De Novo Synthesis of Dihydro-ß-ionone through Metabolic Engineering and Bacterium-Yeast Coculture.

Dihydro-ß-ionone is a common type of ionone used in the flavor and fragrance industries because of its characteristic scent. The production of flavors in microbial cell factories offers a sustainable and environmentally friendly approach to accessing them, independent of extraction from natural sources. However, the native pathway of dihydro-ß-ionone remains unclear, hindering heterologous biosynthesis in microbial hosts. Herein, we devised a microbial platform for de novo syntheses of dihydro-ß-ionone from a simple carbon source with glycerol. The complete dihydro-ß-ionone pathway was reconstructed in Escherichia coli with multiple metabolic engineering strategies to generate a strain capable of producing 8 mg/L of dihydro-ß-ionone, although this was accompanied by a surplus precursor ß-ionone in culture. To overcome this issue, Saccharomyces cerevisiae was identified as having a conversion rate for transforming ß-ionone to dihydro-ß-ionone that was higher than that of E. coli via whole-cell catalysis. Consequently, the titer of dihydro-ß-ionone was increased using the E. coli-S. cerevisiae coculture to 27 mg/L. Our study offers an efficient platform for biobased dihydro-ß-ionone production and extends coculture engineering to overproducing target molecules in extended metabolic pathways.

Organ-specific volatiles from Sonoran desert Krameria flowers as potential signals for oil-collecting bees.

The evolution of flowers that offer oils as rewards and are pollinated by specialized bees represents a distinctive theme in plant-pollinator co-diversification. Some plants that offer acetylated glycerols as floral oils emit diacetin, a volatile by-product of oil metabolism, which is utilized by oil-collecting bees as an index signal for the presence of floral oil. However, floral oils in the genus Krameria (Krameriaceae) contain ß-acetoxy-substituted fatty acids instead of acetylated glycerols, making them unlikely to emit diacetin as an oil-bee attractant. We analyzed floral headspace composition from K. bicolor and K. erecta, native to the Sonoran Desert of southwestern North America, in search of alternative candidates for volatile index signals. Using solid-phase microextraction, combined with gas chromatography-mass spectrometry, we identified 26 and 45 floral volatiles, respectively, from whole flowers and dissected flower parts of these two Krameria species. As expected, diacetin was not detected. Instead, ß-ionone emerged as a strong candidate for an index signal, as it was uniquely present in dissected oil-producing floral tissues (elaiophores) of K. bicolor, as well as the larval cells and provisions from its oil-bee pollinator, Centris cockerelli. This finding suggests that the floral oil of K. bicolor is perfused with ß-ionone in its tissue of origin and retains the distinctive raspberry-like scent of this volatile after being harvested by C. cockerelli bees. In contrast, the elaiophores of K. erecta, which are not thought to be pollinated by C. cockerelli, produced a blend of anise-related oxygenated aromatics not found in the elaiophores of K. bicolor. Our findings suggest that ß-ionone has the potential to impact oil-foraging by C. cockerelli bees through several potential mechanisms, including larval imprinting on scented provisions or innate or learned preferences by foraging adults.

Transcription factor VvWRKY70 inhibits both norisoprenoid and flavonol biosynthesis in grape.

Norisoprenoids and flavonols are important secondary metabolites in grape berries (Vitis vinifera L.). The former is a class of ubiquitous flavor and fragrance compounds produced by the cleavage of carotenoids, and the latter, which is derived from the flavonoid metabolic pathway, has been proposed as a general quality marker for red grapes. However, the transcriptional regulatory mechanisms underlying norisoprenoid and flavonol production are still not fully understood. In this study, we characterized a transcription factor, VvWRKY70, as a repressor of both norisoprenoid and flavonol biosynthesis in grape berries, and its expression was downregulated by light and high-temperature treatment. Overexpressing VvWRKY70 in grape calli reduced norisoprenoid and flavonol production, particularly under light exposure or at high temperature, by repressing the expression of several related genes in the isoprenoid and flavonoid metabolic pathways. VvWRKY70 downregulated ß-CAROTENE HYDROXYLASE 2 (VvBCH2) and CHALCONE SYNTHASE 3 (VvCHS3) expression based on yeast 1-hybrid analysis combined with electrophoretic mobility shift assay and chromatin immunoprecipitation-quantitative PCR. We discuss the role of VvWRKY70 in the coordinated regulatory network of isoprenoid and flavonoid metabolism. These findings provide a theoretical basis to improve flavor, color, and other comprehensive qualities of fruit crops and their processing products.

Postharvest UV-B exposure drives changes in primary metabolism, phenolic concentration, and volatilome profile in berries of different grape (Vitis vinifera L.) varieties.

BACKGROUND: The ultraviolet-B (UV-B) radiation can alter grape metabolism during berry development, but little is known on the effect of postharvest UV-B exposure. In this study, we evaluated the effect of postharvest UV-B exposure on berry primary and secondary metabolites in four grapevine varieties (Aleatico, Moscato bianco, Sangiovese, and Vermentino) in order to evaluate the possibility to increase the grape quality and its nutraceutical properties. RESULTS: The treatment did not significantly affect the berry primary metabolism in terms of organic acids, carbohydrates, and amino acids profile, regardless of the variety. UV-B exposure reduced the total anthocyanin content, particularly the tri-substituted and di-substituted forms in Aleatico and Sangiovese, respectively. An overall negative effect of UV-B irradiation on the flavonols profile of Aleatico, Moscato bianco, and Vermentino berries was found, whereas it enhanced the quercetin, myricetin and kaempferol concentration in Sangiovese. The free fraction of berry volatile organic compounds increased in UV-B-treated Aleatico and Moscato bianco berries, especially C13 -norisoprenoids and volatile phenols, as well as key monoterpenes, such as the linalool derivatives. However, higher concentrations of glycosylated monoterpenes and C13 -norisoprenoids were measured in Sangiovese and Vermentino berries treated with UV-B. CONCLUSION: This study provides new insights on the effect of postharvest UV-B radiation on berry secondary metabolism, highlighting a different modulation between varieties and suggesting the potential use of this technique to increase some nutraceutical and quality characteristics of grape berry. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Engineering a fermenting yeast able to produce the fragrant ß-ionone apocarotenoid for enhanced aroma properties in wine.

Wine is composed of multitudinous flavour components and volatile organic compounds that provide this beverage with its attractive properties of taste and aroma. The perceived quality of a wine can be attributed to the absolute and relative concentrations of favourable aroma compounds; hence, increasing the detectable levels of an attractive aroma, such as ß-ionone with its violet and berry notes, can improve the organoleptic qualities of given wine styles. We here describe the generation of a new grape-must fermenting strain of Saccharomyces cerevisiae that is capable of releasing ß-ionone through the heterologous expression of both the enzyme carotenoid cleavage dioxygenase 1 (CCD1) and its substrate, ß-carotene. Haploid laboratory strains of S. cerevisiae were constructed with and without integrated carotenogenic genes and transformed with a plasmid containing the genes of CCD1. These strains were then mated with a sporulated diploid wine industry yeast, VIN13, and four resultant crosses-designated MQ01-MQ04-which were capable of fermenting the must to dryness were compared for their ability to release ß-ionone. Analyses of their fermentation products showed that the MQ01 strain produced a high level of ß-ionone and offers a fermenting hybrid yeast with the potential to enhance the organoleptic qualities of wine.

The beneficial effect of leaf removal during fruit set on physiological, biochemical, and qualitative indices and volatile organic compound profile of the Cypriot reference cultivar 'Xynisteri'.

BACKGROUND: 'Xynisteri' is the reference Cypriot white cultivar that, despite its significant societal and economic impact, is poorly characterized regarding its qualitative properties, while scarce information exists regarding its aroma profile. In the current study, the effect of leaf removal during fruit set (BBCH 71) on 6-year cordon-trained, spur-pruned grapevines was assessed and an array of physiological, biochemical, and qualitative indices were monitored during successive developmental stages (BBCH 75, BBCH 85, BBCH 87, and BBCH 89). Grapes were additionally monitored for the volatile organic compounds (VOCs) profile during the advanced on-vine developmental stages (BBCH 85-BBCH 89) with the employment of gas chromatography-mass spectrometry (GC-MS), Fourier-transform near infrared (FT-NIR) spectra and electronic nose (E-nose) techniques. RESULTS: Grape berries from the vines subjected to leaf removal were characterized by higher solid soluble sugars (SSC), titratable acidity (TA), tartaric acid, and ammonium nitrogen contents, while this was not the case for assimilable amino nitrogen (primary amino nitrogen). A total of 75 compounds were identified and quantified, including aliphatic alcohols, benzenic compounds, phenols, vanillins, monoterpenes, and C13 -norisoprenoids. Leaf removal led to enhanced amounts of glycosylated aroma compounds, mainly monoterpenes, and C13 -norisoprenoids. Chemometric analysis, used through FT-NIR and E-nose, showed that the aromatic patterns detected were well associated to the grape ripening trend and differences between leaf removal-treated and control grapes were detectable during fully ripe stage. CONCLUSION: Leaf removal at fruit set resulted in an overall induction of secondary metabolism, with special reference to glycosylated aroma compounds, namely monoterpenes and C13 -norisoprenoids. © 2022 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Mediating oxidative stress enhances α-ionone biosynthesis and strain robustness during process scaling up.

BACKGROUND: α-Ionone is highly valued in cosmetics and perfumery with a global usage of 100-1000 tons per year. Metabolic engineering by microbial fermentation offers a promising way to produce natural (R)-α-ionone in a cost-effective manner. Apart from optimizing the metabolic pathways, the approach is also highly dependent on generating a robust strain which retains productivity during the scale-up process. To our knowledge, no study has investigated strain robustness while increasing α-ionone yield. RESULTS: Built on our previous work, here, we further increased α-ionone yield to 11.4 mg/L/OD in 1 mL tubes by overexpressing the bottleneck dioxygenase CCD1 and re-engineering the pathway, which is > 65% enhancement as compared to our previously best strain. However, the yield decreased greatly to 2.4 mg/L/OD when tested in 10 mL flasks. Further investigation uncovered an unexpected inhibition that excessive overexpression of CCD1 was accompanied with increased hydrogen peroxide (H2O2) production. Excessive H2O2 broke down lycopene, the precursor to α-ionone, leading to the decrease in α-ionone production in flasks. This proved that expressing too much CCD1 can lead to reduced production of α-ionone, despite CCD1 being the rate-limiting enzyme. Overexpressing the alkyl hydroperoxide reductase (ahpC/F) partially solved this issue and improved α-ionone yield to 5.0 mg/L/OD in flasks by reducing oxidative stress from H2O2. The strain exhibited improved robustness and produced ~ 700 mg/L in 5L bioreactors, the highest titer reported in the literature. CONCLUSION: Our study provides an insight on the importance of mediating the oxidative stress to improve strain robustness and microbial production of α-ionone during scaling up. This new strategy may be inspiring to the biosynthesis of other high-value apocarotenoids such as retinol and crocin, in which oxygenases are also involved.

Plant volatile ligands for male-biased MmedOBP14 stimulate orientation behavior of the parasitoid wasp Microplitis mediator.

As an important class of chemosensory-associated proteins, odorant binding proteins (OBPs) play a key role in the perception of olfactory signals for insects. Parasitoid wasp Microplitis mediator relies on its sensitive olfactory system to locate host larvae of Noctuidae and Geometridae. In the present study, MmedOBP14, a male-biased OBP in M. mediator, was functionally investigated. In fluorescence competitive binding assays, the recombinant MmedOBP14 showed strong binding abilities to five plant volatiles: ß-ionone, 3,4-dimethylacetophenone, 4-ethylacetophenone, acetophenone and ocimene. Homology modeling and molecular docking results indicated that the binding sites of all five ligands were similar and concentrated in the binding pocket of MmedOBP14. Except acetophenone, the remaining four ligands at 1, 10 and 100 µg/µL caused strong antennal electrophysiological responses in adults M. mediator, and males showed more obvious EAG responses to most ligands than females. In behavioral trials, males were attracted by low concentrations of MmedOBP14 ligands, whereas high doses of ß-ionone and acetophenone had a repellent effect on males. Moreover, 1 µg/µL of 3,4-dimethylacetophenone showed the strongest attractiveness to female wasps. These findings suggest that MmedOBP14 may play a more important role in the perception of plant volatiles for male wasps to locate habitat, supplement nutrition and search partners.

Genetic architecture of berry aroma compounds in a QTL (quantitative trait loci) mapping population of interspecific hybrid grapes (Vitis labruscana × Vitis vinifera).

BACKGROUND: Although grapes accumulate diverse groups of volatile compounds, their genetic regulation in different cultivars remains unelucidated. Therefore, this study investigated the volatile composition in the berries of an interspecific hybrid population from a Vitis labruscana 'Campbell Early' (CE) × Vitis vinifera 'Muscat of Alexandria' (MA) cross to understand the relationship among volatile compounds and their genetic regulation. Then, a quantitative trait locus (QTL) analysis of its volatile compounds was conducted. RESULTS: While MA contained higher concentrations of monoterpenes and norisoprenoids, CE contained higher concentrations of C6 compounds, lactones and shikimic acid derivatives, including volatiles characteristic to American hybrids, i.e., methyl anthranilate, o-aminoacetophenone and mesifurane. Furthermore, a cluster analysis of volatile profiles in the hybrid population discovered ten coordinately modulated free and bound volatile clusters. QTL analysis identified a major QTL on linkage group (LG) 5 in the MA map for 14 monoterpene concentrations, consistent with a previously reported locus. Additionally, several QTLs detected in the CE map affected the concentrations of specific monoterpenes, such as linalool, citronellol and 1,8-cineol, modifying the monoterpene composition in the berries. As for the concentrations of five norisoprenoids, a major common QTL on LG2 was discovered first in this study. Several QTLs with minor effects were also discovered in various volatile groups, such as lactones, alcohols and shikimic acid derivatives. CONCLUSIONS: An overview of the profiles of aroma compounds and their underlying QTLs in a population of interspecific hybrid grapes in which muscat flavor compounds and many other aroma compounds were mixed variously were elucidated. Coordinate modulation of the volatile clusters in the hybrid population suggested an independent mechanism for controlling the volatiles of each group. Accordingly, specific QTLs with significant effects were observed for terpenoids, norisoprenoids and some volatiles highly contained in CE berries.

Effects of different cultivation conditions on the production of ß-cyclocitral and ß-ionone in Microcystis aeruginosa.

BACKGROUND: Cyanobacteria blooms have become a major environmental problem and concern because of secondary metabolites produced by cyanobacteria released into the water. Cyanobacteria produce volatile organic compounds (VOCs), such as the compounds ß-cyclocitral and ß-ionone, which comprise odors, off-flavors, defense compounds, as well as growth regulators. Therefore, the general objective of this work was to evaluate the VOCs produced by two strains of Microcystis aeruginosa, differing in their ability to produce microcystins (LTPNA 01-non-producing and LTPNA 08-toxin-producing). The analysis of VOC production was carried out in (1) normal culture conditions, (2) under different light intensities (LI), and (3) after the external application of ß-ionone in both cultures. RESULTS: The results showed that ß-cyclocitral and ß-ionone are produced in all growth phases of LTPNA 01 and LTPNA 08. Both strains were producers of ß-cyclocitral and ß-ionone in normal culture conditions. It was observed that the ß-cyclocitral concentration was higher than ß-ionone in all light intensities investigated in this study. Additionally, the strain LTPNA 01 produced more ß-cyclocitral than LTPNA 08 at almost all times and LIs analyzed. However, the strain LTPNA 08 produced more ß-ionone, mainly at the initial times. In addition, the experiment results with the external addition of ß-ionone in the cultures showed that the strain LTPNA 01 produced more ß-cyclocitral in control conditions than in treatment. Nonetheless, ß-ionone production was higher in treatment conditions in LTPNA 08, indicating that the addition of ß-ionone may favor the production of these compounds and inhibit the production of ß-cyclocitral. CONCLUSION: Our results showed that some abiotic factors, such as different light intensities and external application of ß-ionone, can be triggers that lead to the production of VOCs.

One-pot synthesis of dihydro-ß-ionone from carotenoids using carotenoid cleavage dioxygenase and enoate reductase.

Dihydro-ß-ionone is a characteristic aroma compound of Osmanthus fragrans and is widely applied in the flavor & fragrance industry. However, the main focus is on chemical synthesis due to the metabolic pathways of dihydro-ß-ionone is still unclear. Here, we explored the one-pot synthesis system for dihydro-ß-ionone production using carotenoid cleavage dioxygenase (CCD) and enoate reductase. After screening the CCD enzyme, PhCCD1 from the Petunia hybrid was identified as the suitable enzyme for the first step of dihydro-ß-ionone synthesis due to the high enzyme activity for carotenoid. The PhCCD1 was expressed in Escherichia coli and further characterized. The optimal activity of PhCCD1 was observed at pH 6.8 and 45 °C. The enzyme was stable over the pH range of 6.0-8.0 and had good thermal stability below 40 °C. Then, we optimized the coupled reaction conditions for dihydro-ß-ionone production by PhCCD1 and enoate reductase AaDBR1 from Artemisia annua. Furthermore, we introduced the NADPH regeneration system with a 1.5-fold enhancement for dihydro-ß-ionone production. Collectively, approximately 13.34 mg/L dihydro-ß-ionone was obtained by the one-pot biosystem with a corresponding molar conversion of 85.8%. For the first time, we successfully designed and constructed a new synthesis pathway for dihydro-ß-ionone production in vitro. The coupled catalysis reported herein illustrates the feasibility of producing dihydro-ß-ionone from carotenoids and guides further engineering in the food industry.

Cloning and Characterization of a Novel Carotenoid Cleavage Dioxygenase 1 from Helianthus annuus.

Natural ß-ionone, a high-value flavoring agent, has been widely applied in the food, cosmetics, and perfume industry. However, attempts to overproduce ß-ionone in microorganisms have been limited by the efficiency of carotenoid cleavage dioxygenases (CCDs), which catalyzes ß-carotene in the biosynthesis pathway. In order to obtain CCD genes responsible for the specific cleavage of carotenoids generating ß-ionone, a novel carotenoid cleavage dioxygenase 1 from Helianthus annuus was cloned and overexpressed in Escherichia coli BL21(DE3). The recombinant CCD was able to cleave a variety of carotenoids at the 9, 10 (9', 10') sites to produce C13 products in vitro, including ß-ionone, pseudoionone, 3-hydroxy-4-oxo-ß-ionone, 3-hydroxy-ß-ionone, and 3-hydroxy-α-ionone, which vary depending on the carotenoid substrates. In comparison with lycopene and zeaxanthin, HaCCD1 also showed the high specificity for ß-carotene to cleave the 9, 10 (9', 10') double bond to produce ß-ionone in E. coli accumulating carotenoids. Finally, the expression of HaCCD1 in E. coli was optimized, and biochemical characterizations were further clarified. The optimal activity of HaCCD1 was at pH 8.8 and 50 °C. The Vmax for ß-apo-8'-carotenal was 10.14 U/mg, while the Km was 0.32 mM. Collectively, our study provides a valuable enzyme for the synthesis of natural ß-ionone by biotransformation and synthetic biology platform.

Asymmetric Total Synthesis and Biosynthetic Implications of Perovskones, Hydrangenone, and Hydrangenone B.

Perovskones and hydrangenones are a family of structurally complex triterpenoids that were mainly isolated from the genus Salvia medicinal plants. These isoprenoids exhibit a broad range of biological activities, such as antitumor and antiplasmodial activities. Here, we report the collective total synthesis of perovskone, perovskones C, D, F, hydrangenone, and hydrangenone B. The key strategies in this work include the following: (1) an asymmetric photoenolization/Diels-Alder reaction was developed to construct a tricyclic ring bearing three contiguous quaternary centers, which was used to build the core icetexane skeleton; (2) a bioinspired Diels-Alder reaction of perovskatone D with trans-α-ocimene was applied to stereospecifically generate perovskones; (3) late-stage oxidations and ring forming steps were developed to synthesize perovskones and hydrangenones. Our synthetic work suggests that (1) perovskatone D may serve as the precursor of the biosynthesis of perovskones and (2) the formation of hydrangenone and hydrangenone B, containing a five-membered D ring, may involve an oxidative ring cleavage and ring regeneration process.

ß-Ionone Attenuates Dexamethasone-Induced Suppression of Collagen and Hyaluronic Acid Synthesis in Human Dermal Fibroblasts.

Stress is a major contributing factor of skin aging, which is clinically characterized by wrinkles, loss of elasticity, and dryness. In particular, glucocorticoids are generally considered key hormones for promoting stress-induced skin aging through binding to glucocorticoid receptors (GRs). In this work, we aimed to investigate whether ß-ionone (a compound occurring in various foods such as carrots and almonds) attenuates dexamethasone-induced suppression of collagen and hyaluronic acid synthesis in human dermal fibroblasts, and to explore the mechanisms involved. We found that ß-ionone promoted collagen production dose-dependently and increased mRNA expression levels, including collagen type I α 1 chain (COL1A1) and COL1A2 in dexamethasone-treated human dermal fibroblasts. It also raised hyaluronic acid synthase mRNA expression and hyaluronic acid levels. Notably, ß-ionone inhibited cortisol binding to GR, subsequent dexamethasone-induced GR signaling, and the expression of several GR target genes. Our results reveal the strong potential of ß-ionone for preventing stress-induced skin aging and suggest that its effects are related to the inhibition of GR signaling in human dermal fibroblasts.

Meaningful Words in Crowd Noise: Searching for Volatiles Relevant to Carpenter Bees among the Diverse Scent Blends of Bee Flowers.

Olfactory cues constitute one of the most important plant-pollinator communication channels. Specific chemical components can be associated with specific pollinator functional groups due to pollinator-mediated selection on flower volatile (FV) emission. Here, we used multivariate analyses of FV data to detect an association between FVs and the worldwide distributed pollinator group of the carpenter bees (Xylocopa spp.). We compiled FVs of 29 plant species: 9 pollinated by carpenter bees, 20 pollinated by other bee pollinator functional groups. We tested whether FV emission differed between these groups. To rule out any phylogenetic bias in our dataset, we tested FV emission for phylogenetic signal. Finally, using field assays, we tested the attractive function of two FVs found to be associated with carpenter bees. We found no significant multivariate difference between the two plant groups FVs. However, seven FVs (five apocarotenoid terpenoids, one long-chain alkane and one benzenoid) were significantly associated with carpenter bee pollination, thus being "predictor" compounds of pollination by this pollinator functional group. From those, ß-ionone and (E)-methyl cinnamate presented the highest indicator values and had their behavioural function assessed in field assays. Phylogenetic signal for FVs emission was weak, suggesting that their emission could result from pollinator-mediated selection. In field assays, the apocarotenoid ß-ionone attracted carpenter bees, but also bees from other functional groups. The benzenoid (E)-methyl cinnamate did not attract significant numbers of pollinators. Thus, ß-ionone functions as a non-specific bee attractant, while apocarotenoid FVs emerge as consistent indicators of pollination by large food-foraging bees among bee-pollinated flowers.

Combined Metabolite and Transcriptome Profiling Reveals the Norisoprenoid Responses in Grape Berries to Abscisic Acid and Synthetic Auxin.

The abscisic acid (ABA) increase and auxin decline are both indicators of ripening initiation in grape berry, and norisoprenoid accumulation also starts at around the onset of ripening. However, the relationship between ABA, auxin, and norisoprenoids remains largely unknown, especially at the transcriptome level. To investigate the transcriptional and posttranscriptional regulation of the ABA and synthetic auxin 1-naphthaleneacetic acid (NAA) on norisoprenoid production, we performed time-series GC-MS and RNA-seq analyses on Vitis vinifera L. cv. Cabernet Sauvignon grape berries from pre-veraison to ripening. Higher levels of free norisoprenoids were found in ABA-treated mature berries in two consecutive seasons, and both free and total norisoprenoids were significantly increased by NAA in one season. The expression pattern of known norisoprenoid-associated genes in all samples and the up-regulation of specific alternative splicing isoforms of VviDXS and VviCRTISO in NAA-treated berries were predicted to contribute to the norisoprenoid accumulation in ABA and NAA-treated berries. Combined weighted gene co-expression network analysis (WGCNA) and DNA affinity purification sequencing (DAP-seq) analysis suggested that VviGATA26, and the previously identified switch genes of myb RADIALIS (VIT_207s0005g02730) and MAD-box (VIT_213s0158g00100) could be potential regulators of norisoprenoid accumulation. The positive effects of ABA on free norisoprenoids and NAA on total norisoprenoid accumulation were revealed in the commercially ripening berries. Since the endogenous ABA and auxin are sensitive to environmental factors, this finding provides new insights to develop viticultural practices for managing norisoprenoids in vineyards in response to changing climates.

Ionone Is More than a Violet's Fragrance: A Review.

The term ionone is derived from "iona" (Greek for violet) which refers to the violet scent and "ketone" due to its structure. Ionones can either be chemically synthesized or endogenously produced via asymmetric cleavage of ß-carotene by ß-carotene oxygenase 2 (BCO2). We recently proposed a possible metabolic pathway for the conversion of α-and ß-pinene into α-and ß-ionone. The differences between BCO1 and BCO2 suggest a unique physiological role of BCO2; implying that ß-ionone (one of BCO2 products) is involved in a prospective biological function. This review focuses on the effects of ionones and the postulated mechanisms or signaling cascades involved mediating these effects. ß-Ionone, whether of an endogenous or exogenous origin possesses a range of pharmacological effects including anticancer, chemopreventive, cancer promoting, melanogenesis, anti-inflammatory and antimicrobial actions. ß-Ionone mediates these effects via activation of olfactory receptor (OR51E2) and regulation of the activity or expression of cell cycle regulatory proteins, pro-apoptotic and anti-apoptotic proteins, HMG-CoA reductase and pro-inflammatory mediators. α-Ionone and ß-ionone derivatives exhibit anti-inflammatory, antimicrobial and anticancer effects, however the corresponding structure activity relationships are still inconclusive. Overall, data demonstrates that ionone is a promising scaffold for cancer, inflammation and infectious disease research and thus is more than simply a violet's fragrance.

UPLC-QTof-MSE Metabolomics Reveals Changes in Leaf Primary and Secondary Metabolism of Hop (Humulus lupulus L.) Plants under Drought Stress.

The hop (Humulus lupulus L.) is an important specialty crop used in beer production. Untargeted UPLC-QTof-MSE metabolomics was used to determine metabolite changes in the leaves of hop plants under varying degrees of drought stress. Principal component analysis revealed that drought treatments produced qualitatively distinct changes in the overall chemical composition of three out of four genotypes tested (i.e., Cascade, Sultana, and a wild var. neomexicanus accession but not Aurora), although differences among treatments were smaller than differences among genotypes. A total of 14 compounds consistently increased or decreased in response to drought stress, and this effect was generally progressive as the severity of drought increased. A total of 10 of these marker compounds were tentatively identified as follows: five glycerolipids, glutaric acid, pheophorbide A, abscisic acid, roseoside, and dihydromyricetin. Some of the observed metabolite changes likely occur across all plants under drought conditions, while others may be specific to hops or to the type of drought treatments performed.

Dehydration-Induced Carotenoid Cleavage Dioxygenase 1 Reveals a Novel Route for ß-Ionone Formation during Tea (Camellia sinensis) Withering.

ß-Ionone is a carotenoid-derived flavor and fragrance compound with a complex fruity and woody scent, known for its violet aroma. Due to the low odor threshold, ß-ionone dramatically affects the aroma and quality of tea. Previous studies have shown that ß-ionone increases during tea withering; however, its formation and regulation during the withering process are far from being understood. As dehydration is the most important stress during the withering of the tea leaves, we isolated a dehydration-induced gene belonging to the subfamily of carotenoid cleavage dioxygenases called carotenoid cleavage dioxygenase 1a (CsCCD1a) from Camellia sinensis and expressed it in Escherichia coli. The recombinant protein could generate volatile ß-ionone and pseudoionone from carotenoids. CsCCD1a was induced by dehydration stress, and its expression was related to the ß-ionone accumulation during tea withering. Overall, this study elucidated that CsCCD1a catalyzes the formation of ß-ionone in C. sinensis and enhanced the understanding of the ß-ionone formation under multiple stresses during the processing of tea.

Increased glycosidase activities improved the production of wine varietal odorants in mixed fermentation of P. fermentans and high antagonistic S. cerevisiae.

A selected Pichia fermentans strain was simultaneously and sequentially inoculated in synthetic and real juice with S. cerevisiae strains of different antagonistic activities in a ratio 1:1 to observe the correlation between varietal odorants and glycosidase activities. Fermentations using pure S. cerevisiae strains were used for comparison. Yeast biomass and glycosidase activities were monitored, varietal odorants were detected using HS-SPME-GC/MS during fermentation. The final wine aroma attributes were analyzed by trained panelists. Results showed that co-inoculation with high antagonistic S. cerevisiae resulted in higher glycosidase activities than others. Pearson correlation analysis indicated that yeast biomass was positively related to glycosidase activities during fermentation. The increase in glycosidase activities was the main reason for the higher production of terpenes and C13-norisoprenoids, and for the lower C6 compound content, which lead to superior fruity and floral aromas in the final wine samples of the high antagonistic S. cerevisiae group.