Scent Production Research Articles

Vanillin - Bioconversion and Bioengineering of the most popular plant flavour and its de novo biosynthesis in the vanilla orchid.

During recent years, biotechnology derived production of flavours and fragrances have expanded rapidly. The world's most popular flavour vanillin is no exception. This review outlines the current state of biotechnology-based vanillin synthesis with the use of ferulic acid, eugenol and glucose as substrates and bacteria, fungi and yeasts as microbial production hosts. The elucidated de novo biosynthetic pathway of vanillin in the vanilla orchid and the possible applied uses of this new knowledge in the biotechnology derived and pod-based vanillin industries are also highlighted.

Multi-capillary column-ion mobility spectrometry of volatile metabolites emitted by Saccharomyces cerevisiae.

Volatile organic compounds (VOCs) produced during microbial fermentations determine the flavor of fermented food and are of interest for the production of fragrances or food additives. However, the microbial synthesis of these compounds from simple carbon sources has not been well investigated so far. Here, we analyzed the headspace over glucose minimal salt medium cultures of Saccharomyces cerevisiae using multi-capillary column-ion mobility spectrometry (MCC-IMS). The high sensitivity and fast data acquisition of the MCC-IMS enabled online analysis of the fermentation off-gas and 19 specific signals were determined. To four of these volatile compounds, we could assign the metabolites ethanol, 2-pentanone, isobutyric acid, and 2,3-hexanedione by MCC-IMS measurements of pure standards and cross validation with thermal desorption–gas chromatography-mass spectrometry measurements. Despite the huge biochemical knowledge of the biochemistry of the model organism S. cerevisiae, only the biosynthetic pathways for ethanol and isobutyric acid are fully understood, demonstrating the considerable lack of research of volatile metabolites. As monitoring of VOCs produced during microbial fermentations can give valuable insight into the metabolic state of the organism, fast and non-invasive MCC-IMS analyses provide valuable data for process control.

Floral scent contributes to interaction specificity in coevolving plants and their insect pollinators.

Chemical defenses, repellents, and attractants are important shapers of species interactions. Chemical attractants could contribute to the divergence of coevolving plant-insect interactions, if pollinators are especially responsive to signals from the local plant species. We experimentally investigated patterns of daily floral scent production in three Lithophragma species (Saxifragaceae) that are geographically isolated and tested how scent divergence affects attraction of their major pollinator-the floral parasitic moth Greya politella (Prodoxidae). These moths oviposit through the corolla while simultaneously pollinating the flower with pollen adhering to the abdomen. The complex and species-specific floral scent profiles were emitted in higher amounts during the day, when these day-flying moths are active. There was minimal divergence found in petal color, which is another potential floral attractant. Female moths responded most strongly to scent from their local host species in olfactometer bioassays, and were more likely to oviposit in, and thereby pollinate, their local host species in no-choice trials. The results suggest that floral scent is an important attractant in this interaction. Local specialization in the pollinator response to a highly specific plant chemistry, thus, has the potential to contribute importantly to patterns of interaction specificity among coevolving plants and highly specialized pollinators.

Effects of salicylic acid on quality and quantity of essential oil components in Lippia citriodora H.B.K.

The world production and consumption of essential oils and perfumes are increasing very fast. Exogenous salicylic acid (SA) was effective in inducing secondary metabolites formation in plant cell culture or in vivo plants. To study the effect of exogenous application of salicylic acid on volatile oil content and composition of lemon verbena (Lippia citriodora H.B.K.), an experiment was conducted as a Randomized Complete Block Design (RCBD) with three replicates. Experimental treatments included salicylic acid at four levels (2.5, 5, 10 and 20 mM), distillated water and control (non treat). It’s essential oil was analyzed by GC and GC/MS. Results showed that SA application increased content of β-Pinene, 1,8-Cineol, ɣ-Terpinene, Terpinolene, cis limonene oxide, α-terpineol, neral, α-terpinyl acetate, geranyl acetate, ɣ-elemene, α-humulene, cubenol, spathulenol, globulol and epi-α-cadinol. Some compounds such as α-Pinene, sabinene, limonene and citronellal were reduced by SA treatments. The yield of essential oil was highest at 10 mM (0.71%). The findings suggest that SA at 10 mM was the best treatment considering the measured compounds.

Classification of perfume products using gas chromatography-mass spectrometry and pattern recognition

Perfume is a mixture of fragrant essential oils and aroma compounds, fixatives, and solvents used to give human body, objects, and living spaces a pleasant odour. Perfume compositions are mixture of natural and synthetic sources which consist of 100 or more components. The difference in composition of perfume is the main characteristics which can classify the perfume into several groups such as animal and plant sources. Therefore, correct identification of each perfume product composition is crucial in order to do the classification. The perfume products have been analyzed using gas chromatography mass spectrometry and chemometric technique. There are 34 samples of perfume products used in this article which is taken from different brand available around Skudai. Chemometric technique was applied in this research to simplify data obtained by allowing them to form classes to their respective groups. Two unsupervised pattern recognition technique namely principal component analysis (PCA) and cluster analysis were used. The perfume product samples investigated in this research can be classified into two main groups namely animal and plant sources. Principal component analysis (PCA) and cluster analysis were successfully used in classifying the perfume product according to their respective groups based on the compositional information from the chromatogram of each samples.________________________________________GRAPHICAL ABSTRACT

Perfume production in flowers of Angelonia salicariifolia attracts males of Euglossa annectans which do not promote pollination

Male Euglossini bees (Apidae) forage for floral and non-floral perfumes. Although the use of these perfumes has not yet been clearly reported, there are indications of using perfume from varied exogenous sources as a pheromone analogue since male orchid bees do not produce it. Here, we report an interaction between male Euglossa annectans and flowers of Angelonia salicariifolia (Plantaginaceae). Floral anatomy analysis and a histological study of two different sets of trichomes indicate the presence of aromatic oils (perfume) in one set of these trichomes. The other set of trichomes produce resins. Both sets of trichomes are found in the same floral area of the flower where the male orchid bees were seen. The male orchid bees studied forage for perfumes but do not promote pollination since they do not reach the reproductive structures during their visit. Our records show that only male E. annectans were attracted to the volatile fragrance although we know that other orchid bee species also occur in the area.

Application of Taiwan aboriginal myths to cultural perfume products

Application of Taiwan aboriginal myths to cultural perfume products

Is the timing of scent emission correlated with insect visitor activity and pollination in long-spurred Satyrium species?

Plants are expected to emit floral scent when their pollinators are most active. In the case of long-tubed flowers specialised for pollination by crepuscular or nocturnal moths, scent emissions would be expected to peak during dawn. Although this classic idea has existed for decades, it has rarely been tested quantitatively. We investigated the timing of flower visitation, pollination and floral scent emissions in six long-spurred Satyrium species (Orchidaceae). We observed multiple evening visits by pollinaria-bearing moths on flowers of all study species, but rarely any diurnal visits. The assemblages of moth pollinators differed among Satyrium species, even those that co-flowered, and the lengths of moth tongues and floral nectar spurs were strongly correlated, suggesting that the available moth pollinator fauna is partitioned by floral traits. Pollinarium removal occurred more frequently during the night than during the day in four of the six species. Scent emission, however, was only significantly higher at dusk than midday in two species. Analysis of floral volatiles using gas chromatography coupled with mass spectrometry yielded 168 scent compounds, of which 112 were species-specific. The scent blends emitted by each species occupy discrete clusters in two-dimensional phenotype space, based on multivariate analysis. We conclude that these long-spurred Satyrium species are ecologically specialised for moth pollination, yet the timing of their scent emission is not closely correlated with moth pollination activity. Scent composition was also more variable than expected from a group of closely related plants sharing the same pollinator functional group. These findings reveal a need for greater understanding of mechanisms of scent production and their constraints, as well as the underlying reasons for divergent scent chemistry among closely related plants.

Biotechnology of Scented Roses: A Review

Scented rose species are very important in the genus Rosa due to their use in highly-prized essential oil production and as garden roses. The fragrance of the rose flower (attar) captured by extraction is one of the most valuable flavor and fragrance products. A comprehensive study of the papers published in the field of in vitro culture and biotechnology of these species, particularly Damask rose, was performed. Then, in separate sections, the articles related to media preparation for callus culture, shoot regeneration and multiplication, rooting, and cell suspension culture were reviewed. In this paper, the stages of the in vitro culture of scented roses, including establishment, multiplication, in vitro rooting, acclimatization, and genetic variability are also considered. Subsequently, investigations on methods of in vitro culture of scented roses by callus, pollen, anther, cell suspension, and protoplast culture are described. Finally, technological improvements for micropropagation, biotechnology. pharmaceutical, and other properties of the Damask rose and other species of scented roses are discussed. Prospects of in vitro culture and biotechnological techniques of scented rose species are also discussed. Among scented roses, the focus of this paper is on four species, namely Rosa damascena Mill., R. gallica Linn., R. centifolia Linn., and R. moschata Herrm., which are highly prized for oil production.

The male gender bias in autism may be due to preferential depletions of oxytocin and arginine‐vasopressin receptors positive neurons exposed to certain fragrances during fetal development (LB502)

The male gender bias in autism may be due to preferential depletions of oxytocin and arginine‐vasopressin receptors positive neurons exposed to certain fragrances during fetal development Leanna Sealey and Omar Bagasra M.D. Ph.D.Autism spectrum disorders (ASDs) are developmental conditions characterized by deficits in social interaction, verbal and nonverbal communication, and obsessive/stereotyped patterns of behavior. There is also evidence of impoverished language and empathy, and a profound inability to adopt another's viewpoint ‐ a failure to construct a “theory of mind” for interpreting another person's thoughts and intentions. In developed countries, it is now reported that 1%–1.5% of children have ASD, and in the US 2013 CDC report, approximately one in 80 children suffer from ASD. Although there is no reliable neurophysiological marker associated with ASDs, low levels of plasma oxytocin (OXY) and arginine‐vasopressin (AVP) have been reported. We have hypothesized that synthetic fragrances, which many of us are exposed to on a daily have potential to cause neurological damage to developing fetuses.Notably, the alarming rise in ASD parallels the rise in production of synthetic fragrances from petrochemicals and carcinogenic benzene‐based compounds. Numerous studies have shown that children with autism have significantly lower levels of OXY in plasma samples than their typical peers. Furthermore, in normal children, lower concentrations of OXY in plasma are associated with lower social and cognitive functioning. Here we show that male neuroblastoma cell lines, but not the female, exposure to fentomoles concentrations of certain fragments results in depletions of both OXY and AVP receptor + neurons. Our studies are the first to reveal a direct connection between fragrance exposures as the potential neuropathic agent that may cause autism and explains the gender bias.Grant Funding Source: Department of Education

Short-Chain Flavor Ester Synthesis in Organic Media by an E. coli Whole-Cell Biocatalyst Expressing a Newly Characterized Heterologous Lipase

Short-chain aliphatic esters are small volatile molecules that produce fruity and pleasant aromas and flavors. Most of these esters are artificially produced or extracted from natural sources at high cost. It is, however, possible to ‘naturally’ produce these molecules using biocatalysts such as lipases and esterases. A gene coding for a newly uncovered lipase was isolated from a previous metagenomic study and cloned into E. coli BL21 (DE3) for overexpression using the pET16b plasmid. Using this recombinant strain as a whole-cell biocatalyst, short chain esters were efficiently synthesized by transesterification and esterification reactions in organic media. The recombinant lipase (LipIAF5-2) showed good affinity toward glyceryl trioctanoate and the highest conversion yields were obtained for the transesterification of glyceryl triacetate with methanol. Using a simple cetyl-trimethylammonium bromide pretreatment increased the synthetic activity by a six-fold factor and the whole-cell biocatalyst showed the highest activity at 40°C with a relatively high water content of 10% (w/w). The whole-cell biocatalyst showed excellent tolerance to alcohol and short-chain fatty acid denaturation. Substrate affinity was equally effective with all primary alcohols tested as acyl acceptors, with a slight preference for methanol. The best transesterification conversion of 50 mmol glyceryl triacetate into isoamyl acetate (banana fragrance) provided near 100% yield after 24 hours using 10% biocatalyst loading (w/w) in a fluidized bed reactor, allowing recycling of the biocatalyst up to five times. These results show promising potential for an industrial approach aimed at the biosynthesis of short-chain esters, namely for natural flavor and fragrance production in micro-aqueous media.

Floral fraudulence: Do blue Thelymitra species (Orchidaceae) mimic Orthrosanthus laxus (Iridaceae)?

In Western Australia, Thelymitra crinita Lindl. and T. macrophylla Lindl. are pollinated by female, polylectic bees but offer no edible rewards. Flowers of Orthrosanthus laxus (Endl.) Benth. (Iridaceae) offer granular pollen and previous authorities suggest it is a Batesian model of T. crinita. We analyzed the floral fragrances and measured the floral dimensions of the orchid species, their putative hybrid, and O. laxus. Although the ‘scentless’ T. crinita emitted low levels of monoterpenoids and sesquiterpenoids, the pleasantly discernible fragrance of T. macrophylla was dominated by 2-phenylethanol. Their putative hybrid produced slightly lower levels of 2-phenylethanol compared with T. macrophylla and failed to produce any sesquiterpenoids associated with T. crinita. However, the hybrid produced higher volumes of the monoterpene linalool than either parent species. The fragrance of O. laxus contained 2-phenylethanol but lacked the sesquiterpenoids. We also measured perianth area and symmetry as well as the length and width of contrastingly pigmented floral centres for each taxon. Significant differences in floral area and symmetry were detected between the putative hybrid, the two parent species, and O. laxus. In contrast, the floral reward centre area (tuft of stamens) in O. laxus was significantly larger than the pseudo-reward centres (mitras) of both Thelymitra species and their hybrid. At the peak of their respective, but overlapping flowering periods, an inflorescence of T. macrophylla produced more than twice the number of open flowers as T. crinita and more than four times the number of open flowers on cymes of O. laxus. Based on scent production and visual displays, T. macrophylla appears more likely to be a Batesian floral mimic of O. laxus. We suggest that large-flowered Thelymitra species appear to produce a novel, visual and olfactory attractant pattern of fraudulence we call the ‘New Again, More Again Effect’.

Arabidopsis thaliana for testing the phytotoxicity of volatile organic compounds

Geosmin and C-8 hydrocarbons are among the major volatile organic compounds (VOC) responsible for the distinctive, musty odor of filamentous fungi. In this study, we developed a plant bioassay for testing the possible toxicity of these compounds, as well as four air freshener products sometimes used to mask their odor. Seeds and vegetative plants of Arabidopsis thaliana were exposed to 1 ppm of 14 different volatile treatments (both single compounds and mixtures) for 72 h and monitored for germination rate, seedling formation, vegetative plant vigor and chlorophyll concentration. All VOCs tested had some inhibitory effect on seed germination or seedling formation; 1-octen-3-one was the most active, giving almost complete inhibition of germination. Geosmin did not prevent germination (radicle protrusion) but seedling formation was arrested 90 %. Of solvents and fragrance products tested, only the scented oil product was as active as the C-8 biogenic compounds in inhibiting seed germination and seedling formation. Two-week-old plants exposed to 1 ppm of individual fungal VOCs for 72 h all exhibited some degree of stress symptoms including smaller leaf size and weight, discoloration, leaf curling, small necrotic lesions, and reduced chlorophyll concentration. Two-week-old vegetative plants exposed to solvents and air freshener products were generally smaller in size. The single most phytotoxic compound tested was 1-octen-3-one which almost completely inhibited seed germination and was lethal to vegetative phase plants. Formaldehyde at 1 ppm killed 2-week-old seedlings but had little effect on seed germination. In conclusion, the A. thaliana bioassay provides an inexpensive approach for testing the toxicity of gas phase molecules.

Optimal process conditions for zeolite catalyzed acylation of anisole

Acylation of anisole is a commercially important reaction in the production of various fine chemicals, agrochemicals, pharmaceuticals and fragrances. Conventionally, it is carried out using the catalysts like AlCl3, FeCl3, ZnCl3, HF, which suffer from major drawbacks such as larger consumption, corrosion and safety issues, waste disposal and the material handling. Hence the conventional catalysts are being replaced with solid acid catalysts like H-Beta, H-ZSM5 to overcome the above drawbacks. In this work, liquid phase acylation of anisole has been carried out employing H Beta, H ZSM-5 and HY catalysts and the process standardization at a macrolevel has been done with reference to parameters like temperature, reaction time, molar ratio of reactants, catalyst nature, Si/Al ratio of catalyst and the catalyst quantity. In addition, catalyst stability was investigated.

Ecological relationship between floral thermogenesis and pollination in Nelumbo lutea (Nelumbonaceae)

Floral thermogenesis is an unusual floral trait with a well-documented physiological process, and yet, there is limited understanding of how this trait influences plant reproduction. The current study was undertaken to gain a better understanding of how floral thermogenesis in Nelumbo lutea impacts pollinator attraction and consequent plant reproduction. We conducted field studies on floral thermogenesis and thermoregulation, flower sexual development, floral visitation patterns, breeding system, pollen transfer dynamics, and floral scent production. The most abundant visitors to the thermoregulatory flowers included the Phoridae (Diptera), Chrysomelidae (Coleoptera), and Hymenoptera. Chrysomelid beetles, particularly Diabrotica, were frequent visitors to both first-day female- and second-day bisexual-phase flowers, while phorid flies were most common in bisexual-phase flowers. Pollen transfer experiments indicated that Diabrotica was equally effective in depositing pollen on stigmas, as were the less frequent, but pollen-loaded halictid bees. Flowers received a taxonomically wide assemblage of floral visitors and appear adapted to attract beetles, primarily Chrysomelidae and medium-sized bees. This study is the first to provide strong support that beetles can comprise the dominant portion of floral visitors and are as effective in pollen transfer as bees. Thermogenesis aids in dispersing the main floral scent component-1,4-dimethoxybenzene-attracting both chrysomelids and bees, while thermoregulation causes chrysomelid beetles to actively seek out new flowers for evening residence. This search behavior likely results in chrysomelids affecting cross-pollination.

Bioinformatics Analysis of Phenylacetaldehyde Synthase (PAAS), a Protein Involved in Flower Scent Production, in Rose

Rose flowers produce and emit the aromatic volatiles 2-phenylacetaldehyde (PAA) and 2-phenylethanol (2-PE), which have a distinctive flowery/rose-like scent. Previous studies of rose have shown that, similar to Petunia flowers, PAA is formed from L-phenylalanine via pyridoxal-5'-phosphate-dependent L-aromatic amino acid decarboxylase. Rosa phenylacetaldehyde synthase sequence (RhPAAS) is homologous to Petunia phenylacetaldehyde synthase (PhPAAS). Since there is not much experimental data available about different structural properties of that PAAS protein, in the present investigation, we studied the different structural properties of the PAAS protein in petunia and rose using bioinformatics tools. The features of the first, secondary and tertiary structures of this protein were compared between Petunia and Rose. The results indicated that the frequency of negatively charged, Leucine, and frequency of the Ser-Leu, Pro-Glu , Phe-Ser and the, Thr-Thr dipeptides in petunia are more than those in Rose. In contrast, in petunia, the frequencies of hydrophobic and hydrophilic residues, α-helix, β-sheet, β-strands of petunia are lower than those in rose. The features achieved in this study may also provide useful clues for designing scent production pathways using protein engineering techniques.

Spermatotoxicity in Animal Models Exposed to Fragrance Components

Various commonly-used products have been reported to contain chemicals that could disrupt estrogen and testosterone hormone. As trade secrets, these chemicals are generally listed as fragrance to mask individual identity. In this study, the reproductive implications of two commonly used perfumes (designated as F1 and F2) in Nigeria were carried out. Sixty adult male wistar rats (12-13 weeks old) were randomly divided into four groups (A-F) of ten rats each. Group A and B rats were exposed to 5 mL kg-1 b.wt. of normal saline for 56 days and 112 days, respectively via whole body inhalation. Group C and D rats were exposed to 5 mL kg-1 b.wt. of a fragrance product designated as F1 for a period of 56 days and 112 days, respectively while Group E and Group F rats were exposed to 5 mL kg-1 b.wt. of a designated fragrance product F2 for a period of 56 days and 112 days, respectively. The results obtained from this study showed a significant (p 0.005) when compared to values of the control group A and B. Based on the spermiographic evaluation from this study, fragrance materials could have an adverse effect on spermatozoa of the intact male wistar rats.

Influence of N- and/or C-terminal regions on activity, expression, characteristics and structure of lipase from Geobacillus sp. 95

GD-95 lipase from Geobacillus sp. strain 95 and its modified variants lacking N-terminal signal peptide and/or 10 or 20 C-terminal amino acids were successfully cloned, expressed and purified. To our knowledge, GD-95 lipase precursor (Pre-GD-95) is the first Geobacillus lipase possessing more than 80% lipolytic activity at 5 °C. It has maximum activity at 55 °C and displays a broad pH activity range. GD-95 lipase was shown to hydrolyze p-NP dodecanoate, tricaprylin and canola oil better than other analyzed substrates. Structural and sequence alignments of bacterial lipases and GD-95 lipase revealed that the C-terminus forms an α helix, which is a conserved structure in lipases from Pseudomonas, Clostridium or Staphylococcus bacteria. This work demonstrates that 10 and 20 C-terminal amino acids of GD-95 lipase significantly affect stability and other physicochemical properties of this enzyme, which has never been reported before and can help create lipases with more specific properties for industrial application. GD-95 lipase and its modified variants GD-95-10 can be successfully applied to biofuel production, in leather and pulp industries, for the production of cosmetics or perfumes. These lipases are potential biocatalysts in processes, which require extreme conditions: low or high temperature, strongly acidic or alkaline environment and various organic solvents.

A Green Approach for Producing Solvent-free Anisyl Acetate by Enzymecatalyzed Direct Esterification in Sponge-like Ionic Liquids Under Conventional and Microwave Heating

The biocatalytic synthesis of anisyl acetate fragrance was carried out by direct esterification of acetic acid with anisyl alcohol in sponge-like ionic liquids (SLILs), e.g. N,N,N,N-hexadecyltrimethylammonium bis(trifluoromethylsulfonyl) imide ([C16tma][NTf2], N,N,N,N-octadecyltrimethylammonium bis(trifluoromethylsulfonyl)imide ([C18tma] [NTf2]), etc. as reaction/separation media under conventional and microwave (MW) heating. These SLILs are temperatureswitchable ionic liquid/solid phases that behave as sponges. As liquid phases, they are excellent monophasic reaction media for the lipase-catalyzed synthesis of anisyl acetate, the product yield being improved up to 100% for 2 hour reaction under the appropriate reaction conditions (i.e. SLIL concentration, alcohol: acid molar ratio, enzyme amount, dehydrating molecular sieves, temperature and MW heating). As a function of the phase behaviour of different SLIL/anisyl acetate mixtures, a new clean separation protocol based on the centrifugation of the solid IL/flavour ester through nylon membranes was proposed, which provided a nearly full separation of the solid SLIL and the easy recovery of the reaction mixture. The enzymatic synthesis of anisyl acetate in [C16tma] [NTf2] under MW assistance, followed by the separation step of the solid SLIL, provided a nearly solvent-free fragrance product with up to 0.89 g/mL concentration. The catalytic activity of the enzyme / SLIL system remained unchanged for ten consecutive operational cycles. This work reports a straightforward and sustainable approach for producing anisyl acetate as a natural flavour and demonstrates to be suitable for scaling-up, providing a high potential for practical application. Keywords: Anisyl acetate, anisyl alcohol, biocatalysis, biocatalytic process, clean process, enzymatic synthesis, esterification, flavours, fragrances, green chemistry, ionic liquids, lipase, neoteric solvents, pure products, solvent-free, sponge-like ionic liquids, sustainable chemistry, switchable ionic liquids.

Postharvest physiology and volatile production by flowers of Ptilotus nobilis

Ptilotus nobilis (Lindl.) F. Muell. has potential in the floriculture industries as a cut flower crop. Ethylene production and respiration rates, fresh weight changes and volatile scent production from cut inflorescences of P. nobilis cultivars Passion (dark pink flowers) and Purity (white-green flowers) were measured during vase life. Inflorescence weight loss was significant (P<0.001) during vase life with wilting and colour loss being the primary reasons for loss of vase life. Inflorescences ready for the cut market stored and at 22°C had vase lives of >12d. Ethylene production by inflorescences was low to negligible. Treatment with silverthiosulphate (STS) and ethylene had no effects on vase life. Evidently, ethylene did not play a role in determining the postharvest longevity of cut P. nobilis flowers. Respiration rates of inflorescences were high at harvest (>700mg CO2kg−1 FWh−1) and declined gradually thereafter during vase life. Total volatile emissions followed a similar pattern. For Passion, respiration rates of immature florets were significantly greater (P=0.02) than florets from other developmental stages while the calyx produced the most CO2. For Purity, respiration rates of florets of different maturities did not differ and the reproductive tissue produced the most CO2. Only fully opened mature florets with their stigma and anthers revealed, emitted significant quantities of volatiles (P<0.001) and primarily from the calyx tissue for both cultivars. The individual volatiles differed somewhat for the two cultivars. However, both produced significant quantities of benzaldehyde, 3,5-dimethoxytoluene and benzyl alcohol. These compounds have previously been associated with desirable floral scent.