Publications

• Terpenoid metabolism in wild-type and transgenic Arabidopsis plants.

  Aharoni A., Giri A.P., Deuerlein S., Griepink F., de Kogel W.J., Verstappen F.W., Verhoeven H.A., Jongsma M.A., Schwab W., Bouwmeester H.J.

  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 ar ... >> More

  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. << Less

  Plant Cell 15:2866-2884(2003) [PubMed] [EuropePMC]

• Demonstration and characterization of (E)-nerolidol synthase from maize: a herbivore-inducible terpene synthase participating in (3E)-4,8-dimethyl-1,3,7-nonatriene biosynthesis.

  Degenhardt J., Gershenzon J.

  Upon herbivore attack, maize (Zea mays L.) emits a mixture of volatile compounds that attracts herbivore enemies to the plant. One of the major components of this mixture is an unusual acyclic C11 homoterpene, (3E)-4,8-dimethyl-1,3,7-nonatriene (DMNT), which is also emitted by many other species f ... >> More

  Upon herbivore attack, maize (Zea mays L.) emits a mixture of volatile compounds that attracts herbivore enemies to the plant. One of the major components of this mixture is an unusual acyclic C11 homoterpene, (3E)-4,8-dimethyl-1,3,7-nonatriene (DMNT), which is also emitted by many other species following herbivore damage. Biosynthesis of DMNT has been previously shown to proceed via the sesquiterpene alcohol, (E)-nerolidol. Here we demonstrate an enzyme activity that converts farnesyl diphosphate, the universal precursor of sesquiterpenes, to (3S)-(E)-nerolidol in cell-free extracts of maize leaves that had been fed upon by Spodoptera littoralis. The properties of this (E)-nerolidol synthase resemble those of other terpene synthases. Evidence for its participation in DMNT biosynthesis includes the direct incorporation of deuterium-labeled (E)-nerolidol into DMNT and the close correlation between increases in (E)-nerolidol synthase activity and DMNT emission after herbivore damage. Since farnesyl diphosphate has many other metabolic fates, (E)-nerolidol synthase may represent the first committed step of DMNT biosynthesis in maize. However, the formation of this unusual acyclic terpenoid appears to be regulated at both the level of (E)-nerolidol synthase and at later steps in the pathway. << Less

  Planta 210:815-822(2000) [PubMed] [EuropePMC]

• Nonseed plant Selaginella moellendorfii has both seed plant and microbial types of terpene synthases.

  Li G., Kollner T.G., Yin Y., Jiang Y., Chen H., Xu Y., Gershenzon J., Pichersky E., Chen F.

  Terpene synthases (TPSs) are pivotal enzymes for the biosynthesis of terpenoids, the largest class of secondary metabolites made by plants and other organisms. To understand the basis of the vast diversification of these enzymes in plants, we investigated Selaginella moellendorffi, [corrected] a n ... >> More

  Terpene synthases (TPSs) are pivotal enzymes for the biosynthesis of terpenoids, the largest class of secondary metabolites made by plants and other organisms. To understand the basis of the vast diversification of these enzymes in plants, we investigated Selaginella moellendorffi, [corrected] a nonseed vascular plant. The genome of this species was found to contain two distinct types of TPS genes. The first type of genes, which was designated as S. moellendorffi [corrected] TPS genes (SmTPSs), consists of 18 members. SmTPSs share common ancestry with typical seed plant TPSs. Selected members of the SmTPSs were shown to encode diterpene synthases. The second type of genes, designated as S. moellendorffi [corrected] microbial TPS-like genes (SmMTPSLs), consists of 48 members. Phylogenetic analysis showed that SmMTPSLs are more closely related to microbial TPSs than other plant TPSs. Selected SmMTPSLs were determined to function as monoterpene and sesquiterpene synthases. Most of the products formed were typical monoterpenes and sesquiterpenes that have been previously shown to be synthesized by classical plant TPS enzymes. Some in vitro products of the characterized SmMTPSLs were detected in the headspace of S. moellendorffi [corrected] plants treated with the fungal elicitor alamethicin, showing that they are also formed in the intact plant. The presence of two distinct types of TPSs in the genome of S. moellendorffi [corrected] raises the possibility that the TPSs in other plant species may also have more than one evolutionary origin. << Less

  Proc. Natl. Acad. Sci. U.S.A. 109:14711-14715(2012) [PubMed] [EuropePMC]

  This publication is cited by 4 other entries.

• Gain and loss of fruit flavor compounds produced by wild and cultivated strawberry species.

  Aharoni A., Giri A.P., Verstappen F.W., Bertea C.M., Sevenier R., Sun Z., Jongsma M.A., Schwab W., Bouwmeester H.J.

  The blends of flavor compounds produced by fruits serve as biological perfumes used to attract living creatures, including humans. They include hundreds of metabolites and vary in their characteristic fruit flavor composition. The molecular mechanisms by which fruit flavor and aroma compounds are ... >> More

  The blends of flavor compounds produced by fruits serve as biological perfumes used to attract living creatures, including humans. They include hundreds of metabolites and vary in their characteristic fruit flavor composition. The molecular mechanisms by which fruit flavor and aroma compounds are gained and lost during evolution and domestication are largely unknown. Here, we report on processes that may have been responsible for the evolution of diversity in strawberry (Fragaria spp) fruit flavor components. Whereas the terpenoid profile of cultivated strawberry species is dominated by the monoterpene linalool and the sesquiterpene nerolidol, fruit of wild strawberry species emit mainly olefinic monoterpenes and myrtenyl acetate, which are not found in the cultivated species. We used cDNA microarray analysis to identify the F. ananassa Nerolidol Synthase1 (FaNES1) gene in cultivated strawberry and showed that the recombinant FaNES1 enzyme produced in Escherichia coli cells is capable of generating both linalool and nerolidol when supplied with geranyl diphosphate (GPP) or farnesyl diphosphate (FPP), respectively. Characterization of additional genes that are very similar to FaNES1 from both the wild and cultivated strawberry species (FaNES2 and F. vesca NES1) showed that only FaNES1 is exclusively present and highly expressed in the fruit of cultivated (octaploid) varieties. It encodes a protein truncated at its N terminus. Green fluorescent protein localization experiments suggest that a change in subcellular localization led to the FaNES1 enzyme encountering both GPP and FPP, allowing it to produce linalool and nerolidol. Conversely, an insertional mutation affected the expression of a terpene synthase gene that differs from that in the cultivated species (termed F. ananassa Pinene Synthase). It encodes an enzyme capable of catalyzing the biosynthesis of the typical wild species monoterpenes, such as alpha-pinene and beta-myrcene, and caused the loss of these compounds in the cultivated strawberries. The loss of alpha-pinene also further influenced the fruit flavor profile because it was no longer available as a substrate for the production of the downstream compounds myrtenol and myrtenyl acetate. This phenomenon was demonstrated by cloning and characterizing a cytochrome P450 gene (Pinene Hydroxylase) that encodes the enzyme catalyzing the C10 hydroxylation of alpha-pinene to myrtenol. The findings shed light on the molecular evolutionary mechanisms resulting in different flavor profiles that are eventually selected for in domesticated species. << Less

  Plant Cell 16:3110-3131(2004) [PubMed] [EuropePMC]

  This publication is cited by 1 other entry.

• Herbivore-induced terpenoid emission in Medicago truncatula: concerted action of jasmonate, ethylene and calcium signaling.

  Arimura G., Garms S., Maffei M., Bossi S., Schulze B., Leitner M., Mithofer A., Boland W.

  Plant volatiles emitted by Medicago truncatula in response to feeding larvae of Spodoptera exigua are composed of a complex blend of terpenoids. The cDNAs of three terpene synthases (TPSs), which contribute to the blend of terpenoids, were cloned from M. truncatula. Their functional characterizati ... >> More

  Plant volatiles emitted by Medicago truncatula in response to feeding larvae of Spodoptera exigua are composed of a complex blend of terpenoids. The cDNAs of three terpene synthases (TPSs), which contribute to the blend of terpenoids, were cloned from M. truncatula. Their functional characterization proved MtTPS1 to be a beta-caryophyllene synthase and MtTPS5 to be a multi-product sesquiterpene synthase. MtTPS3 encodes a bifunctional enzyme producing (E)-nerolidol and geranyllinalool (precursors of C11 and C16 homoterpenes) from different prenyl diphosphates serving as substrates. The addition of jasmonic acid (JA) induced expression of the TPS genes, but terpenoid emission was higher from plants treated with JA and the ethylene precursor 1-amino-cyclopropyl-1-carboxylic acid. Compared to infested wild-type M. truncatula plants, lower amounts of various sesquiterpenes and a C11-homoterpene were released from an ethylene-insensitive mutant skl. This difference coincided with lower transcript levels of MtTPS5 and of 1-deoxy-D: -xylulose-5-phosphate synthase (MtDXS2) in the damaged skl leaves. Moreover, ethephon, an ethylene-releasing compound, modified the extent and mode of the herbivore-stimulated Ca2+ variations in the cytoplasm that is necessary for both JA and terpene biosynthesis. Thus, ethylene contributes to the herbivory-induced terpenoid biosynthesis at least twice: by modulating both early signaling events such as cytoplasmic Ca2+-influx and the downstream JA-dependent biosynthesis of terpenoids. << Less

  Planta 227:453-464(2008) [PubMed] [EuropePMC]

• Spider mite-induced (3S)-(E)-nerolidol synthase activity in cucumber and lima bean. The first dedicated step in acyclic C11-homoterpene biosynthesis.

  Bouwmeester H.J., Verstappen F.W., Posthumus M.A., Dicke M.

  Many plant species respond to herbivory with de novo production of a mixture of volatiles that attracts carnivorous enemies of the herbivores. One of the major components in the blend of volatiles produced by many different plant species in response to herbivory by insects and spider mites is the ... >> More

  Many plant species respond to herbivory with de novo production of a mixture of volatiles that attracts carnivorous enemies of the herbivores. One of the major components in the blend of volatiles produced by many different plant species in response to herbivory by insects and spider mites is the homoterpene 4,8-dimethyl-1,3(E), 7-nonatriene. One study (J. Donath, W. Boland [1995] Phytochemistry 39: 785-790) demonstrated that a number of plant species can convert the acyclic sesquiterpene alcohol (3S)-(E)-nerolidol to this homoterpene. Cucumber (Cucumis sativus L.) and lima bean (Phaseolus lunatus L.) both produce 4,8-dimethyl-1,3(E),7-nonatriene in response to herbivory. We report the presence in cucumber and lima bean of a sesquiterpene synthase catalyzing the formation of (3S)-(E)-nerolidol from farnesyl diphosphate. The enzyme is inactive in uninfested cucumber leaves, slightly active in uninfested lima bean leaves, and strongly induced by feeding of the two-spotted spider mite (Tetranychus urticae Koch) on both plant species, but not by mechanical wounding. The activities of the (3S)-(E)-nerolidol synthase correlated well with the levels of release of 4, 8-dimethyl-1,3(E),7-nonatriene from the leaves of the different treatments. Thus, (3S)-(E)-nerolidol synthase is a good candidate for a regulatory role in the release of the important signaling molecule 4,8-dimethyl-1,3(E),7-nonatriene. << Less

  Plant Physiol 121:173-180(1999) [PubMed] [EuropePMC]