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Practical approaches to plant volatile analysis.

Plants emit volatile organic compounds (VOCs) that play important roles in their interaction with the environment and have a major impact on atmospheric chemistry. The development of static and dynamic techniques for headspace collection of volatiles in combination with gas chromatography-mass spectrometry analysis has significantly improved our understanding of the biosynthesis and ecology of plant VOCs. Advances in automated analysis of VOCs have allowed the monitoring of fast changes in VOC emissions and facilitated in vivo studies of VOC biosynthesis. This review presents an overview of methods for the analysis of plant VOCs, including their advantages and disadvantages, with a focus on the latest technical developments. It provides guidance on how to select appropriate instrumentation and protocols for biochemical, physiological and ecologically relevant applications. These include headspace analyses of plant VOCs emitted by the whole organism, organs or enzymes as well as advanced on-line analysis methods for simultaneous measurements of VOC emissions with other physiological parameters.

Gas Chromatography-Mass Spectrometry↗

Characterization of a BAHD acyltransferase responsible for producing the green leaf volatile (Z)-3-hexen-1-yl acetate in Arabidopsis thaliana.

Green-leaf volatiles are commonly emitted from mechanically and herbivore-damaged plants. Derived from the lipoxygenase pathway, these compounds may serve as attractants to predators and parasitoids of herbivores, prevent the spread of bacteria and fungi, and induce several important plant defense pathways. In Arabidopsis thaliana, the major volatile released upon mechanical wounding of the leaves is the GLV ester, (Z)-3-hexen-1-yl acetate. We have characterized a member of the BAHD acyltransferase gene family At3g03480 which catalyzes the formation of (Z)-3-hexen-1-yl acetate from acetyl-CoA and (Z)-3-hexen-1-ol. The encoded acetyl CoA:(Z)-3-hexen-1-ol acetyltransferase (CHAT) has the ability to accept several medium-chain-length aliphatic and benzyl-derived alcohols, but has highest catalytic efficiency with (Z)-3-hexen-1-ol. The highest expression of CHAT occurs in the leaves and stems. Upon mechanical damage, the (Z)-3-hexen-1-yl acetate emission peaked after 5 min and declined to pre-damage levels after 90 min. However, CHAT gene transcript levels increased much more slowly with the highest levels detected between 3 and 6 h after wounding. An increase in CHAT enzyme activity in vitro followed the transcript increase, with levels peaking between 10 and 12 h after wounding. Plants expressing either an RNAi cassette for the CHAT gene or plants harboring a T-DNA insertion in the CHAT coding region had greatly reduced (Z)-3-hexen-1-yl acetate emission, showing that the CHAT enzyme is responsible for the in planta production of this ester.

Acetates↗

In vitro screening for metabolic interactions among frequently occurring binary mixtures of volatile organic chemicals in Norwegian occupational atmosphere.

Surveys of Norwegian industrial occupational atmosphere between 1983 to 1996, have identified the 12 most frequent occurring binary combinations of volatile organic chemicals. These combinations were tested in vitro for mutual inhibition or enhancement of metabolism by the head space vial equilibration technique with liver S9 obtained from in vivo untreated or pretreated (with the binary mixture) rats. The in vitro system responded to in vivo pretreatment by increasing the metabolic rate of several potentially toxic organic chemicals such as toluene, xylene, styrene, and dichloromethane. In untreated liver S9, the metabolism of several of the tested binary pairs was inhibited when coexposed in vitro to their most prevalent follower as shown for instance for ethanol (with ethyl acetate), dichloromethane (with styrene) and mutually between toluene and xylene. This inhibitory effect disappeared, however, for several of the solvents when combined with the in vivo induced liver S9, a situation which may be the most relevant for occupational exposure. It is concluded that several metabolic interactions occur between low-molecular weight volatile chemicals found in occupational air. These are both inductive and inhibitory in nature and a further mechanistic evaluation including a higher number of differentiated dosage levels, must be performed before a possible health hazard can be confirmed or rejected for the investigated combinations.

Air Pollutants, Occupational↗

Volatile signaling in plant-plant interactions: "talking trees" in the genomics era.

Plants may "eavesdrop" on volatile organic compounds (VOCs) released by herbivore-attacked neighbors to activate defenses before being attacked themselves. Transcriptome and signal cascade analyses of VOC-exposed plants suggest that plants eavesdrop to prime direct and indirect defenses and to hone competitive abilities. Advances in research on VOC biosynthesis and perception have facilitated the production of plants that are genetically "deaf" to particular VOCs or "mute" in elements of their volatile vocabulary. Such plants, together with advances in VOC analytical instrumentation, will allow researchers to determine whether fluency enhances the fitness of plants in natural communities.

Adaptation, Physiological↗

The molecular basis for wine grape quality--a volatile subject.

Volatile organic compounds are important flavor components of finished wines. In addition to winemaking practices, which shape wine quality, cultivation of the grape berries in the vineyard each season affects the production of volatile organic compounds as well as other chemical components that ultimately contribute to our perception of flavor in finished wines. By studying how berry flavor components are determined by the interplay of vine genotypes, the environment, and cultivation practices at the molecular level, scientists will develop advanced tools and knowledge that will aid viticulturalists in consistently producing balanced, flavorful berries for wine production.

Agriculture↗

Parent volatiles in comet 9P/Tempel 1: before and after impact.

We quantified eight parent volatiles (H2O, C2H6, HCN, CO, CH3OH, H2CO, C2H2, and CH4) in the Jupiter-family comet Tempel 1 using high-dispersion infrared spectroscopy in the wavelength range 2.8 to 5.0 micrometers. The abundance ratio for ethane was significantly higher after impact, whereas those for methanol and hydrogen cyanide were unchanged. The abundance ratios in the ejecta are similar to those for most Oort cloud comets, but methanol and acetylene are lower in Tempel 1 by a factor of about 2. These results suggest that the volatile ices in Tempel 1 and in most Oort cloud comets originated in a common region of the protoplanetary disk.

Jupiter↗

Human vaginal secretions: volatile fatty acid content.

Vaginal samples (682) were collected by a tampon method from 50 healthy young women. Samples were analyzed by gas chromatography. The volatile aliphatic acids increased during the late follicular phase of the menstrual cycle and declined progressively during the luteal phase. Women on oral contraceptives had lower amounts of volatile acids and did not show any rhythmic changes in acid content during the menstrual cycle. These same substances possess sex-attractant properties in other primate species.

Acetates↗

Effects of acidification on mercury methylation, demethylation, and volatilization in sediments from an acid-susceptible lake.

The effect of experimental acidification on mercury methylation, demethylation, and volatilization was examined in surficial sediment samples from a weakly buffered northern Wisconsin lake. All mercury transformations were measured with radioisotopic tracers. Acidification of sediment pH with H2SO4, HCl, or HNO3 significantly decreased 203Hg(II) methylation. Acidification of pH 6.1 (ambient) sediments to pH 4.5 with either H2SO4 or HCl inhibited methylation by over 65%. The decreased methylation was due to the increased hydrogen ion concentration because methylation was not affected by concentrations of Na2SO4 or NaCl equimolar to the amount of acid added. Inhibition of methylation was observed even after prolonged acidification of sediments to pH 5.0 for up to 74 days. Acidification of sediments to pH 5.5, 4.5, and 3.5 with HNO3 resulted in a near complete inhibition of methylation at each pH. Similarly, the addition of equimolar amounts of NaNO3 resulted in a near complete inhibition of methylation, indicating that the inhibition was due to the nitrate ion rather than to the acidity. Demethylation of methyl mercury was not affected by pHs between 8.0 and 4.4, but sharply decreased below pH 4.4. Volatilization of 203Hg(II) from surface sediments was less than 2% of methylation activity and was not significantly different from that in killed sediments. This study indicated that acidification of sediments inhibits mercury methylation and that the observed increase in the mercury burdens in fish from low pH lakes is not due to increased production of methylmercury in sediments.

Bacteria↗

Antibacterial action of essential oils of Artemisia as an ecological factor. II. Antibacterial action of the volatile oils of Artemisia tridentata (big sagebrush) on bacteria from the rumen of mule deer.

Rumen microorganisms of wild and captive deer were subjected to increasing amounts of volatile oils. The oils had a marked antibacterial effect on the rumen bacteria when the concentration reached approximately 16 muliters of oil per 10 ml of rumen fluid nutrient broth. The gross reactions of rumen bacteria obtained from wild, as well as captive, deer to the volatile oils seemed to be of the same magnitude; thus no adaptation by the bacteria to the oils was apparent.

Animals↗

Predictive value of the "clue cells" investigation and the amine volatilization test in vaginal infections caused by Gardnerella vaginalis.

Although still controversial, an etiologic role of Gardnerella vaginalis is imputed in vaginitis. Besides isolation of the organism by culture, two alternative diagnostic procedures have been claimed to be useful: the investigation of "clue cells" in clinical specimens and the amine volatilization test or fishy odor perception in genital secretions. Herein we report on the findings of the simultaneous use of G. vaginalis isolation, the clue cell test and amine volatilization perception in specimens from 1,263 consecutive female patients referred to our clinic. Our results show that the simultaneous use of both alternative tests is very useful as a screening procedure. A negative result of both tests predicts a negative culture result in 99% of the cases. However, a positive result of either or both should be considered as an indication to proceed to culture and not as diagnostic of infection.

Adolescent↗

Detection of alcohols and volatile fatty acids by head-space gas chromatography in identification of anaerobic bacteria.

A head-space gas chromatographic technique for the analysis of volatile bacterial metabolites is described. Bacteroides fragilis, Clostridium perfringens, and Propionibacterium acnes, cultured in a glucose-containing peptone yeast extract medium, were studied. The head-space technique was compared with the injection of the complete liquid culture medium, and solvent extracts thereof, into the gas chromatograph. Volatile fatty acids could be detected by all three methods, whereas alcohols produced by C. perfringens and P. acnes were detectable only in the head-space chromatograms. Both FFAP and Porapak Q were used as gas chromatography stationary phase. Porapak Q was found more suitable than FFAP for the separation of alcohols. The head-space technique requires a minimum of preparation before the analysis and is well suited to automation.

Alcohols↗

Gas-liquid chromatographic analysis of synovial fluid: volatile short-chain fatty acids in septic arthritis.

Volatile short-chain fatty acids (SCFA) in synovial fluid from 80 patients were quantified by gas-liquid chromatography (GLC). Characteristic patterns of volatile SCFA could not be associated with septic, nonseptic inflammatory, or noninflammatory groups. Mean concentrations of pentanoic and hexanoic acids were similar in all groups studied. In the septic arthritis group 3 of 4 patients with acetic acid and of 3 of 3 with 3-methyl butanoic acid had culture-proved staphylococcal infections. In patients with synovial fluid findings consistent with septic arthritis, including markedly raised leucocyte count, decreased glucose level, or detectable succinic acid, the detection of acetic and 3-methyl butanoic acids by GLC analysis may increase the suspicion that Staphylococcus aureus is the cause of the septic arthritis.

Arthritis↗

Effect of monensin sodium on the performance and proportions of rumen volatile fatty acids of Friesian bulls.

The effect of including monensin sodium at a dose rate of 40 mg/kg in the diet of Friesian bulls (mean initial liveweight 158 kg) fed a barley-based diet ad libitum was assessed in terms of growth rate, feed intake, feed and energy conversion efficiency and the concentration and molar proportions of ruminal volatile fatty acids over a 157-day period. Mean daily feed dry matter intake for the treated group (5.4 kg) was 24 per cent lower than the control group (7.1 kg). The treated group grew at a significantly slower rate than the control animals although feed conversion efficiency was improved by the treatment. A comparison with the results of other workers suggested that the dose rate used had been too high. There were no significant differences in the molar proportions of volatile fatty acids in rumen fluid as a result of monensin usage. While many workers have shown increases in the molar proportions of propionic acid the fact that the control group in this experiment had a very high value (48 per cent) may explain the lack of response.

Administration, Oral↗

Effects of ammonia and volatile fatty acids produced by oral bacteria on tissue culture cells.

Culture filtrates of several bacterial species isolated from the oral cavity were tested for their effects on two types of tissue culture cells: Vero cells, the continuous cell line of African green monkey kidney cells; and chondrocytes, isolated from 15-day-old chick embryo tibiae. Only a limited number of bacterial species--i.e., the asaccharolytic black-pigmented Bacteroides species and Fusobacterium species--affected the two cell types. The effect on Vero cells, detected by the rounding of the cells, correlated with the butyric acid concentration in the bacterial supernatant, which confirms previous findings. A small enhancement of this effect was found with propionic acid and ammonium ions. The same strains which affected Vero cells also affected chondrocytes, detected by a vacuolization of the cells. However, volatile fatty acids on their own had no visible effect on these cells. Instead, ammonium ion in the culture filtrate, when present in concentrations of 20 to 60 mmol/L, proved to be responsible for vacuolization of the chondrocytes. The volatile fatty acids (butyric and propionic) had a limited additive effect. No effects were visible with cell extracts of the bacteria.

Actinomyces↗

Determinants of volatile general anesthetic potency: a preliminary three-dimensional pharmacophore for halogenated anesthetics.

We investigated the molecular basis for the immobilizing activity of halogenated volatile anesthetics using comparative molecular field analysis. In vivo potency data (expressed as minimum alveolar concentrations) for 69 structurally diverse anesthetics were obtained from the literature. The drugs were randomly divided into a training set (n = 52) used to derive the activity model and a test set (n = 17) used to independently assess the model's predictive power. The anesthetic structures were aligned so as to maximize their similarity in molecular shape and electrostatic potential to the most potent drug in the group, CF2H-(CF2)3-CH2OH. The conformers and alignments of the anesthetics with maximum similarity (calculated as Carbo indices) were retained and used to derive the comparative molecular field analysis models. The final model explained 94.2% of the variance in the observed activities of the training set compounds. The model showed good predictive capability for both the training set (cross-validated r2 = 0.705) and randomly excluded test set anesthetics (r2 = 0.837). Three-dimensional pharmacophoric maps were derived to identify the spatial distribution of key areas where steric and electrostatic interactions are important in determining immobilizing activity of the halogenated drugs and were compared with our previously published maps obtained for nonhalogenated volatile anesthetics.

Anesthetics, General↗