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Herbivore-induced volatile production by Arabidopsis thaliana leads to attraction of the parasitoid Cotesia rubecula: chemical, behavioral, and gene-expression analysis.

Many plant species defend themselves against herbivorous insects indirectly by producing volatiles in response to herbivory. These volatiles attract carnivorous enemies of the herbivores. Research on the model plant Arabidopsis thaliana (L.) Heynh. has contributed considerably to the unraveling of signal transduction pathways involved in direct plant defense mechanisms against pathogens. Here, we demonstrate that Arabidopsis is also a good candidate for studying signal transduction pathways involved in indirect defense mechanisms by showing that: (1) Adult females of Cotesia rubecula, a specialist parasitic wasp of Pieris rapae caterpillars, are attracted to P. rapae-infested Arabidopsis plants. (2) Arabidopsis infested by P. rapae emits volatiles from several major biosynthetic pathways, including terpenoids and green leaf volatiles. The blends from herbivore-infested and artificially damaged plants are similar. However, differences can be found with respect to a few components of the blend, such as two nitriles and the monoterpene myrcene, that were produced exclusively by caterpillar-infested plants, and methyl salicylate, that was produced in larger amounts by caterpillar-infested plants. (3) Genes from major biosynthetic pathways involved in volatile production are induced by caterpillar feeding. These include AtTPS10, encoding a terpene synthase involved in myrcene production, AtPAL1, encoding phenylalanine ammonia-lyase involved in methyl salicylate production, and AtLOX2 and AtHPL, encoding lipoxygenase and hydroperoxide lyase, respectively, both involved in the production of green leaf volatiles. AtAOS, encoding allene oxide synthase, involved in the production of jasmonic acid, also was induced by herbivory.

Adaptation, Physiological↗

Chemical analysis of volatiles emitted by Pinus svlvestris after induction by insect oviposition.

Gas chromatography-mass spectrometry analyses of the headspace volatiles of Scots pine (Pinus sylvestris) induced by egg deposition of the sawfly Diprion pini were conducted. The odor blend of systemically oviposition-induced pine twigs. attractive for the eulophid egg parasitoid Chrysonotomyia ruforum, was compared to volatiles released by damaged pine twigs (control) that are not attractive for the parasitoid. The mechanical damage inflicted to the control twigs mimicked the damage by a sawfly female prior to egg deposition. The odor blend released by oviposition-induced pine twigs consisted of numerous mono- and sesquiterpenes, which all were also present in the headspace of the artificially damaged control twigs. A quantitative comparison of the volatiles from oviposition-induced twigs and controls revealed that only the amounts of (E)-beta-farnesene were significantly higher in the volatile blend of the oviposition-induced twigs. Volatiles from pine twigs treated with jasmonic acid (JA) also attract the egg parasitoid. No qualitative differences were detected when comparing the composition of the headspace of JA-treated pine twigs with the volatile blend of untreated control twigs. JA-treated pine twigs released significantly higher amounts of (E )-beta-farnesene. However, the JA treatment induced a significant increase of the amount of further terpenoid components. The release of terpenoids by pine after wounding, egg deposition, and JA treatment is discussed with special respect to (E)-beta-farnesene.

Adaptation, Physiological↗

Identification of volatile synomones, induced by Nezara viridula feeding and oviposition on bean spp., that attract the egg parasitoid Trissolcus basalis.

Bean plants (Vicia faba L. and Phaseolus vulgaris L.) damaged by feeding activity of Nezara viridula (L.) (Heteroptera: Pentatomidae), and onto which an egg mass had been laid, produced volatiles that attracted the egg parasitoid Trissolcus basalis (Wollaston) (Hymenoptera: Scelionidae). Extracts of volatiles of broad bean and French bean plants induced by adults of N. viridula as a result of their feeding activity, oviposition activity, and feeding and oviposition activity combined were analyzed by gas chromatography-mass spectrometry (GC-MS), and tested in Y-tube olfactometer bioassays as attractants for T. basalis females. In extracts from undamaged leguminous plants, green-leaf volatiles were absent or scarcely detected, and monoterpenes and sesquiterpenes were present at trace levels. No significant differences were detected in the profiles of volatiles of undamaged plants, and undamaged plants on which bugs were allowed only to lay eggs. In contrast, feeding and oviposition by adults of N. viridula induced in both leguminous plants a significant increase in terpenoids such as linalool, (E)-beta-caryophyllene, (E,E)-4,8,12-trimethyl-1,3,7,11-tridecatetraene, and (3E)-4,8-dimethyl-1,3,7-nonatriene, which was induced only in French bean plants. Quantitative comparisons revealed increased levels of (E)-beta-caryophyllene in extracts from feeding-damaged plants with N. viridula egg masses compared to feeding-damaged plants without egg masses. In Y-tube olfactometer bioassays, T. basalis females were attracted by extracts of both leguminous plants only when N. viridula adults were allowed to feed and oviposit upon them. Fractionation of extracts of volatiles from broad bean plants with N. viridula feeding damage and egg masses yielded two fractions. but only the fraction containing (E)-beta-caryophyllene was attractive to the egg parasitoid. These findings indicate that N. viridula feeding and oviposition induce leguminous plants to produce blends of volatiles that are characterized by increased amounts of (E)-beta-caryophyllene, and these blends attract female T. basalis. The role of (E)-beta-caryophyllene as a potential synomone for T. basalis is discussed.

Animals↗

Identification of volatiles that are used in discrimination between plants infested with prey or nonprey herbivores by a predatory mite.

Carnivorous arthropods can use herbivore-induced plant volatiles to locate their herbivorous prey. In the field, carnivores are confronted with information from plants infested with herbivores that may differ in their suitability as prey. Discrimination by the predatory mite Phytoseiulus persimilis between volatiles from lima bean plants infested with the prey herbivore Tetranychus urticae, or plants infested with the nonprey caterpillar Spodoptera exigua, depends on spider mite density. In this article, we analyzed the chemical composition of the volatile blends from T. urticae-infested lima bean plants at different densities of spider mites, and from S. exigua-infested plants. Based on the behavioral preferences of P. persimilis and the volatile profiles, we selected compounds that potentially enable the mite to discriminate between T. urticae-induced and S. exigua-induced volatiles. Subsequently, we demonstrated in Y-tube olfactometer assays that the relatively large amounts of methyl salicylate and (3E, 7E)-4,8,12-trimethyl-1,3,7,11-tridecatetraene emitted by T. urticae-infested bean plants compared to S. exigua-infested plants enable the predators to discriminate. Our data show that specific compounds from complex herbivore-induced volatile blends can play an important role in the selective foraging behavior of natural enemies of herbivorous arthropods.

Alkenes↗

Effects of volatile and nonvolatile chemical signals on male sex behaviors mediated by the main and accessory olfactory systems.

Estrous hamster vaginal discharge (HVD) contains both volatile and nonvolatile chemical signals which collectively elicit both male attraction to females and male mating behavior. These two aspects of normal sexual behavior are differentially affected by lesions involving afferents of the main and the accessory olfactory systems. The results of lesions that involve the main olfactory system suggest that it provides information primarily concerning those volatile components of HVD that normally signal the presence of a female. Lesions restricted to the accessory olfactory system do not impair a male's interest in or relative preference for HVD but do significantly interfere with subsequent stages of sexual behavior by reducing the amount of mating obtained upon exposure to HVD. Stimulation of the accessory olfactory system by nonvolatile components of HVD has thus been implicated in the production of mating behavior. In these experiments, male hamsters were attracted to female odor and engaged in significant amounts of mating behavior with surrogate females when only the volatile components of HVD were available to them. These behaviors were further enhanced when both volatile and nonvolatile components of HVD were provided. Lesion studies of the afferents involved reinforce the hypothesis that the main olfactory system is preferentially involved with processing those volatile chemical signals in HVD that denote female attractiveness whereas the accessory olfactory system is preferentially involved with processing those chemical signals, both volatile and nonvolatile, that evoke subsequent steps in male sexual behavior.

Animals↗

Patterns of volatile metabolites and nonvolatile trichothecenes produced by isolates of Stachybotrys, Fusarium, Trichoderma, Trichothecium and Memnoniella.

We reported previously that trichodiene, a volatile trichothecene derivative, was produced by a Stachybotrys isolate, also known to produce highly cytotoxic, non-volatile, macrocyclic trichothecenes (satrotoxins). We investigated the relationship between the production of trichodiene and various non-volatile trichothecenes for several molds. Volatile metabolites were concentrated by adsorption on Tenax TA and analyzed by GC/MS, while non-volatile metabolites were separated by HPLC, derivatized and analyzed by GC/MS. Stachybotrys chartarum isolates producing macrocyclic trichothecenes secreted significantly larger amounts of trichodiene and other sesquiterpenes than isolates which only produced simple trichothecenes. The amounts of secreted trichodiene were relatively small in all cases. With the exception of Memnoniella, which excreted small amounts of sesquiterpenes, the other isolates produced varying amounts of sesquiterpenes, including trichodiene, as well as simple tricothecenes, no detectable trichodiene, but large amounts of griseofulvin derivatives. In Stachybotrys there is apparently a correlation between trichodiene and macrocyclic trichothecene production. In the remaining isolates, there was no simple relationship between trichodiene and non-volatile trichothecene synthesis. Trichodiene is produced in larger amounts by Stachybotrys isolates, which also produce satratoxins, but it will be difficult to utilize this metabolite to detect toxic isolates in buildings due to the relatively small amounts excreted.

Air Pollution, Indoor↗

Modeling effects of moisture content and advection on odor causing VOCs volatilization from stored swine manure.

Two models for evaluating the contents and advection of manure moisture on odor causing volatile organic compounds (VOC-odor) volatilization from stored swine manure were studied for their ability to predict the volatilization rate (indoor air concentration) and cumulative exposure dose: a MJ-I model and a MJ-II model. Both models simulating depletion of source contaminant via volatilization and degradation based on an analytical model adapted from the behavior assessment model of Jury et al. In the MJ-I model, manure moisture movement was negligible, whereas in the MJ-II model, time-dependent indoor air concentrations was a function of constant manure moisture contents and steady-state moisture advection. Predicted indoor air concentrations and inhaled doses for the study VOC-odors of p-cresol, toluene, and p-xylene varied by up to two to three orders of magnitude depending on the manure moisture conditions. The sensitivity analysis of both models suggests that when manure moisture movement exists, simply MJ-I model is inherently not sufficient to represent a more generally volatilization process, which can even become stringent as moisture content increases. The conclusion illustrates how one needs to include a wide variety of manure moisture values in order to fully assess the complex volatilization mechanisms that are present in a real situation.

Air Pollution, Indoor↗

Volatilization of 1,2-dibromo-3-chloropropane (DBCP) from soils.

The volatilization of DBCP from soils, as affected by the soil characteristics and application techniques, was studied in a laboratory experiment. The volatilization rate of DBCP applied in water was higher from sandy and silty loam soils than from clay soil. Water added after DBCP application acted as a soil cover, decreasing the volatilization rate. The results obtained with DBCP application in hexane to air-dry soils, indicate that adsorption could be an important factor in reducing the volatilization losses. Diffusion coefficients were calculated from the volatilization parameters, by using a simplified relationship between volatilization losses and diffusion through soil.

Antinematodal Agents↗

Volatile release from liquids: a comparison of in vivo APCI-MS, in-mouth headspace trapping and in vitro mouth model data.

In-mouth volatile release from flavoured water was followed using atmospheric pressure chemical ionization-mass spectrometry (APCI-MS) or using a hand-held, computer-controlled device based on sequential trapping of flavours on Tenax traps. The present results verify recent in vitro data obtained with a sophisticated, fully computerized mouth model apparatus and confirm its validity for the simulation of in-mouth dynamic volatile release. In-nose APCI-MS measurements showed considerable person-to-person variability in non-trained individuals during drinking due to subconscious control of muscles during swallowing and subsequent breathing. Data showed a 'swallow breath' volume reaching the nasal cavity from the throat, not from the mouth cavity. Flavour enriched air from the mouth was shown to be transported to the nose (via exhalation) immediately after the swallowing event, but the dynamic process of volatile equilibration between residuals of the swallowed liquid and the exhaled air predominantly determined volatile in-nose concentration. Owing to its dynamic character, the process of volatile equilibration and release in the throat upon exhalation should be similar to the in-mouth process studied in the present work. A full mechanical simulation of retronasal volatile transport, however, will remain difficult.

Beverages↗

Volatile substances from larval habitats mediate species-specific oviposition in Anopheles mosquitoes.

Oviposition site selection has been recognized as critical both for the survival and population dynamics of mosquitoes. Volatile substances released from larval habitats have been implicated as potential olfactory cues mediating oviposition. In our continuing studies of cues involved in oviposition site selection, we collected material from the larval habitats of Anopheles albimanus Wiedemann and Anopheles vestitipennis Dyar & Knab, i.e., cyanobacterial mats and Typha domingensis Pers. litter, respectively. The volatile compounds were extracted by freeze-drying the material and trapping the volatilized material on a -55 degrees C titanium condenser. For oviposition trials conducted with wild-caught females, the tested volatile materials were pipetted onto filters floating on the surface of distilled water in Teflon beakers that were placed within oviposition cages. For both species, volatile materials in low concentrations increased oviposition, assessed as egg density, whereas there was a shift to reduced oviposition at higher concentrations. Volatile effect was strongly habitat/species-specific as shown by reciprocal treatment tests.

Animals↗

Dietary conjugated linoleic acid with fish oil alters yolk n-3 and trans fatty acid content and volatile compounds in raw, cooked, and irradiated eggs.

We investigated the effect of dietary conjugated linoleic acid (CLA) along with n-3 polyunsaturated fatty acid (n-3 PUFA) on yolk fatty acid composition and volatile compounds in eggs that were raw (RA), hard-boiled (HB), or hard-boiled, irradiated (HBI, 2.5 kGy). Single Comb White Leghorn laying hens (n = 40) were randomly assigned to one of the four experimental diets containing 0, 0.5, 1.0, or 2.0% CLA. Menhaden oil was used as the source of n-3 PUFA. Eggs collected after 6 wk of feeding were analyzed for fatty acids and volatile compounds. The content of docosahexaenoic acid (C22:6 n-3) was reduced (P < 0.05) in eggs from hens fed the 2.0% CLA diet. Eggs from hens fed 0.5% CLA incorporated the highest concentration of docosahexaenoic acid (P < 0.05) with a concomitant reduction in arachidonic acid (P < 0.05). The yolk contents of cis-9 trans-11 CLA and trans-10 cis-12 CLA increased linearly (P < 0.05) as the dietary CLA supply increased. Total monounsaturates were reduced (P < 0.05) with an increase in saturates in yolk. No difference was observed in the total PUFA content of eggs. Total volatiles were reduced in RA eggs from 1.0 and 2.0% CLA diets. 2-Propanone, hexane, and methyl cyclopentane were the major volatiles in RA eggs and were reduced by dietary CLA at 1.0 and 2.0%. Acetaldehyde, pentane, propanol, acetic acid methyl ester, acetic acid ethyl ester, propionic acid methyl ester, 2-methylmethyl propionic acid, 2-propanone, and octane were the major volatiles in HB eggs and were reduced by 2.0% CLA (P < 0.05). No difference was observed in the acetaldehyde, pentane, propanol, acetic acid ethyl ester, octane, or total volatile content of HBI eggs.

Animals↗

Off-odor volatiles and pink color development in precooked, irradiated turkey breast during frozen storage.

The effect of irradiation on color, lipid oxidation, and volatile production of precooked, irradiated turkey breast during frozen storage was studied. Turkey breast muscles were precooked, aerobically or vacuum-packaged, irradiated at 0, 2.5, or 5.0 kGy using a linear accelerator (electron beam), and then frozen-stored at -40 C. Lipid oxidation, volatiles, color values, gas production, and oxidation-reduction potential (ORP) of meat were determined during 3-mo storage periods. Ionizing radiation produced characteristic off-odor volatiles (dimethyldisulfide and methylthioethane) and lipid oxidation products in precooked, frozen turkey breast. The production of volatiles was accelerated by the confounding effect of high irradiation dose, aerobic packaging, and increased storage time. Volatile production and color changes in irradiated, precooked turkey breast were induced by different mechanisms. Irradiation increased pink color in precooked, vacuum-packaged turkey breast, and the pink color was stable during frozen storage. Decreased ORP and increased CO in irradiated meat indicated that denatured CO-heme pigments could be responsible for the pink color in precooked, irradiated turkey breast. Vacuum packaging was better than aerobic packaging in preventing lipid oxidation and oxidation-dependent volatile production, but pink color in precooked, irradiated turkey breast during frozen storage was maintained.

Animals↗

Bacterial volatiles induce systemic resistance in Arabidopsis.

Plant growth-promoting rhizobacteria, in association with plant roots, can trigger induced systemic resistance (ISR). Considering that low-molecular weight volatile hormone analogues such as methyl jasmonate and methyl salicylate can trigger defense responses in plants, we examined whether volatile organic compounds (VOCs) associated with rhizobacteria can initiate ISR. In Arabidopsis seedlings exposed to bacterial volatile blends from Bacillus subtilis GB03 and Bacillus amyloliquefaciens IN937a, disease severity by the bacterial pathogen Erwinia carotovora subsp. carotovora was significantly reduced compared with seedlings not exposed to bacterial volatiles before pathogen inoculation. Exposure to VOCs from rhizobacteria for as little as 4 d was sufficient to activate ISR in Arabidopsis seedlings. Chemical analysis of the bacterial volatile emissions revealed the release of a series of low-molecular weight hydrocarbons including the growth promoting VOC (2R,3R)-(-)-butanediol. Exogenous application of racemic mixture of (RR) and (SS) isomers of 2,3-butanediol was found to trigger ISR and transgenic lines of B. subtilis that emitted reduced levels of 2,3-butanediol and acetoin conferred reduced Arabidopsis protection to pathogen infection compared with seedlings exposed to VOCs from wild-type bacterial lines. Using transgenic and mutant lines of Arabidopsis, we provide evidence that the signaling pathway activated by volatiles from GB03 is dependent on ethylene, albeit independent of the salicylic acid or jasmonic acid signaling pathways. This study provides new insight into the role of bacteria VOCs as initiators of defense responses in plants.

Arabidopsis↗

Effects of feeding Spodoptera littoralis on lima bean leaves. II. Continuous mechanical wounding resembling insect feeding is sufficient to elicit herbivory-related volatile emission.

Herbivore feeding elicits defense responses in infested plants, including the emission of volatile organic compounds that can serve as indirect defense signals. Until now, the contribution of plant tissue wounding during the feeding process in the elicitation of defense responses has not been clear. For example, in lima bean (Phaseolus lunatus), the composition of the volatiles induced by both the insect caterpillar Spodoptera littoralis and the snail Cepaea hortensis is very similar. Thus, a mechanical caterpillar, MecWorm, has been designed and used in this study, which very closely resembles the herbivore-caused tissue damage in terms of similar physical appearance and long-lasting wounding period on defined leaf areas. This mode of treatment was sufficient to induce the emission of a volatile organic compound blend qualitatively similar to that as known from real herbivore feeding, although there were significant quantitative differences for a number of compounds. Moreover, both the duration and the area that has been mechanically damaged contribute to the induction of the whole volatile response. Based on those two parameters, time and area, which can replace each other to some extent, a damage level can be defined. That damage level exhibits a close linear relationship with the accumulation of fatty acid-derived volatiles and monoterpenes, while other terpenoid volatiles and methyl salicylate respond in a nonlinear manner. The results strongly suggest that the impact of mechanical wounding on the induction of defense responses during herbivore feeding was until now underestimated. Controlled and reproducible mechanical damage that strongly resembles the insect's feeding process represents a valuable tool for analyzing the role of the various signals involved in the induction of plant defense reactions against herbivory.

Animals↗

The identification of potential aeroallergen/irritant(s) from oilseed rape (Brassica napus spp. oleifera): volatile organic compounds emitted during flowering progression.

Volatile organic compounds emitted by growing intact oilseed rape plants have been detected using an entrainment apparatus enabling volatile headspace analysis by thermal desorption coupled to capillary gas chromatography-mass spectrometry. In total, 22 volatile compounds were identified as being emitted during the flowering period. The main constituents were alpha-farnesene (a sesquiterpene); beta-myrcene (a monoterpene); linalool (a monoterpene alcohol) and the 'green leaf' volatile (E)-3-hexen-1-ol acetate. These compounds constituted between 50 and 87% (mean 68%) of the total volatiles emitted in all of the entrainments carried out with flowering oilseed rape plants. The remaining constituents consisted of a range of compounds including other terpenoids, the characteristic 'green leaf' volatile (E)-3-hexen-1-ol, short chain alcohols and ketones, organic sulphides and nitrogen-containing compounds. These were generally present as minor constituents but some plant entrainments revealed that higher relative amounts could be emitted. This was particularly apparent for dimethyl disulphide, 3-methyl-2-pentanone, 3-hydroxy-2-butanone, sabinene, isomyrcenol and (E)-3-hexen-1-ol. The possible role of the 22 compounds in respiratory mucosa and conjunctiva irritation associated with airborne releases from oilseed rape is discussed.

Air Pollutants↗

Fungal volatilization of arsenic and antimony and the sudden infant death syndrome.

Fungal volatilization of antimony and other group Vb elements has been proposed to have a causal role in the sudden infant death syndrome (SIDS; cot death). The ability of fungi to produce volatile arsenic and antimony compounds in pure culture was examined using Scopulariopsis brevicaulis, reported as an inhabitant of PVC cot mattress covers, and Phaeolus schweinitzii, a wood decay fungus known to be a good volatilizer of arsenic. Volatile arsenic compounds were detected from all cultures grown on arsenic-supplemented media, but antimony volatilization was not reliably detected. Although antimony levels above the baseline sensitivity of the analytical technique were detected in four (out of 24) of the samples analyzed, the concentrations recorded were too low to be reliably interpreted as evidence for volatilization. Our results are discussed in relation to hypotheses regarding the causes of SIDS.

Antimony↗

Role of volatile fatty acids in development of the cecal microflora in broiler chickens during growth.

It is known that volatile fatty acids can inhibit growth of species of the family Enterobacteriaceae in vitro. However, whether these volatile fatty acids affect bacterial populations in the ceca of chickens is unknown. Therefore, a study was conducted to investigate if changes in volatile fatty acids in ceca of broiler chickens during growth affect bacterial populations. Results showed that members of the Enterobacteriaceae and enterococci are present in large numbers in 3-day-old broilers and start to decrease when broilers grow older. Lactobacilli are present in large numbers as well in 3-day-old broilers, but they remain stable during the growth of broilers. Acetate, butyrate, and propionate increase from undetectable levels in 1-day-old broilers to high concentrations in 15-day-old broilers, after which they stabilize. Significant negative correlations could be calculated between numbers of Enterobacteriaceae and concentrations of undissociated acetate, propionate, and butyrate. Furthermore, pure cultures of Enterobacteriaceae isolated from the ceca were grown in the presence of volatile fatty acids. Growth rates and maximal optical density decreased when these strains grew in the presence of increasing volatile fatty acid concentrations. It is concluded that volatile fatty acids are responsible for the reduction in numbers of Enterobacteriaceae in the ceca of broiler chickens during growth.

Animals↗

Volatile fatty acid findings in vaginal fluid compared with symptoms, signs, other laboratory results, and susceptibility to tinidazole of malodorous vaginal discharges.

The relevance of volatile fatty acids as a diagnostic test in 79 women with abnormal vaginal discharge was evaluated by a blind, randomised, and placebo controlled trial of tinidazole as a single oral 2 g dose. Automated gas chromatography of ether extracts of discharges taken before treatment showed volatile fatty acids in 18. Volatile fatty acids correlated with malodorous, colour, and microscopically assessed altered bacterial flora and clue cells. At follow up one week later, the odour, colour, and volatile fatty acids in the vaginal discharge of women treated with tinidazole had become normal more often than in those receiving placebo. The disappearance of volatile fatty acids correlated with clinically assessed improvement in women treated with tinidazole. The volatile fatty acid test as an indicator of anaerobic bacterial flora is objective, technically simple and fast, has few problems of sample size and transportation, and may be useful in the aetiological classification and follow up treatment of non-specific vaginal discharges.

Body Fluids↗