A gas chromatographic investigation of the influence of different carbon sources on the production of volatile compounds by Dipodascus aggregatus

Summary The yeast fungus Dipodascus aggregatus was cultivated aerobically on different carbon sources. The growth was measured turbidimetrically and related to the simultaneus production of volatile compounds, which was determined by a gas chromatographic head-space technique. The same culture could be analyzed many times and up to 15 components were detected in a chromatogram recorded in 18 min. The main peaks were identified and the chromatograms quantitatively evaluated by peak height measurements.All the carbon sources tested were utilized for growth even if the lag phase was prolonged on xylose and ethanol. The production of volatile compounds from the different carbon sources decreased as follows; ethanol>glucose>fructose>glycerol >xylose and succinic acid. A good carbon source for growth could be unsuitable for the formation of volatile products and vice versa.The time course production of volatile components was recorded. On each separate carbon source the formation of volatiles was correlated to growth until the end of the exponential phase of growth.An extraction procedure including the addition of an internal standard was used to determine the exact concentrations of 9 components at near optimum production. The amounts varied between 0.1–32 mg/l medium when the fungus was grown on 5% glucose.     

GC-MS SPME profiling of rhizobacterial volatiles reveals prospective inducers of growth promotion and induced systemic resistance in plants

Chemical and plant growth studies of Bacilli strains GB03 and IN937a revealed that the volatile components 2,3-butanediol and acetoin trigger plant growth promotion in Arabidopsis. Differences in growth promotion when cytokinin-signaling mutants are exposed to GB03 versus IN937a volatiles suggest a divergence in chemical signaling for these two bacterial strains. To provide a comprehensive chemical profile of bacterial volatiles emitted from these biologically active strains, headspace solid phase microextraction (SPME) coupled with software extraction of overlapping GC-separated components was employed. Ten volatile metabolites already reported from GB03 and IN937a were identified as well as 28 compounds not previously characterized. Most of the newly identified compounds were branched-chain alcohols released from IN937a, at much higher levels than in GB03. Principal component analysis clearly separated GB03 from IN937a, with GB03 producing higher amounts of 3-methyl-1-butanol, 2-methyl-1-butanol and butane-1-methoxy-3-methyl. The branched-chain alcohols share a similar functional motif to that of 2,3-butanediol and may afford alternative structural patterns for elicitors from bacterial sources.     

Shoyu (Soy Sauce) Flavor Components: Neutral Fraction

From the vacuum distilled volatiles of shoyu, a neutral fraction was obtained. Shoyu was also directly extracted with dichloromethane and then the extract was separated into ten (A~J) fractions. The J fraction was a neutral one. The J fraction was further separated into twelve fractions by liquid column chromatography. All fractions obtained were analyzed by gas chromatography and combined gas chromatography-mass spectrometry. Consequently, 142 components were identified, 82 of which have not been reported previously as volatile constituents of shoyu. The identified compounds were 37 hydrocarbons, 22 alcohols, 22 carbonyls, 22 esters, 12 furans, 6 sulfurous compounds, 1 pyrone, 5 phenols, 1 furanone, 1 acid, 1 lactone and 12 other compounds. From the results of quantitative analysis and organoleptic evaluation, phenylacetal-dehyde is considered to be most important in the neutral fraction.     

Antimicrobial volatile organic compounds affect morphogenesis-related enzymes in Guignardia citricarpa,causal agent of citrus black spot

Although non-volatile substances toxic to plant pathogenic microorganisms have been extensively studied over the years, few studies have focused on microbial volatile organic compounds (VOCs). The VOCs produced by the yeast Saccharomyces cerevisiae strain CR-1, used in fermentative processes for fuel ethanol production, are able to inhibit the vegetative development of the fungus Guignardia citricarpa, causal agent of the disease citrus black spot. How microbial VOCs affect the development of fungi is not known. Thus, the objective of the present work was to study the effect of the artificial mixture of VOCs identified from S. cerevisiae on intracellular enzymes involved in the mycelial morphogenesis in G. citricarpa. The phytopathogenic fungus was exposed to artificial mixture of VOCs constituted by alcohols (ethanol, 3-methyl-1-butanol, 2-methyl-1-butanol and phenylethyl alcohol) and esters (ethyl acetate and ethyl octanoate) in the proportions naturally found in the atmosphere produced by the yeast. The VOCs inhibited considerably the mycelial development and interfered negatively with the production of the morphogenesis-related enzymes. After 72 h of exposure to the VOCs the laccase and tyrosinase activities decreased 46 and 32%, respectively, however, the effect on the chitinase and β-1,3-glucanase activities was lower, 17 and 13% of inhibition, respectively. Therefore, the exposure of the fungus to the antimicrobial volatiles can influence both fungal mycelial growth rate and activity of enzymes implicated in morphogenesis. This knowledge is important to understand the microbial interactions mediated by VOCs in nature and to develop new strategies to control plant pathogens as G. citricarpa in postharvest.     

Dynamic changes in volatile emissions of breeding burying beetles

Burying beetles reproduce on small vertebrate carcasses by exhibiting elaborate biparental brood care. Partner recognition in breeding Nicrophorus species (Coleoptera: Silphidae) relies substantially on information encoded in cuticular hydrocarbon profiles. Until recently, it was unknown whether breeding burying beetles also produce volatile low molecular weight substances and, if so, which functions can be attributed to such volatiles. The present study reports a survey of the volatiles released by males and females of Nicrophorus vespilloides Herbst in nonbreeding status and at different stages of breeding. Headspace analyses are performed by using solid phase micro‐extraction fibres and gas chromatography–mass spectrometry. The volatiles released by nonbreeding males and females include phenolic compounds, alcohols, aldehydes and ketones and are quite similar in both sexes. With the onset of breeding, the volatile profiles of males and females become distinct, with a number of female‐specific compounds occurring. An analysis of the anal secretions reveals the presence of some of the compounds previously detected in the headspace analysis. The specific chemical properties suggest that some of the volatiles may function against competitors and parasites, such as bacteria, fungi, nematodes and arthropods at the carcass breeding resource. By contrast, the emission of 4‐methyl branched esters by the females closely parallels the emission of the terpenoid methyl geranate and they may function together as a complex signal by the females. Signalling traits associated with biparental care and specific constraints associated with the ephemeral nature of the breeding resource may explain the occurrence of both groups of compounds in the volatile profiles.     

Comprehensive volatile organic compounds profiling of Bacillus species with biocontrol properties by head space solid phase microextraction with gas chromatography-mass spectrometry

The eight Bacillus strains, used as biocontrol agents with proven antagonistic effect against plant pathogens, produced antifungal volatile organic compounds (VOCs). Bioassay in sealed dishes revealed that the VOCs from each Bacillus strain significantly inhibited the mycelial growth (56–82%) of Fusarium solani. The effective antifungal VOCs were extracted using headspace solid phase microextraction and further identified by gas chromatography-mass spectrometry technique. The detected volatile compounds could be chemically grouped into ketones, alcohols, aldehydes, pyrazines, acids, esters, pyridines and benzene compounds. The ketones and alcohols were predominant in the VOCs from eight Bacillus strains whereas the ketones, including 3-methyl-2-pentanone, 2-heptanone, 2-octanone, 2-decanone, 5-methyl-2-hexanone, 2-nonanone, 2-dodecanone, 2-undecanone, 5-methyl-2-heptanoneand2-pentanone, were the most common and principal components in all strains. Present results showed that the eight Bacillus strains are rich resources of bioactive volatiles, which may play an important role in the inhibition on F. solani. Studies are under the way to determine effects of those compounds against plant pathogens and to find the possible action mechanisms.     

Morphogenetic effects of some volatile,organic compounds on Pestalotia rhododendri

Summary Some volatile alcohols, acids, esters, aldehydes and ketones influenced the morphogenesis of Pestalotia rhododendri when added through the gas phase. Linear growth, formation of aerial hyphae and production of conidial spores were determined. Mostly, increasing amounts of test substance inhibited linear growth and only one case of stimulation was noticed, viz. by nonenal.Many substances stimulated formation of aerial hyphae, particularly allyl alcohol, acetaldehyde and nonanal, but inhibitions were more common.Spore production was promoted by compounds from all the chemical classes investigated. The activity decreased as follows; alcohols > esters > acids, aldehydes > ketones. Inhibitions of spore production were also observed.The effects were reproducible on two different media. Pestalotia sydowiana exhibited the same main responses as the other Pestalotia species, but only two substances, furfural and n-propanol, were observed to promote spore formation of this species.     

细菌产生的挥发性物质及其生物学功能

细菌产生的挥发性物质种类繁多,成分复杂,是寻找具有特殊生物学功能的天然化合物的一个重要来源。近些年,越来越多的研究人员关注细菌产生的挥发性物质及其生物学功能。概述了细菌产生的挥发性物质的种类以及分析鉴定挥发性物质成分的方法;着重评述了细菌产生的挥发性物质对真菌、细菌、动物和植物生长和代谢的影响,并对细菌产生的挥发性物质未来的主要研究方向进行了展望。     

Study of the volatile compounds produced by Debaryomyces hansenii NRRL Y-7426 during the fermentation of detoxified concentrated distilled grape marc hemicellulosic hydrolysates

The volatile compounds produced by Debaryomyces hansenii NRRL Y-7426 during the fermentation of detoxified concentrated distilled grape marc hemicellulosic hydrolysates was analysed by GC?CMS. Thirty-five compounds corresponding to ten groups of volatile compounds: terpenes, higher alcohols, C₆ alcohols, aldehydes, volatile acids, acetates, ethyl esters, volatile phenols, sulphur compounds and hydrocarbons were identified. The supplementation with commercial nutrients increased the concentration of 2-phenylethanol, a commercial flavour and fragrance compound, with a rose-like odour, employed in cosmetics and food industries; and other positive compounds to the aroma such as terpenes and ethyl esters. Non-supplemented media produced the highest content in higher alcohols, volatile acids, sulphur compounds and in the majority of hydrocarbons detected, meanwhile supplementation with vinasses hardly produced volatile compounds. Only four volatile compounds contributed directly to the aroma according to the OAVs values higher than 1. Finally, a PCA analysis allowed accounting for 100?% of the variance.     

Volatile emission of different plant parts and fruit development from Italian cherry plums (Prunus cerasifera and P. cerasifera ‘Pissardii’)

The chemical composition of volatiles emitted in vivo from different plant parts of P. cerasifera and P. cerasifera ‘Pissardii’ were collected during the entire biological cycle of the plant growth and analyzed by gas chromatography mass spectrometry (GC-MS) after solid phase micro-extraction (SPME). All the data were submitted to multivariate statistical analysis evidencing many differences amongst the selected plant parts and growth stages. A total of 136 compounds were identified corresponding to 90.1–99.6% of the whole aroma profile of cherry plum samples. Non-terpenes were the most abundant class of constituents present in the volatile emission of all analyzed samples. In particular, the aroma of both fruit stages was mainly characterized by alcohols and esters in different amounts.     

Temporal relationship between emitted and endogenous floral scent volatiles in summer‐ and winter‐blooming Jasminum species

Jasminum spp. is cultivated for their fragrant flowers used in essential oil production and cosmetic uses. An attempt was made to study the temporal variations in floral scent volatiles composition including emitted, free endogenous and glycosyl‐linked volatile compounds from two summer‐blooming species namely, Jasminum auriculatum and Jasminum grandiflorum as well as from two winter‐blooming species namely, Jasminum multiflorum and Jasminum malabaricum. The overall emitted volatile organic compounds (VOCs) were found to be highest when the matrix Porapak Q 80/100 was used with dichloromethane (DCM) as elution solvent. The floral volatile emission from bud to senescence exhibited nocturnal maxima pattern for both the summer‐blooming species. Both the winter‐blooming species emitted its highest concentration at noon. The free endogenous concentrations of all VOCs were low when corresponding emitted concentrations were high. Enzymatic treatment of petal extract revealed that several aromatic volatiles including aromatic alcohols and monoterpenols are synthesized and stored in the flowers as water‐soluble glycosides; these compounds were shown to accumulate in higher amounts in flowers at late bud stage. These findings indicate the utilization of the precursors, i.e. the volatile‐conjugates, through hydrolysis followed by their release as free‐volatiles at flower opening stage. The outcome as a whole suggests a linkage among the temporal pattern of emitted volatiles, free‐endogenous volatiles and glycoside‐bound volatile compounds in all above studied Jasminum spp. and provided an overview of their floral volatilome.     

Composition and emission dynamics of migratory locust volatiles in response to changes in developmental stages and population density

Chemical communication plays an important role in density‐dependent phase change in locusts. However, the volatile components and emission patterns of the migratory locust, Locusta migratoria, are largely unknown. In this study, we identified the chemical compositions and emission dynamics of locust volatiles from the body and feces and associated them with developmental stages, sexes and phase changes. The migratory locust shares a number of volatile components with the desert locust (Schistocerca gregaria), but the emission dynamics of the two locust species are significantly different. The body odors of the gregarious nymphs in the migratory locust consisted of phenylacetonitrile (PAN), benzaldehyde, guaiacol, phenol, aliphatic acids and 2,3‐butanediol, and PAN was the dominant volatile. Volatiles from the fecal pellets of the nymphs primarily consist of guaiacol and phenol. Principal component analysis (PCA) showed significant differences in the volatile profiles between gregarious and solitary locusts. PAN and 4‐vinylanisole concentrations were significantly higher in gregarious individuals than in solitary locusts. Gregarious mature males released significantly higher amounts of PAN and 4‐vinylanisole during adulthood than mature females and immature adults of both sexes. Furthermore, PAN and 4‐vinylanisole were completely lost in gregarious nymphs during the solitarization process, but were obtained by solitary nymphs during gregarization. The amounts of benzaldehyde, guaiacol and phenol only unidirectionally decreased from solitary to crowded treatment. Aliphatic aldehydes (C7 to C10), which were previously reported as locust volatiles, are now identified as environmental contaminants. Therefore, our results illustrate the precise odor profiles of migratory locusts during developmental stages, sexes and phase change. However, the function and role of PAN and other aromatic compounds during phase transition need further investigation.     

白星花金龟对寄主植物挥发物的电生理反应

[目的]白星花金龟Protaetia brevitarsis是我国经济上重要的农业害虫.本研究拟鉴定苹果、葡萄和玉米3种果实在两种状态(健康与受白星花金龟成虫为害)下的挥发物中能引起白星花金龟雌成虫触角电生理反应的挥发物成分.[方法]采用动态顶空吸附法收集苹果、葡萄和玉米3种果实的挥发物,用气相色谱-触角电位联用技术(...     

9 种海洋硅藻挥发性成分的比较分析

采用顶空固相微萃取气相色谱一质谱联用技术,通过SIMCA．P分析软件,对硅藻门下处于生长平台后期的9种海洋硅藻的挥发性成分进行比较分析研究。结果表明：在平台后期硅藻的挥发性成分中,醛类物质的含量和种类占据了很大的优势,壬醛、2,4-辛二烯醛、2-己烯醛、2-壬烯醛、2,4-庚二烯醛等是硅藻挥发性成分的主要构成物质,十四烷、十五烷、1-十五烯、十六烷等烷烃类是硅藻中含量仅次于醛类物质的挥发性成分,3,5-辛烯基-3-酮等短链酮类和1-辛烯基-3-醇是硅藻中含量较高的酮类和醇类物质,9种硅藻中均检测二甲基硫的存在。差异性较大的挥发性物质主要有壬醛、8-十七碳烯、1-戊烯．3．酮、1-己烯-3．醇、2,6-二甲基一苯酚,它们决定了不同的硅藻有着各自不同的风味。研究结果表明这些挥发性物质对硅藻的饵料价值、养殖生物的肉质风味、水环境的异味形成有着重要意义。     

Effect of leucine on aroma volatiles production from Ceratocystis fimbriata grown in liquid culture

Aroma volatiles produced by Ceratocystis fimbriata on a defined liquid synthetic medium with and without the addition of leucine were identified by gas chromatography–mass spectrometry and quantified by gas chromatography-flame ionisation detection in the liquid medium as well as in the headspace. Volatiles were extracted from the liquid by simultaneous steam distillation–solvent extraction. Ceratocystis fimbriata produced a complex set of volatile intermediary metabolites, of which ethanol was the dominant compound (92–95% of total volatiles). Low molecular weight esters, alcohols, aldehydes, ketones, alkanes, and carboxylic acids were identified in the liquid broth. Alcohols and esters were the most abundant aroma volatiles. Leucine addition effected further growth and higher volatiles production. In the headspace, ethanol and ethyl acetate accounted for 92% of total volatiles over the synthetic medium and 89% when leucine was added. Aroma perception (fruity and banana) correlated closely with liquid and headspace total volatiles.     

The influence of different nitrogen sources on the production of volatile compounds by Dipodascus aggregatus

Summary The yeast fungus Dipodascus aggregatus was grown aerobically on 9 different nitrogen sources and the production of volatile compounds determined by a gas chromatographic head-space technique. Excellent growth was supported by glutamine, aspartic acid, asparagine, (NH₄)₂-tartrate and NH₄H₂PO₄. Valine, leucine, and particularly isoleucine were utilized with a somewhat lower growth rate. Lysine was rapidly utilized after a prolonged lag phase.The highest production of volatile compounds was obtained from leucine and isoleucine. At least 20 volatile compounds were formed from each of them and many products were detected in high concentrations. Intermediate amounts of volatile compounds were produced from asparagine, the ammonium salts and valine, and low amounts from lysine, glutamine and aspartic acid.Ethyl acetate was a major product irrespective of the nitrogen source used. Regarding the pattern of volatile compounds produced, leucine, isoleucine and valine had much in common. Most of the volatile products formed from these amino acids contained a branched carbon chain and at least three high-boiling components eluted later than n-amyl acetate from the gas chromatographic column. The other six nitrogen sources could be grouped together. In general the same volatile compounds were formed from these sources, but the quantities of the individual compounds differed. Only one component eluted later than n-amyl acetate. No basic difference in production of volatile compounds was observed between the ammonium salts and -amino compounds like lysine and asparagine.     

Volatile Compound-Mediated Interactions between Barley and Pathogenic Fungi in the Soil

Plants are able to interact with their environment by emitting volatile organic compounds. We investigated the volatile interactions that take place below ground between barley roots and two pathogenic fungi, Cochliobolus sativus and Fusarium culmorum. The volatile molecules emitted by each fungus, by non-infected barley roots and by barley roots infected with one of the fungi or the two of them were extracted by head-space solid phase micro extraction and analyzed by gas chromatography mass spectrometry. The effect of fungal volatiles on barley growth and the effect of barley root volatiles on fungal growth were assessed by cultivating both organisms in a shared atmosphere without any physical contact. The results show that volatile organic compounds, especially terpenes, are newly emitted during the interaction between fungi and barley roots. The volatile molecules released by non-infected barley roots did not significantly affect fungal growth, whereas the volatile molecules released by pathogenic fungi decreased the length of barley roots by 19 to 21.5% and the surface of aerial parts by 15%. The spectrum of the volatiles released by infected barley roots had no significant effect on F. culmorum growth, but decreased C. sativus growth by 13 to 17%. This paper identifies the volatile organic compounds emitted by two pathogenic fungi and shows that pathogenic fungi can modify volatile emission by infected plants. Our results open promising perspectives concerning the biological control of edaphic diseases.     

Volatile compounds from Pinus silvestris stimulating the growth of wood-rotting fungi

Summary Certain volatile organic acids stimulated the growth of Stereum sanguinolentum when added through the gas phase or to the liquid nutrient medium. Some of the acids were identified in extract of heat-treated wood of pine (Pinus silvestris), which had been shown (Suolahti, 1951) to have a growth-promoting effect on Stereum sanguinolentum and other wood-rotting fungi.The effect of volatile materials from a heat-treated piece of pine-wood and of caproic acid was tested on 13 other wood-rotting fungi. The growth of 11 of them was stimulated by volatiles from pine-wood, only 2 by caproic acid.The morphology of Coniophora cerebella was altered when volatiles from a neutral fraction of pine-wood extract was added through the gas phase.     

Exudation of low molecular weight compounds (thiobismethane, methyl isocyanide, and methyl isothiocyanate) as a possible chemical defense mechanism in the marine sponge Ircinia felix

The volatile constituents of the marine sponge Ircinia felix were obtained by dynamic headspace extraction and analyzed by HRGC, HRGC-MS and HRGC-Odor at sniffing port. Fifty-nine volatiles were identified for the first time in the odor of this sponge. Hydrocarbons (32.9%), alcohols (17.8%) and carbonyl compounds (16.0%) predominated in the sponge volatile profile, followed by esters (11.6%), halogen compounds (8.6%), ethers (7.7%), nitrogen and/or sulfur compounds (4.6%) and carboxylic acids (0.8%). Among the identified volatiles, thiobismethane (commonly known as dimethylsulfide), methyl isocyanide and methyl isothiocyanate were found to be responsible for the nauseating and toxic smell emitted by the sponge and for the antimicrobial activity detected in the volatile extract. Exudation experiments in aquarium and in situ conditions revealed that thiobismethane, methyl isocyanide and methyl isothiocyanate are continuously released by the sponge. Upon injury, the concentration of these volatiles increased strongly. Hence, these substances form a chemical protective barrier which may help these sponges avoid fouling, compete for space, prevent infection in the short term, and/or signal generalist predators regarding the existence of other toxic substances in the internal tissues.     

The Influence of Oxygen on the Production of Volatile Compounds by Dipodascus aggregates

The production of volatile compounds by Dipodascus aggregatus was studied in relation to the oxygen concentration in the medium. Oxygen concentration was determined with a Clark oxygen electrode and volatile compounds in the atmosphere above the culture by a gas chromatographic technique. Shake cultures of the fungus in its stationary phase of growth were very sensitive to a decrease in oxygen concentration in the presence of residual glucose. Anaerobic conditions induced production of volatile compounds that continued for many hours. The pattern of production of volatile compounds observed under conditions of low oxygen concentration during the stationary phase of growth differed from that obtained under aerobic conditions during the exponential phase of growth.