Emissions Of Green Leaf Volatiles Research Articles

Impact of severe drought on biogenic volatile organic compounds emissions from Sphagnum mosses in boreal peatlands

Climate change and the associated increased frequency of extreme weather events are likely to alter the emissions of biogenic volatile organic compounds (BVOCs) from boreal peatlands. Hydrologically sensitive Sphagnum mosses are keystone species in boreal peatland ecosystems that are known to emit various BVOCs. However, it is not known how their emissions respond to seasonal droughts. In this study, we quantified the effect of severe drought, and subsequent recovery, on the BVOC emissions from Sphagnum mosses using mesocosms originating from wet open and naturally drier treed boreal fens and bogs. Here we report the emissions of 30 detected BVOCs, of which isoprene was the most abundant with an average flux rate of 5.6 μg m−2 h−1 (range 0–31.9 μg m−2 h−1). The experimental 43-day ecohydrological drought reduced total BVOC and isoprene emissions. In addition, in mesocosms originating from bogs, sesquiterpene emissions decreased with the drought, while the emissions of green leaf volatiles were induced. Sesquiterpene emissions remained low even six weeks after rewetting, indicating a long and limited recovery from the drought. Our results further imply that long-term exposure to deep water tables does not decrease sensitivity of Sphagnum to an extreme drought; we did not detect differences in the emission rates or drought responses between Sphagna originating from wet open and naturally drier treed habitats. Yet, the differences between fen and bog originating Sphagna indicate local variability in the BVOC quality changes following drought, potentially altering the climate feedback of boreal peatland BVOC emissions.

Ozone stress response of leaf BVOC emission and photosynthesis in mountain birch (Betula pubescens spp. czerepanovii) depends on leaf age.

Oxidative stress from ozone (O3) causes plants to alter their emission of biogenic volatile organic compounds (BVOC) and their photosynthetic rate. Stress reactions from O3 on birch trees can result in prohibited plant growth and lead to increased BVOC emission rates as well as changes in their compound blend to emit more monoterpenes (MT) and sesquiterpenes (SQT). BVOCs take part in atmospheric reactions such as enhancing the production of secondary organic aerosols (SOA). As the compound blend and emission rate change with O3 stress, this can influence the atmospheric conditions by affecting the production of SOA. Studying the stress responses of plants provides important information on how these reactions might change, which is vital to making better predictions of the future climate. In this study, measurements were taken to find out how the leaves of mature mountain birch trees (Betula pubescens ssp. czerepanovii) respond to different levels of elevated O3 exposure insitu depending on leaf age. We found that leaves from both early and late summers responded with induced SQT emission after exposure to 120 ppb O3. Early leaves were, however, more sensitive to increased O3 concentrations, with enhanced emission of green leaf volatiles (GLV) and tendencies of both induced leaf senescence as well as poor recovery in the photosynthetic rate between exposures. Late leaves had more stable photosynthetic rates throughout the experiment and responded less to exposure at different O3 levels.

The beneficial effects of the tobacco hydroperoxide lyase pathway in whitefly host adaptation

Plants respond to herbivore attack by emitting complex mixtures of volatile compounds to repel herbivores or attract predators and parasitoids. In this study, however, we revealed that tobacco green leaf volatiles (GLVs) were beneficial compounds for whitefly adaptation to host plants. Our study indicates that GLVs are closely associated with plant susceptibility to whiteflies. Whitefly infestation elevated the transcript of the hydroperoxide lyase (HPL) gene, one gene responsible for catalyzing the synthesis of C6 or C9 aldehydes from fatty acid hydroperoxides. Overexpression of HPL increased the emission of GLVs (hexanal, 1- hexanal, trans-2- hexanal and cis-3- hexanal) and resulted in improved performance for whiteflies. Exogenous application of GLVs promoted whitefly survival and fecundity and increased the plant's attraction to whiteflies. Thus, our study provides new insights into the role of the HPL pathway and GLVs in influencing herbivore adaptation to host plants.

Seasonal and Elevational Variability in Dwarf Birch VOC Emissions in Greenlandic Tundra

AbstractRising temperatures amplify biogenic volatile organic compound (VOC) emissions from Arctic vegetation, causing feedbacks to the climate system. Changes in climate also alter plant physiology and vegetation composition, all of which can influence VOC emissions. Moreover, leaf development and biotic stresses cause highly variable emissions during the growing season. Therefore, linking VOC emissions with plant traits and tracking responses to climate change might provide better understanding of VOC emission regulation under future conditions. We measured VOC emissions and other plant traits in dwarf birch (Betula glandulosa) at two elevations in Narsarsuaq, South Greenland. The measurements were performed in warming experiments that have run since 2016. We collected VOCs using the branch enclosure method from early June until late July 2019 (n = 200). Emissions of green leaf volatiles (GLVs), oxygenated monoterpenes (oMTs), and homoterpenes followed a seasonal trend. VOC emission rates and the diversity of the VOC blend decreased at the end of the measurement period. Differences in VOC emission rates between elevations were pronounced early in the season. Majority of the traits did not explain the variation in VOC emissions. We show strong seasonal variability in VOC emissions within the growing season, which is likely driven by leaf phenology. While the diversity of VOCs was greater at the milder low‐elevation site, VOC emission rates were higher or similar at the harsher high‐elevation site, showing stronger VOC emission potentials than previously assumed. Seasonal variations in the emissions of VOCs are crucial for accurate predictions of current and future VOC emissions from arctic ecosystems.

Dynamic distress calls: volatile info chemicals induce and regulate defense responses during herbivory.

Plants are continuously threatened by a plethora of biotic stresses caused by microbes, pathogens, and pests, which often act as the major constraint in crop productivity. To overcome such attacks, plants have evolved with an array of constitutive and induced defense mechanisms- morphological, biochemical, and molecular. Volatile organic compounds (VOCs) are a class of specialized metabolites that are naturally emitted by plants and play an important role in plant communication and signaling. During herbivory and mechanical damage, plants also emit an exclusive blend of volatiles often referred to as herbivore-induced plant volatiles (HIPVs). The composition of this unique aroma bouquet is dependent upon the plant species, developmental stage, environment, and herbivore species. HIPVs emitted from infested and non-infested plant parts can prime plant defense responses by various mechanisms such as redox, systemic and jasmonate signaling, activation of mitogen-activated protein (MAP) kinases, and transcription factors; mediate histone modifications; and can also modulate the interactions with natural enemies via direct and indirect mechanisms. These specific volatile cues mediate allelopathic interactions leading to altered transcription of defense-related genes, viz., proteinase inhibitors, amylase inhibitors in neighboring plants, and enhanced levels of defense-related secondary metabolites like terpenoids and phenolic compounds. These factors act as deterrents to feeding insects, attract parasitoids, and provoke behavioral changes in plants and their neighboring species. This review presents an overview of the plasticity identified in HIPVs and their role as regulators of plant defense in Solanaceous plants. The selective emission of green leaf volatiles (GLVs) including hexanal and its derivatives, terpenes, methyl salicylate, and methyl jasmonate (MeJa) inducing direct and indirect defense responses during an attack from phloem-sucking and leaf-chewing pests is discussed. Furthermore, we also focus on the recent developments in the field of metabolic engineering focused on modulation of the volatile bouquet to improve plant defenses.

Green Leaf Volatile Function in Both the Natural Defense System of Rumex confertus and Associated Insects’ Behavior

Rumex confertus is a rhizomatous, invasive, and difficult to control plant. Nevertheless, for sustainable agriculture, studies to biologically control R. confertus continue. Towards this, considerable attention has been devoted to investigating the emission of a wide array of volatile organic compounds (VOCs) from herbivore-damaged plants, which are known to induce protection measures in neighboring, undamaged plants. The goals of this study are to (1) determine if the profile of green leaf volatiles (GLVs), which are organic compounds naturally produced by undamaged plants, is similar to that provoked by the chemical stimulants Z-jasmone (ZJA) and dihydrojasmone (DJA), (2) establish if the Apion miniatum beetle’s reproductive choices are influenced by their sex and mating status, and (3) assess if chemically stimulated GLV emissions can be used as signals to attract pests to R. confertus for biological control purposes. Synthetic forms of naturally produced Z-jasmone (ZJA) and dihydrojasmone (DJA), which can act as either an attractant or a repellent of various species of insects, were used to treat R. confertus. In olfactory analysis, the behavioral responses of individual insects to mated and unmated insects and to two GLV blends were evaluated. It was found that unmated male insects were fairly equally divided between going for food (Y-tube olfactometer arm with a GLV blend) and opportunities for reproduction (Y-tube olfactometer arm with unmated females). However, an increase in the GLV blend concentration attracted the insects. Meanwhile, unmated females were definitely indifferent to food and, independent of the GLV blend dose, were more interested in reproduction. Mated males, even with weak feed stimuli, increased their reproduction activity, in opposition to mated females. We provide evidence that synthetic GLV blends can be used to attract predators, making their use an effective means to biologically control R. confertus. The idea of applying synthetic GLVs as a biological control is based on the insects’ mutual relationships, which work as follows: (i) mated males fully invade the weed, (ii) reproduction-driven females follow the mated males to R. confertus, and (iii) the unmated males follow the females with the purpose to reproduce. Therefore, all insect groups (mated and unmated males and females) can be induced to invade the weed. Upon feeding, the insects will damage the generative organs of R. confertus. We propose that the use of chemical stimulants to increase GLV emissions can be effectively used for weed (here R. confertus) control by attracting a plant pest (here A. miniatum).

Volatile responses of dwarf birch to mimicked insect herbivory and experimental warming at two elevations in Greenlandic tundra.

Plants release a complex blend of volatile organic compounds (VOCs) in response to stressors. VOC emissions vary between contrasting environments and increase with insect herbivory and rising temperatures. However, the joint effects of herbivory and warming on plant VOC emissions are understudied, particularly in high latitudes, which are warming fast and facing increasing herbivore pressure. We assessed the individual and combined effects of chemically mimicked insect herbivory, warming, and elevation on dwarf birch (Betula glandulosa) VOC emissions in high-latitude tundra ecosystems in Narsarsuaq, South Greenland. We hypothesized that VOC emissions and compositions would respond synergistically to warming and herbivory, with the magnitude differing between elevations. Warming increased emissions of green leaf volatiles (GLVs) and isoprene. Herbivory increased the homoterpene, (E)-4,8-dimethyl-1,3,7-nonatriene, emissions, and the response was stronger at high elevation. Warming and herbivory had synergistic effects on GLV emissions. Dwarf birch emitted VOCs at similar rates at both elevations, but the VOC blends differed between elevations. Several herbivory-associated VOC groups did not respond to herbivory. Harsher abiotic conditions at high elevations might not limit VOC emissions from dwarf birch, and high-elevation plants might be better at herbivory defense than assumed. The complexity of VOC responses to experimental warming, elevation, and herbivory are challenging our understanding and predictions of future VOC emissions from dwarf birch-dominated ecosystems.

Acute methyl jasmonate exposure results in major bursts of stress volatiles, but in surprisingly low impact on specialized volatile emissions in the fragrant grass Cymbopogon flexuosus

Methyl jasmonate (MeJA) is an airborne hormonal elicitor that induces a fast rise of emissions of characteristic stress marker compounds methanol and green leaf volatiles (GLV), and a longer-term release of volatile terpenoids, but there is limited information of how terpene emissions respond to MeJA in terpene-storing species. East-Indian lemongrass (Cymbopogon flexuosus), an aromatic herb with a large terpenoid storage pool in idioblasts, was used to investigate the short- (0–1 h) and long-term (1–16 h) responses of leaf net assimilation rate (A), stomatal conductance (Gs) and volatile emissions to MeJA concentrations ranging from moderate to lethal. Both A and Gs were increasingly inhibited with increasing MeJA concentration in both short and long term. MeJA exposure resulted in a rapid elicitation, within 1 h after exposure, of methanol and GLV emissions. Subsequently, a secondary rise of GLV emissions was observed, peaking at 2 h after MeJA exposure for the highest and at 8 h for the lowest application concentration. The total amount and maximum emission rate of methanol and the first and second GLV emission bursts were positively correlated with MeJA concentration. Unexpectedly, no de novo elicitation of terpene emissions was observed through the experiment. Although high MeJA application concentrations led to visible lesions and desiccation in extensive leaf regions, this did not result in breakage of terpene-storing idioblasts. The study highlights an overall insensitivity of lemongrass to MeJA and indicates that differently from mechanical wounding, MeJA-driven cellular death does not break terpene-storing cells. Further studies are needed to characterize the sensitivity of induced defense responses in species with strongly developed constitutive defenses.

Herbivorous Caterpillars and the Green Leaf Volatile (GLV) Quandary.

Several herbivorous caterpillars contain effectors in their oral secretions that alter the emission of green leaf volatiles (GLVs) produced by the plants upon which the caterpillars are feeding. These effectors include an isomerase, a fatty acid dehydratase (FHD), and a heat-stable hexenal trapping (HALT) molecule. GLVs serve as signaling compounds in plant-insect interactions and inter-and intra-plant communication. However, it is not known whether these GLV-altering effectors are common among herbivorous caterpillars, or the evolutionary context of these effectors in relation to GLV emission by host plants in response to feeding damage. Here, we examined the distribution and activity of the isomerase, FHD, and HALT effectors across 10 species spanning 7 lepidopteran families. Six of the 10 species possessed all three effectors in their oral secretions. Activity from the HALT and FHD effectors was observed in all examined caterpillar species, while activity from the isomerase effector varied in some species and was absent in others. There was no discernable pattern in effector activity based on evolutionary divergence, since individual species within a family did not possess similar mechanisms to alter GLV emission. These data, demonstrating the GLV-altering effectors acting at different steps in the GLV biosynthetic pathway and present in the examined caterpillar species at different combinations with different activities, highlight the importance of these effectors in changing the emission of these compounds during caterpillar herbivory. Understanding the prevalence and roles of GLV-altering effectors and GLV emission itself will open new research areas in the dynamics of plant-insect interactions.

Light dominates the diurnal emissions of herbivore-induced volatiles in wild tobacco

BackgroundTiming is everything when it comes to the fitness outcome of a plant’s ecological interactions, and accurate timing is particularly relevant for interactions with herbivores or mutualists that are based on ephemeral emissions of volatile organic compounds. Previous studies of the wild tobacco N. attenuata have found associations between the diurnal timing of volatile emissions, and daytime predation of herbivores by their natural enemies.ResultsHere, we investigated the role of light in regulating two biosynthetic groups of volatiles, terpenoids and green leaf volatiles (GLVs), which dominate the herbivore-induced bouquet of N. attenuata. Light deprivation strongly suppressed terpenoid emissions while enhancing GLV emissions, albeit with a time lag. Silencing the expression of photoreceptor genes did not alter terpenoid emission rhythms, but silencing expression of the phytochrome gene, NaPhyB1, disordered the emission of the GLV (Z)-3-hexenyl acetate. External abscisic acid (ABA) treatments increased stomatal resistance, but did not truncate the emission of terpenoid volatiles (recovered in the headspace). However, ABA treatment enhanced GLV emissions and leaf internal pools (recovered from tissue), and reduced internal linalool pools. In contrast to the pattern of diurnal terpenoid emissions and nocturnal GLV emissions, transcripts of herbivore-induced plant volatile (HIPV) biosynthetic genes peaked during the day. The promotor regions of these genes were populated with various cis-acting regulatory elements involved in light-, stress-, phytohormone- and circadian regulation.ConclusionsThis research provides insights into the complexity of the mechanisms involved in the regulation of HIPV bouquets, a mechanistic complexity which rivals the functional complexity of HIPVs, which includes repelling herbivores, calling for body guards, and attracting pollinators.

Sesquiterpenes and oxygenated sesquiterpenes dominate the VOC (C<sub>5</sub>–C<sub>20</sub>) emissions of downy birches

Abstract. Biogenic volatile organic compounds (BVOCs) emitted by the forests are known to have strong impacts in the atmosphere. However, lots of missing reactivity is found, especially in the forest air. Therefore better characterization of sources and identification/quantification of unknown reactive compounds is needed. While isoprene and monoterpene (MT) emissions of boreal needle trees have been studied quite intensively, there is much less knowledge on the emissions of boreal deciduous trees and emissions of larger terpenes and oxygenated volatile organic compounds (OVOCs). Here we quantified the downy birch (Betula pubescens) leaf emissions of terpenes, oxygenated terpenes and green leaf volatiles (GLVs) at the SMEAR II boreal forest site using in situ gas chromatographs with mass spectrometers. Sesquiterpenes (SQTs) and oxygenated sesquiterpenes (OSQTs) were the main emitted compounds. Mean emission rates of SQTs and OSQTs were significantly higher in the early growing season (510 and 650 ng gdw-1 h−1, respectively) compared to in the main (40 and 130 ng gdw-1 h−1, respectively) and late (14 and 46 ng gdw-1 h−1, respectively) periods, indicating that early leaf growth is a strong source of these compounds. The emissions had a very clear diurnal variation with afternoon maxima being on average 4 to 8 times higher than seasonal means for SQTs and OSQTs, respectively. β-Caryophyllene and β-farnesene were the main SQTs emitted. The main emitted OSQTs were tentatively identified as 14-hydroxy-β-caryophyllene acetate (M=262 g mol−1) and 6-hydroxy-β-caryophyllene (M=220 g mol−1). Over the whole growing season, the total MT emissions were only 24 % and 17 % of the total SQT and OSQT emissions, respectively. A stressed tree growing in a pot was also studied, and high emissions of α-farnesene and an unidentified SQT were detected together with high emissions of GLVs. Due to the relatively low volatility and the high reactivity of SQTs and OSQTs, downy birch emissions are expected to have strong impacts on atmospheric chemistry, especially on secondary organic aerosol (SOA) production.

Eucalyptus species maintain secondary metabolite production under water stress conditions at the expense of growth

AbstractPlants have evolved a vast array of secondary metabolites that help defend them against natural enemy attack. The production of these secondary metabolites is influenced by environmental factors, such as temperature, rainfall (soil water availability) and atmospheric [CO2], which may be altered under climate change. This study aimed to test how two of these environmental factors, namely soil water availability and [CO2], will affect secondary metabolite and biomass production in Eucalypts. To investigate this, we measured secondary metabolite and biomass production of three Eucalypt species (Eucalyptus grandis, E. moluccana, E. saligna) grown under ambient and elevated [CO2] (400 and 600 ppm, respectively) and water‐stressed and well‐watered conditions, in a fully factorial glasshouse experiment. The secondary metabolites measured were foliar phenolic concentration as well as terpene and green leaf volatile emissions. We found that water stress did not alter secondary metabolite production. Unsurprisingly, the water‐stressed plants maximised their water uptake potential by reducing shoot biomass but maintaining root biomass. The detrimental effects of water stress on shoot biomass production were not reduced under elevated [CO2] compared to ambient [CO2], despite all species experiencing significant reductions in stomatal conductance. Further, [CO2] as a single factor did not have a significant effect on species’ net carbon gain, with an increase in green leaf volatile emissions under elevated [CO2] being mitigated by a reduction in shoot biomass production. These results suggest that reductions in carbon gains due to water stress will affect the growth of Eucalypts rather than their chemical defence capabilities.

Influence of Brevibacterium linens RS16 on foliage photosynthetic and volatile emission characteristics upon heat stress in Eucalyptus grandis

Heat stress induces secondary metabolic changes in plants, channeling photosynthetic carbon and energy, away from primary metabolic processes, including, growth. Use of ACC (1-aminocyclopropane-1-carboxylate) deaminase containing plant growth promoting bacteria (PGPB) in conferring heat resistance in plants and the role of PGPB, in altering net carbon assimilation, constitutive and stress volatile emissions has not been studied yet. We exposed leaves of Eucalyptus grandis inoculated and non-inoculated with PGPB Brevibacterium linens RS16 to two levels of heat stress (37 °C and 41 °C for 5 min) and quantified temporal changes in foliage photosynthetic characteristics and volatile emission rates at 0.5 h, day 1 and day 5 after the stress application. Heat stress resulted in immediate reductions in dark-adapted photosystem II (PSII) quantum yield (Fv/Fm), net assimilation rate (A), stomatal conductance to water vapor (gs), and enhancement of stress volatile emissions, including enhanced emissions of green leaf volatiles (GLV), mono- and sesquiterpenes, light weight oxygenated volatile organic compounds (LOC), geranyl-geranyl diphosphate pathway volatiles (GGDP), saturated aldehydes, and benzenoids, with partial recovery by day 5. Changes in stress-induced volatiles were always less in leaves inoculated with B. linens RS16. However, net assimilation rate was enhanced by bacterial inoculation only in the 37 °C treatment and overall reduction of isoprene emissions was observed in bacterially-treated leaves. Principal component analysis (PCA), correlation analysis and partial least squares discriminant analysis (PLS-DA) indicated that different stress applications influenced specific volatile organic compounds. In addition, changes in the expression analysis of heat shock protein 70 gene (DnaK) gene in B. linens RS16 upon exposure to higher temperatures further indicated that B. linens RS16 has developed its own heat resistance mechanism to survive under higher temperature regimes. Taken together, this study demonstrates that foliar application of ACC deaminase containing PGPB can ameliorate heat stress effects in realistic biological settings.

An unbiased approach elucidates variation in (S)-(+)-linalool, a context-specific mediator of a tri-trophic interaction in wild tobacco

Plant volatile organic compounds (VOCs) mediate many interactions, and the function of common VOCs is especially likely to depend on ecological context. We used a genetic mapping population of wild tobacco, Nicotiana attenuata, originating from a cross of 2 natural accessions from Arizona and Utah, separated by the Grand Canyon, to dissect genetic variation controlling VOCs. Herbivory-induced leaf terpenoid emissions varied substantially, while green leaf volatile emissions were similar. In a field experiment, only emissions of linalool, a common VOC, correlated significantly with predation of the herbivore Manduca sexta by native predators. Using quantitative trait locus mapping and genome mining, we identified an (S)-(+)-linalool synthase (NaLIS). Genome resequencing, gene cloning, and activity assays revealed that the presence/absence of a 766-bp sequence in NaLIS underlies the variation of linalool emissions in 26 natural accessions. We manipulated linalool emissions and composition by ectopically expressing linalool synthases for both enantiomers, (S)-(+)- and (R)-(-)-linalool, reported to oppositely affect M. sexta oviposition, in the Arizona and Utah accessions. We used these lines to test ovipositing moths in increasingly complex environments. The enantiomers had opposite effects on oviposition preference, but the magnitude of the effect depended strongly both on plant genetic background, and complexity of the bioassay environment. Our study reveals that the emission of linalool, a common VOC, differs by orders-of-magnitude among geographically interspersed conspecific plants due to allelic variation in a linalool synthase, and that the response of a specialist herbivore to linalool depends on enantiomer, plant genotype, and environmental complexity.

Specificity of Herbivore Defense Responses in a Woody Plant, Black Poplar (Populus nigra)

The specificity of woody plant defense responses to different attacking herbivores is poorly known. We investigated the responses of black poplar (Populus nigra) to leaf feeding by three lepidopteran species (Lymantria dispar, Laothoe populi and Amata mogadorensis) and two leaf beetle species (Phratora vulgatissima and Chrysomela populi). Of the direct defenses monitored, increases in trypsin protease inhibitor activity and the salicinoid salicin were triggered by herbivore damage, but this was not herbivore-specific. Moreover, the majority of leaf salicinoid content was present constitutively and not induced by herbivory. On the other hand, volatile emission profiles did vary among herbivore species, especially between coleopterans and lepidopterans. Monoterpenes and sesquiterpenes were induced in damaged and adjacent undamaged leaves, while the emission of green leaf volatiles, aromatic and nitrogen-containing compounds (known to attract herbivore enemies) was restricted to damaged leaves. In conclusion, indirect defenses appear to show more specific responses to attacking herbivores than direct defenses in this woody plant.

Herbivorous Caterpillars Can Utilize Three Mechanisms to Alter Green Leaf Volatile Emission.

Green plants emit green leaf volatiles (GLVs) as a general damage response. These compounds act as signals for the emitter plant, neighboring plants, and even for insects in the ecosystem. However, when oral secretions from certain caterpillars are applied to wounded leaves, GLV emissions are significantly decreased or modified. We examined four caterpillar species representing two lepidopteran families for their capacity to decrease GLV emissions from Zea mays leaf tissue. We also investigated the source of the GLV modifying components in the alimentary tract of the various caterpillars. In Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae), Spodoptera frugiperda (Smith) (Lepidoptera: Noctuidae), Trichoplusia ni (Hübner) (Lepidoptera: Noctuidae), and Manduca sexta (Linnaeus) (Lepidoptera: Sphingidae), we found three distinct mechanisms to modify GLV emission: a heat-stable compound in the gut, a heat-labile enzyme in salivary gland homogenate (previously described in Bombyx mori (Linnaeus) (Lepidoptera: Bombycidae), and an isomerase in the salivary gland homogenate, which catalyzes the conversion of (Z)-3-hexenal to (E)-2-hexenal (previously described in M. sexta). These mechanisms employed by caterpillars to suppress or modify GLV emission suggest a counteraction against the induced indirect volatile defenses of a plant and provides further insights into the ecological functions of GLVs.

Characterization of Plant Volatiles Reveals Distinct Metabolic Profiles and Pathways among 12 Brassicaceae Vegetables.

Plants emit characteristic organic volatile compounds (VOCs) with diverse biological/ecological functions. However, the links between plant species/varieties and their phytochemical emission profiles remain elusive. Here, we developed a direct headspace solid-phase microextraction (HS-SPME) technique and combined with non-targeted gas chromatography‒high-resolution mass spectrometry (GC-HRMS) platform to investigate the VOCs profiles of 12 common Brassicaceae vegetables (watercress, rocket, Brussels sprouts, broccoli, kai lan, choy sum, pak choi, cabbage, Chinese cabbage, cauliflower, radish and cherry radish). The direct HS-SPME sampling approach enabled reproducible capture of the rapid-emitting VOCs upon plant tissue disruption. The results revealed extensive variation in VOCs profiles among the 12 Brassicaceae vegetables. Furthermore, principal component analysis (PCA) showed that the VOC profiles could clearly distinguish the 12 Brassicaceae vegetables, and that these profiles well reflected the classical morphological classification. After multivariate statistical analysis, 44 VOCs with significant differences among the Brassicaceae vegetables were identified. Pathway analysis showed that three secondary metabolism pathways, including the fatty acid pathway, methylerythritol phosphate (MEP) pathway and glucosinolate (GLS) pathway, behave distinctively in these vegetables. These three pathways are responsible for the generation and emission of green leaf volatiles (GLVs), terpenes and isothiocyanates (ITCs), respectively. Correlation analysis further showed that volatile metabolites formed via the common pathway had significantly positive correlations, whereas metabolites from different pathways had either non-significant or significantly negative correlations. Genetic influences on these metabolites across various vegetable types were also evaluated. These findings extend our phytochemical knowledge of the 12 edible Brassicaceae vegetables and provide useful information on their secondary metabolism.

Herbivory elicits changes in green leaf volatile production via jasmonate signaling and the circadian clock.

The timing of plant volatile emissions is important for a robust indirect defense response. Green leaf volatiles (GLVs) are emitted by plants upon damage but can be suppressed by herbivore-associated elicitors, and the abundance and composition of GLVs vary depending on the timing of herbivore attack. We show that the GLV biosynthetic enzyme HYDROPEROXIDE LYASE (HPL) is transcriptionally regulated by the circadian clock in Nicotiana attenuata. In accordance with transcript abundance of NaHPL, GLV aldehyde pools in intact leaves peaked at night and at subjective night under diurnal and continuous light conditions, respectively. Moreover, although the basal abundance of NaHPL transcripts is upregulated by jasmonate (JA) signaling, JA does not regulate the reduction of NaHPL transcript abundance in damaged leaves by simulated herbivore treatment. Unexpectedly, the plant circadian clock was strongly altered when Manduca sexta larvae fed on N.attenuata, and this was also independent of JA signaling. Lastly, the temporal dynamics of NaHPL transcripts and total GLV emissions were strongly altered by M.sexta larval feeding. Our data suggest that the temporal dynamics of emitted GLV blends result from a combination of damage, JA signaling, herbivore-associated elicitors, and the plant circadian clock.

Silkworms suppress the release of green leaf volatiles by mulberry leaves with an enzyme from their spinnerets

In response to herbivory, plants emit a blend of volatile organic compounds that includes green leaf volatiles (GLVs) and terpenoids. These volatiles are known to attract natural enemies of herbivores and are therefore considered to function as an indirect defense. Selection should favor herbivores that are able to suppress these volatile emissions, and thereby make themselves less conspicuous to natural enemies. We tested this possibility for silkworms, which were observed to leave secretions from their spinnerets while feeding on mulberry leaves. When we ablated the spinnerets of silkworms, no secretions were observed. Leaves infested by intact silkworms released smaller amounts of GLVs than leaves infested by ablated silkworms, indicating that the spinneret secretion suppressed GLV production. This difference in GLV emissions was also reflected in the behavioral response of Zenillia dolosa (Tachinidae), a parasitoid fly of silkworms. The flies laid fewer eggs when exposed to the volatiles from intact silkworm-infested leaves than when exposed to the volatiles from ablated silkworm-infested leaves. We identified a novel enzyme in the secretion from the spinneret that is responsible for the GLV suppression. The enzyme converted 13(S)-hydroperoxy-(9Z,11E,15Z)-octadecatrienoic acid, an intermediate in the biosynthetic pathway of GLVs, into its keto-derivative in a stereospecific manner. Taken together, this study shows that silkworms are able to feed on mulberry in a stealthy manner by suppressing GLV production with an enzyme in secretions of their spinnerets, which might be a countermeasure against induced indirect defense by mulberry plants.

Green leaf volatile production by plants: a meta‐analysis

666 I. Introduction 667 II. Biosynthesis 667 III. Meta-analysis 669 IV. The type of stress influences the total amount of GLVs released 669 V. Herbivores can modulate the wound-induced release of GLVs 669 VI. Fungal infection greatly induces GLV production 672 VII. Monocots and eudicots respond differentially to different types of stress 673 VIII. The type of stress does not influence the proportion of GLVs per chemical class 673 IX. The type of stress does influence the isomeric ratio within each chemical class 674 X. GLVs: from signal perception to signal transduction 676 XI. GLVs influence the C/N metabolism 677 XII. Interaction with plant hormones 678 XIII. General conclusions and unanswered questions 678 Acknowledgements 679 References 679 SUMMARY: Plants respond to stress by releasing biogenic volatile organic compounds (BVOCs). Green leaf volatiles (GLVs), which are abundantly produced across the plant kingdom, comprise an important group within the BVOCs. They can repel or attract herbivores and their natural enemies; and they can induce plant defences or prime plants for enhanced defence against herbivores and pathogens and can have direct toxic effects on bacteria and fungi. Unlike other volatiles, GLVs are released almost instantly upon mechanical damage and (a)biotic stress and could thus function as an immediate and informative signal for many organisms in the plant's environment. We used a meta-analysis approach in which data from the literature on GLV production during biotic stress responses were compiled and interpreted. We identified that different types of attackers and feeding styles add a degree of complexity to the amount of emitted GLVs, compared with wounding alone. This meta-analysis illustrates that there is less variation in the GLV profile than we presumed, that pathogens induce more GLVs than insects and wounding, and that there are clear differences in GLV emission between monocots and dicots. Besides the meta-analysis, this review provides an update on recent insights into the perception and signalling of GLVs in plants.