Biosynthesis Of Volatile Terpenes Research Articles

Transcriptome Analyses Reveal the Aroma Terpeniods Biosynthesis Pathways of Primula forbesii Franch. and the Functional Characterization of the PfDXS2 Gene.

Primula forbesii Franch. is a unique biennial herb with a strong floral fragrance, making it an excellent material for studying the aroma characteristics of the genus Primula. The floral scent is an important ornamental trait that facilitates fertilization. However, the molecular mechanism regulating the floral scent in Primula is unknown. In order to better understand the biological mechanisms of floral scents in this species, this study used RNA sequencing analysis to discuss the first transcriptome sequence of four flowering stages of P. forbesii, which generated 12 P. forbesii cDNA libraries with 79.64 Gb of clean data that formed 51,849 unigenes. Moreover, 53.26% of the unigenes were annotated using public databases. P. forbesii contained 44 candidate genes covering all known enzymatic steps for the biosynthesis of volatile terpenes, the major contributor to the flower's scent. Finally, 1-deoxy-d-xylulose 5-phosphate synthase gene of P. forbesii (PfDXS2, MK370094), the first key enzyme gene in the 2-c-methyl-d-erythritol 4-phosphate (MEP) pathway of terpenoids, was cloned and functionally verified using virus-induced gene silencing (VIGs). The results showed that PfDXS2-silencing significantly reduced the relative concentrations of main volatile terpenes. This report is the first to present molecular data related to aroma metabolites biosynthesis pathways and the functional characterization of any P. forbesii gene. The data on RNA sequencing provide comprehensive information for further analysis of other plants of the genus Primula.

The hops (Humulus lupulus) genome contains a mid-sized terpene synthase family that shows wide functional and allelic diversity

BackgroundHops (Humulus lupulus L.) are a dioecious climbing perennial, with the dried mature “cones” (strobili) of the pistillate/female inflorescences being widely used as both a bittering agent and to enhance the flavour of beer. The glandular trichomes of the bract and bracteole flowering structures of the cones produce an abundance of secondary metabolites, such as terpenoids, bitter acids and prenylated phenolics depending on plant genetics, developmental stage and environment. More knowledge is required on the functional and allelic diversity of terpene synthase (TPS) genes responsible for the biosynthesis of volatile terpenes to assist in flavour-directed hop breeding.ResultsMajor volatile terpene compounds were identified using gas chromatography–mass spectrometry (GC–MS) in the ripe cones of twenty-one hop cultivars grown in New Zealand. All cultivars produced the monoterpene β-myrcene and the sesquiterpenes α-humulene and β-caryophyllene, but the quantities varied broadly. Other terpenes were found in large quantities in only a smaller subset of cultivars, e.g. β-farnesene (in seven cultivars) and α-pinene (in four). In four contrasting cultivars (Wakatu™, Wai-iti™, Nelson Sauvin™, and ‘Nugget’), terpene production during cone development was investigated in detail, with concentrations of some of the major terpenes increasing up to 1000-fold during development and reaching maximal levels from 50–60 days after flowering. Utilising the published H. lupulus genome, 87 putative full-length and partial terpene synthase genes were identified. Alleles corresponding to seven TPS genes were amplified from ripe cone cDNA from multiple cultivars and subsequently functionally characterised by transient expression in planta. Alleles of the previously characterised HlSTS1 produced humulene/caryophyllene as the major terpenes. HlRLS alleles produced (R)-(-)-linalool, whilst alleles of two sesquiterpene synthase genes, HlAFS1 and HlAFS2 produced α-farnesene. Alleles of HlMTS1, HlMTS2 and HlTPS1 were inactive in all the hop cultivars studied.ConclusionsAlleles of four TPS genes were identified and shown to produce key aroma volatiles in ripe hop cones. Multiple expressed but inactive TPS alleles were also identified, suggesting that extensive loss-of-function has occurred during domestication and breeding of hops. Our results can be used to develop hop cultivars with novel/improved terpene profiles using marker-assisted breeding strategies to select for, or against, specific TPS alleles.

Identification of QTL controlling volatile terpene contents in tea plant (Camellia sinensis) using a high-aroma 'Huangdan' x 'Jinxuan' F1 population.

Aroma is an important factor affecting the character and quality of tea. The improvement of aroma trait is a crucial research direction of tea plant breeding. Volatile terpenes, as the major contributors to the floral odors of tea products, also play critical roles in the defense responses of plants to multiple stresses. However, previous studies have largely focused on the aroma formation during the manufacture of tea or the comparison of raw tea samples. The mechanisms causing different aroma profiles between tea cultivars have remained underexplored. In the current study, a high-density genetic linkage map of tea plant was constructed based on an F1 population of 'Huangdan' × 'Jinxuan' using genotyping by sequencing. This linkage map covered 1754.57 cM and contained 15 linkage groups with a low inter-marker distance of 0.47 cM. A total of 42 QTLs associated with eight monoterpene contents and 12 QTLs associated with four sesquiterpenes contents were identified with the average PVE of 12.6% and 11.7% respectively. Furthermore, six candidate genes related to volatile terpene contents were found in QTL cluster on chromosome 5 by RNA-seq analysis. This work will enrich our understanding of the molecular mechanism of volatile terpene biosynthesis and provide a theoretical basis for tea plant breeding programs for aroma quality improvement.

The chromosome-scale genome assembly of Jasminum sambac var. unifoliatum provides insights into the formation of floral fragrance

Jasmine [Jasminum sambac (L.) Ait.], a tropical and subtropical plant emits a sweet, heady fragrance during flower opening. However, the molecular mechanisms underlying this phenomenon remain largely unknown. In the present study, integrated Illumina sequencing, Pacbio sequencing, and high-throughput chromatin conformation capture (Hi-C) scaffolding was used to generate a 495.60 Mb genome assembly of J. sambac var. unifoliatum cultivar ‘Fuzhou Single-petal’ (JSU-FSP), with contig N50 of 16.88 Mb; 96.23% of the assembly was assigned to 13 pseudochromosomes. The genome harbors 30 989 protein-coding genes, and 49.47% of the assembled sequences are repetitive sequences. The analysis of duplication modes showed that 51% of genes were duplicated through dispersed duplication, and expanded gene families are mainly involved in photosynthesis, which may be responsible for the light-loving characteristic specific to jasmine. Transcriptome analysis revealed that at least 35 structural genes involved in the biosynthesis of volatile terpenes (VTs), volatile phenylpropanoid/benzenoids (VPBs), fatty acid-derived volatiles (FADVs), and indole were highly expressed in the flower-opening stage, both preharvest and postharvest, and are proposed to be important in endowing flower aroma. Additionally, at least 28 heat shock protein (HSP) and 11 β-glucosidase (BGLU) genes may be involved in the formation of floral fragrance. These findings provide insights into the formation of the floral fragrance of jasmine and will promote germplasm utilization for breeding improved jasmine varieties.

ZmMYC2s play important roles in maize responses to simulated herbivory and jasmonate.

Both herbivory and jasmonic acid (JA) activate the biosynthesis of defensive metabolites in maize, but the mechanism underlying this remains unclear. We generated maize mutants in which ZmMYC2a and ZmMYC2b, two transcription factor genes important in JA signaling, were individually or both knocked out. Genetic and biochemical analyses were used to elucidate the functions of ZmMYC2 proteins in the maize response to simulated herbivory and JA. Compared with the wild-type (WT) maize, the double mutant myc2ab was highly susceptible to insects, and the levels of benzoxazinoids and volatile terpenes, and the levels of their biosynthesis gene transcripts, were much lower in the mutants than in the WT maize after simulated insect feeding or JA treatment. Moreover, ZmMYC2a and ZmMYC2b played a redundant role in maize resistance to insects and JA signaling. Transcriptome and Cleavage Under Targets and Tagmentation-Sequencing (CUT&Tag-Seq) analysis indicated that ZmMYC2s physically targeted 60% of the JA-responsive genes, even though only 33% of these genes were transcriptionally ZmMYC2-dependent. Importantly, CUT&Tag-Seq and dual luciferase assays revealed that ZmMYC2s transactivate the benzoxazinoid and volatile terpene biosynthesis genes IGPS1/3, BX10/11/12/14, and TPS10/2/3/4/5/8 by directly binding to their promoters. Furthermore, several transcription factors physically targeted by ZmMYC2s were identified, and these are likely to function in the regulation of benzoxazinoid biosynthesis. This work reveals the transcriptional regulatory landscapes of both JA signaling and ZmMYC2s in maize and provides comprehensive mechanistic insight into how JA signaling modulates defenses in maize responses to herbivory through ZmMYC2s.

Effect of Developmental Stages on Genes Involved in Middle and Downstream Pathway of Volatile Terpene Biosynthesis in Rose Petals.

Terpenoids are economically and ecologically important compounds, and they are vital constituents in rose flower fragrance and rose essential oil. The terpene synthase genes (TPSs), trans-prenyltransferases genes (TPTs), NUDX1 are involved in middle and downstream pathway of volatile terpene biosynthesis in rose flowers. We identified 7 complete RcTPTs, 49 complete RcTPSs, and 9 RcNUDX1 genes in the genome of Rosa chinensis. During the flower opening process of butterfly rose (Rosa chinensis ‘Mutabilis’, MU), nine RcTPSs expressed in the petals of opening MU flowers exhibited two main expression trends, namely high and low, in old and fresh petals. Five short-chain petal-expressed RcTPTs showed expression patterns corresponding to RcTPSs. Analysis of differential volatile terpenes and differential expressed genes indicated that higher emission of geraniol from old MU petals might be related to the RcGPPS expression. Comprehensive analysis of volatile emission, sequence structure, micro-synteny and gene expression suggested that RcTPS18 may encode (E,E)-α-farnesene synthase. These findings may be useful for elucidating the molecular mechanism of terpenoid metabolism in rose and are vital for future studies on terpene regulation.

The scarecrow-like transcription factor SlSCL3 regulates volatile terpene biosynthesis and glandular trichome size in tomato (Solanum lycopersicum).

Tomato (Solanum lycopersicum L.) type VI glandular trichomes that occur on the surface of leaves, stems, young fruits and flowers produce and store a blend of volatile monoterpenes and sesquiterpenes. These compounds play important roles in the interaction with pathogens and herbivorous insects. Although the function of terpene synthases in the biosynthesis of volatile terpenes in tomato has been comprehensively investigated, the deciphering of their transcriptional regulation is only just emerging. We selected transcription factors that are over-expressed in trichomes based on existing transcriptome data and silenced them individually by virus-induced gene silencing. Of these, SlSCL3, a scarecrow-like (SCL) subfamily transcription factor, led to a significant decrease in volatile terpene content and expression of the corresponding terpene synthase genes when its transcription level was downregulated. Overexpression of SlSCL3 dramatically increased both the volatile terpene content and glandular trichome size, whereas its homozygous mutants showed reduced terpene biosynthesis. However, its heterozygous mutants also showed a significantly elevated volatile terpene content and enlarged glandular trichomes, similar to the overexpression plants. SlSCL3 modulates the expression of terpene biosynthetic pathway genes by transcriptional activation, but neither direct protein-DNA binding nor interaction with known regulators was observed. Moreover, transcript levels of the endogenous copy of SlSCL3 were decreased in the overexpression plants but increased in the heterozygous and homozygous mutants, suggesting feedback repression of its own promoter. Taken together, our results provide new insights into the role of SlSCL3 in the complex regulation of volatile terpene biosynthesis and glandular trichome development in tomato.

Biosynthesis of volatile terpenes that accumulate in the secretory cavities of young leaves of Japanese pepper (Zanthoxylum piperitum): Isolation and functional characterization of monoterpene and sesquiterpene synthase genes.

Volatile terpenes are ones of the characteristic aromas of Japanese pepper (Zanthoxylum piperitum). It has been hypothesized that the specialized epithelial cells surrounding the secretory cavities of Japanese pepper fruits and leaves are responsible for the synthesis of monoterpenes and sesquiterpenes, which are generally produced by terpene synthases (TPSs); however, direct evidence for the formation of terpenes in Japanese pepper remains elusive. Here we report that monoterpenes and sesquiterpenes accumulate inside the secretory cavities of Japanese pepper leaves, but not in other parts of leaf tissues that do not include secretory cavities. We have obtained cDNAs for ZpTPS1 and ZpTPS2, which are responsible for biosynthesis of the sesquiterpenes β-caryophyllene and germacrene D, respectively, in Japanese pepper. In addition, we also identified a cDNA for the monoterpene synthase ZpTPS3. Expression of ZpTPS3 in Escherichia coli in addition to Agrobacterium-mediated transient ZpTPS3 expression in Nicotiana benthamiana demonstrated the catalytic activity of ZpTPS3 to form β-phellandrene as the major product. In situ hybridization in Japanese pepper leaf tissue revealed that ZpTPS3 transcript specifically accumulated in the epithelial cells surrounding secretory cavities. Expression of ZpTPS3 in epithelial cells was only detectable during early stages of cavity development, whereas the formation of volatile terpenes occurred at a constant rate throughout the expansion of secretory cavities. Our studies have improved the understanding of the currently uncharacterized processes controlling volatile terpene biosynthesis in Japanese pepper leaves.

In planta variation of volatile biosynthesis: an alternative biosynthetic route to the formation of the pathogen-induced volatile homoterpene DMNT via triterpene degradation in Arabidopsis roots.

Plant-derived volatile compounds such as terpenes exhibit substantial structural variation and serve multiple ecological functions. Despite their structural diversity, volatile terpenes are generally produced from a small number of core 5- to 20-carbon intermediates. Here, we present unexpected plasticity in volatile terpene biosynthesis by showing that irregular homo/norterpenes can arise from different biosynthetic routes in a tissue specific manner. While Arabidopsis thaliana and other angiosperms are known to produce the homoterpene (E)-4,8-dimethyl-1,3,7-nonatriene (DMNT) or its C16-analog (E,E)-4,8,12-trimethyl-1,3,7,11-tridecatetraene by the breakdown of sesquiterpene and diterpene tertiary alcohols in aboveground tissues, we demonstrate that Arabidopsis roots biosynthesize DMNT by the degradation of the C30 triterpene diol, arabidiol. The reaction is catalyzed by the Brassicaceae-specific cytochrome P450 monooxygenase CYP705A1 and is transiently induced in a jasmonate-dependent manner by infection with the root-rot pathogen Pythium irregulare. CYP705A1 clusters with the arabidiol synthase gene ABDS, and both genes are coexpressed constitutively in the root stele and meristematic tissue. We further provide in vitro and in vivo evidence for the role of the DMNT biosynthetic pathway in resistance against P. irregulare. Our results show biosynthetic plasticity in DMNT biosynthesis in land plants via the assembly of triterpene gene clusters and present biochemical and genetic evidence for volatile compound formation via triterpene degradation in plants.

Isolation and characterization of terpene synthases in cotton (Gossypium hirsutum)

Cotton plants accumulate gossypol and related sesquiterpene aldehydes, which function as phytoalexins against pathogens and feeding deterrents to herbivorous insects. However, to date little is known about the biosynthesis of volatile terpenes in this crop. Herein is reported that 5 monoterpenes and 11 sesquiterpenes from extracts of a glanded cotton cultivar, Gossypium hirsutum cv. CCRI12, were detected by gas chromatography-mass spectrometry (GC-MS). By EST data mining combined with Rapid Amplification of cDNA Ends (RACE), full-length cDNAs of three terpene synthases (TPSs), GhTPS1, GhTPS2 and GhTPS3 were isolated. By in vitro assays of the recombinant proteins, it was found that GhTPS1 and GhTPS2 are sesquiterpene synthases: the former converted farnesyl pyrophosphate (FPP) into β-caryophyllene and α-humulene in a ratio of 2:1, whereas the latter produced several sesquiterpenes with guaia-1(10),11-diene as the major product. By contrast, GhTPS3 is a monoterpene synthase, which produced α-pinene, β-pinene, β-phellandrene and trace amounts of other monoterpenes from geranyl pyrophosphate (GPP). The TPS activities were also supported by Virus Induced Gene Silencing (VIGS) in the cotton plant. GhTPS1 and GhTPS3 were highly expressed in the cotton plant overall, whereas GhTPS2 was expressed only in leaves. When stimulated by mechanical wounding, Verticillium dahliae (Vde) elicitor or methyl jasmonate (MeJA), production of terpenes and expression of the corresponding synthase genes were induced. These data demonstrate that the three genes account for the biosynthesis of volatile terpenes of cotton, at least of this Upland cotton.

Characterization of three novel isoprenyl diphosphate synthases from the terpenoid rich mango fruit

Mango (cv. Alphonso) is popular due to its highly attractive, terpenoid-rich flavor. Although Alphonso is clonally propagated, its fruit-flavor composition varies when plants are grown in different geo-climatic zones. Isoprenyl diphosphate synthases catalyze important branch-point reactions in terpenoid biosynthesis, providing precursors for common terpenoids such as volatile terpenes, sterols and carotenoids. Two geranyl diphosphate synthases and a farnesyl diphosphate synthase were isolated from Alphonso fruits, cloned for recombinant expression and found to produce the respective products. Although, one of the geranyl diphosphate synthases showed high sequence similarity to the geranylgeranyl diphosphate synthases, it did not exhibit geranylgeranyl diphosphate synthesizing activity. When modeled, this geranyl diphosphate synthase and farnesyl diphosphate synthase structures were found to be homologous with the reference structures, having all the catalytic side chains appropriately oriented. The optimum temperature for both the geranyl diphosphate synthases was 40 °C and that for farnesyl diphosphate synthase was 25 °C. This finding correlated well with the dominance of monoterpenes in comparison to sesquiterpenes in the fruits of Alphonso mango in which the mesocarp temperature is higher during ripening than development. The absence of activity of these enzymes with the divalent metal ion other than Mg(2+) indicated their adaptation to the Mg(2+) rich mesocarp. The typical expression pattern of these genes through the ripening stages of fruits from different cultivation localities depicting the highest transcript levels of these genes in the stage preceding the maximum terpene accumulation indicated the involvement of these genes in the biosynthesis of volatile terpenes.

The effects of exogenous methyl jasmonate on FaNES1 gene expression and the biosynthesis of volatile terpenes in strawberry (Fragaria × ananassa Duch.) fruit

SummaryThe volatile terpenoid profiles of cultivated strawberry (Fragaria × ananassa Duch.) fruit are dominated by the monoterpene, linalool, and the sesquiterpene, nerolidol. These terpenoids, generated by the product of the F. × ananassa nerolidol synthase1 (FaNES1) gene, are important for the aroma of strawberry fruit. FaNES1 gene transcript levels and emissions of linalool and nerolidol were measured at various developmental stages in two strawberry cultivars (‘Yan Xiang’ and ‘Shu Xiang’). Up-regulation of FaNES1 gene expression increased the emission of both terpenoids, depending on the degree of fruit ripeness. Terpenoid emissions were highest in red-mature (R) fruit. In addition, exogenous application of methyl jasmonate (MeJA) resulted in cultivar-dependent changes in the level of FaNES1 gene expression and terpenoid emissions. ‘Yan Xiang’ was sensitive to high concentrations of MeJA (i.e., 10.0 or 100.0 µM), while ‘Shu Xiang’ responded to low MeJA concentrations (1.0 µM). These results suggest that there might be a positive relationship between the emission of volatile aroma terpenoids and levels of FaNES1 gene transcripts. Furthermore, the effect of MeJA on increasing the emission of volatiles might be mediated by the induction of FaNES1 gene expression.

Four terpene synthases produce major compounds of the gypsy moth feeding-induced volatile blend of Populus trichocarpa

After herbivore damage, many plants increase their emission of volatile compounds, with terpenes usually comprising the major group of induced volatiles. Populus trichocarpa is the first woody species with a fully sequenced genome, enabling rapid molecular approaches towards characterization of volatile terpene biosynthesis in this and other poplar species. We identified and characterized four terpene synthases (PtTPS1–4) from P. trichocarpa which form major terpene compounds of the volatile blend induced by gypsy moth ( Lymantria dispar) feeding. The enzymes were heterologously expressed and assayed with potential prenyl diphosphate substrates. PtTPS1 and PtTPS2 accepted only farnesyl diphosphate and produced (−)-germacrene D and ( E, E)-α-farnesene as their major products, respectively. In contrast, PtTPS3 and PtTPS4 showed both mono- and sesquiterpene synthase activity. They produce the acyclic terpene alcohols linalool and nerolidol but exhibited opposite stereospecificity. qRT-PCR analysis revealed that the expression of the respective terpene synthase genes was induced after feeding of gypsy moth caterpillars. The TPS enzyme products may play important roles in indirect defense of poplar to herbivores and in mediating intra- and inter-plant signaling.

Insect-induced gene expression at the core of volatile terpene release in Medicago truncatula

The blends of induced volatiles released by higher plants in response to herbivory regularly contain terpenoids. The precursors of volatile terpenoids can be synthesized via two pathways, the mevalonate (MVA) and the methyl erythritol 4-phosphate (MEP) pathways localized in the cytosol and in plastids, respectively. Terpenes are important players in interactions between plants and herbivorous insects, by acting in both direct and indirect defenses. We recently characterized a gene encoding an (E)-β-ocimene synthase (MtEBOS) in the legume Medicago truncatula Gaertn. Compared to undamaged plants, caterpillar-damaged M. truncatula emitted (E)-β-ocimene at an elevated level and this increase is associated with high levels of expression of MtEBOS mRNA. Exogenous treatment with jasmonic acid also increases transcript accumulation of MtEBOS. These results indicate that transcript accumulation is used as a tightly regulated mechanism to control (E)-β-ocimene emission. The data, along with additional findings in other species, illustrate that like most plant families legumes regulate the final steps of volatile terpene biosynthesis at the level of transcript induction.

Medicago truncatula (E)-β-ocimene synthase is induced by insect herbivory with corresponding increases in emission of volatile ocimene

The blends of induced volatiles released by higher plants in response to herbivory regularly contain terpenoids. The precursors of volatile terpenoids can be synthesized via two pathways, the mevalonate (MVA) and the methyl erythritol 4-phosphate (MEP) pathways localized in the cytosol and in plastids, respectively. Terpenes are important players in interactions between plants and herbivorous insects, by acting in both direct and indirect defenses. We recently characterized a gene encoding an (E)-beta-ocimene synthase (MtEBOS) in the legume Medicago truncatula Gaertn. Compared to undamaged plants, caterpillar-damaged M. truncatula emitted (E)-beta-ocimene at an elevated level and this increase is associated with high levels of expression of MtEBOS mRNA. Exogenous treatment with jasmonic acid also increases transcript accumulation of MtEBOS. These results indicate that transcript accumulation is used as a tightly regulated mechanism to control (E)-beta-ocimene emission. The data, along with additional findings in other species, illustrate that like most plant families legumes regulate the final steps of volatile terpene biosynthesis at the level of transcript induction.

Induction of de Novo Volatile Terpene Biosynthesis via Cytosolic and Plastidial Pathways by Methyl Jasmonate in Foliage of Vitis vinifera L.

The terpene biosynthesis in leaves of Vitis vinifera L. cv. Morio Muskat was studied using methyl jasmonate to induce defensive responses in vivo. The experiments demonstrated the strong activation of the de novo biosynthesis of terpenoids via the octadecanoid-signaling cascade and release of the compounds to the gas phase. Feeding experiments with [5,5-(2)H(2)]-1-deoxy-d-xylulose and [5,5-(2)H(2)]mevalonic acid lactone allowed the investigation of the dynamic allocation of resources via the mevalonic acid and 1-deoxy-d-xylulose/2-C-methyl-d-erythritol 4-phosphate (DOXP/MEP) pathway under induced conditions and after treatment with the specific inhibitors mevastatin and fosmidomycin. The experiments reveal that monoterpenes are almost exclusively synthesized via the DOXP/MEP pathway, whereas sesquiterpenes are generated via both pathways at approximately equal rates. The biosynthesis of the homoterpene (E)-4,8-dimethyl-1,3,7-nonatriene was not affected by mevastatine or fosmidomycin.

Induction of volatile terpene biosynthesis and diurnal emission by methyl jasmonate in foliage of Norway spruce.

Terpenoids are characteristic constitutive and inducible defense chemicals of conifers. The biochemical regulation of terpene formation, accumulation, and release from conifer needles was studied in Norway spruce [Picea abies L. (Karst)] saplings using methyl jasmonate (MeJA) to induce defensive responses without inflicting physical damage to terpene storage structures. MeJA treatment caused a 2-fold increase in monoterpene and sesquiterpene accumulation in needles without changes in terpene composition, much less than the 10- and 40-fold increases in monoterpenes and diterpenes, respectively, observed in wood tissue after MeJA treatment (D. Martin, D. Tholl, J. Gershenzon, J. Bohlmann [2002] Plant Physiol 129: 1003-1018). At the same time, MeJA triggered a 5-fold increase in total terpene emission from foliage, with a shift in composition to a blend dominated by oxygenated monoterpenes (e.g. linalool) and sesquiterpenes [e.g. (E)-beta-farnesene] that also included methyl salicylate. The rate of linalool emission increased more than 100-fold and that of sesquiterpenes increased more than 30-fold. Emission of these compounds followed a pronounced diurnal rhythm with the maximum amount released during the light period. The major MeJA-induced volatile terpenes appear to be synthesized de novo after treatment, rather than being released from stored terpene pools, because they are almost completely absent from needle oleoresin and are the major products of terpene synthase activity measured after MeJA treatment. Based on precedents in other species, the induced emission of terpenes from Norway spruce foliage may have ecological and physiological significance.