by tobacco
Lücker, J. ; Schwab, W. ; Franssen, M.C.R. ; Plas, L.H.W. van der; Bouwmeester, H.J. ; Verhoeven, H.A. - \ 2004
The Plant Journal 39 (2004)1. - ISSN 0960-7412 - p. 135 - 145.
peppermint mentha-piperita - functional expression - limonene enantiomers - linalool synthase - cdna isolation - s-linalool - plants - mint - (-)-limonene - cloning
Monoterpenoid biosynthesis in tobacco was modified by introducing two subsequent enzymatic activities targeted to different cell compartments. A limonene-3-hydroxylase (lim3h) cDNA was isolated from Mentha spicata L. 'Crispa'. This cDNA was used to re-transform a transgenic Nicotiana tabacum'Petit Havana' SR1 (tobacco) line expressing three Citrus limon L. Burm. f. (lemon) monoterpene synthases producing (+)-limonene, gamma-terpinene and (-)-beta-pinene as their main products. The targeting sequences of these synthases indicate that they are probably localized in the plastids, whereas the sequence information of the P450 hydroxylase indicates targeting to the endoplasmatic reticulum. Despite the different location of the enzymes, the introduced P450 hydroxylase proved to be functional in the transgenic plants as it hydroxylated (+)-limonene, resulting in the emission of (+)-trans-isopiperitenol. Some further modifications of the (+)-trans-isopiperitenol were also detected, resulting in the additional emission of 1,3,8-p-menthatriene, 1,5,8-p-menthatriene, p-cymene and isopiperitenone.
Terpenoid Metabolism in Wild-Type and Transgenic Arabidopsis Plants
Aharoni, A. ; Giri, A.P. ; Deuerlein, S. ; Griepink, F.C. ; Kogel, W.J. de; Verstappen, F.W.A. ; Verhoeven, H.A. ; Jongsma, M.A. ; Schwab, W. ; Bouwmeester, H.J. - \ 2003
The Plant Cell 15 (2003)12. - ISSN 1040-4651 - p. 2866 - 2884.
sesquiterpene cyclase gene - linalool synthase gene - functional expression - monoterpene biosynthesis - isoprenoid biosynthesis - diphosphate synthase - cdna isolation - s-linalool - glandular trichomes - thaliana
Volatile components, such as terpenoids, are emitted from aerial parts of plants and play a major role in the interaction between plants and their environment. Analysis of the composition and emission pattern of volatiles in the model plant Arabidopsis showed that a range of volatile components are released, primarily from flowers. Most of the volatiles detected were monoterpenes and sesquiterpenes, which in contrast to other volatiles showed a diurnal emission pattern. The active terpenoid metabolism in wild-type Arabidopsis provoked us to conduct an additional set of experiments in which transgenic Arabidopsis overexpressing two different terpene synthases were generated. Leaves of transgenic plants constitutively expressing a dual linalool/nerolidol synthase in the plastids (FaNES1) produced linalool and its glycosylated and hydroxylated derivatives. The sum of glycosylated components was in some of the transgenic lines up to 40- to 60-fold higher than the sum of the corresponding free alcohols. Surprisingly, we also detected the production and emission of nerolidol, albeit at a low level, suggesting that a small pool of its precursor farnesyl diphosphate is present in the plastids. Transgenic lines with strong transgene expression showed growth retardation, possibly as a result of the depletion of isoprenoid precursors in the plastids. In dual-choice assays with Myzus persicae, the FaNES1-expressing lines significantly repelled the aphids. Overexpression of a typical cytosolic sesquiterpene synthase resulted in the production of only trace amounts of the expected sesquiterpene, suggesting tight control of the cytosolic pool of farnesyl diphosphate, the precursor for sesquiterpenoid biosynthesis. This study further demonstrates the value of Arabidopsis for studies of the biosynthesis and ecological role of terpenoids and provides new insights into their metabolism in wild-type and transgenic plants
The influence of monoterpene synthase transformation on the odour of tabacco.
Tamer, M.K. el; Smeets, M.A.M. ; Holthuysen, N.T.E. ; Lucker, J. ; Tang, A. ; Roozen, J.P. ; Bouwmeester, H.J. ; Voragen, A.G.J. - \ 2003
Journal of Biotechnology 106 (2003)1. - ISSN 0168-1656 - p. 15 - 21.
linalool synthase - gene-expression - s-linalool - biosynthesis - plants - cancer
Monoterpenes are an important class of terpenoids that are commonly present in plant essential oils. These can be extracted from plants and are used in the flavouring and perfumery industry. Monoterpene synthases are the key enzymes in monoterpene biosynthesis, as they catalyse the cyclisation of the ubiquitous geranyl diphosphate (GDP) to the specific monoterpene skeletons. Tobacco is one of the most studied model plants, it can easily and efficiently be transformed, and is a suitable model to study the release of plant volatiles. Thus, we have isolated monoterpene synthases from lemon, transformed tobacco with these cDNAs and have used human panelists to study the change in fragrance of the transgenic in comparison to the wild type plants. In a triangle test, we found that subjects were capable of smelling significant differences between leaf samples. However, as a result of variability in panel ratings, no significant difference between two sets of transgenic flowers and the wild type tobacco flowers was found for the generated attributes in a descriptive test.