Floral Aromatics of Ptelea: Chemical Identification and Human Response

Many members of the citrus family (Rutaceae) are valued for the aromatic compounds emitted by their flowers. Ptelea species are unusually cold-hardy members of the Rutaceae, but conflicting descriptions of the fragrance of their unisexual flowers may discourage the use of these trees. We analyzed floral volatiles and human response to these chemicals to test the hypothesis that the fragrance of staminate and pistillate flowers of these species differs. Gas chromatography and mass spectrometry showed that most volatile chemicals emitted by flowers of Ptelea trifoliata and Ptelea crenulata are monoterpenes, sesquiterpenes, and esters. Most volatiles were emitted from flowers of both sexes, but ethyl benzoate and estragole were emitted only from pistillate flowers. When concentrations of aromatics differed between sexes, they were higher for pistillate flowers, except for cis-3-hexenyl butanoate and an unidentified terpene. For P. crenulata and P. trifoliata, respectively, 81% and 77% of survey responses were from volunteers who liked the fragrance. Panelists most frequently described the scent of flowers of P. crenulata of both sexes with the words citrus, lime, and sweet. Panelists distinguished between pistillate and staminate flowers of P. trifoliata, describing the odor of pistillate flowers most frequently with the words damp-earthy, spicy, and sweet; staminate flowers were perceived as light, fresh, grassy, and pleasant. This work represents the first analysis of floral volatiles of P. crenulata and resolves conflicting prior reports regarding the floral fragrance of P. trifoliata. We conclude that differences among people rather than the sex of flowers account for conflicting prior reports of floral fragrance. The scents of flowers of P. crenulata and P. trifoliata appeal to most people and are horticultural assets of these trees.

Metabolite and Transcriptome Profiling Analysis Revealed That Melatonin Positively Regulates Floral Scent Production in Hedychium coronarium

Melatonin is a pleiotropic molecule that regulates a variety of developmental processes. Floral volatiles are important features of flowers that facilitate flower–visitor interactions by attracting pollinators, structure flower–visitor communities, and play defensive roles against plant and flower antagonists. Aside from their role in plants, floral volatiles are an essential ingredient in cosmetics, perfumes, pharmaceuticals, and flavorings. Herein, integrated metabolomic and transcriptomic approaches were carried out to analyze the changes triggered by melatonin exposure during the Hedychium coronarium flower development stages. Quantitative analysis of the volatiles of H. coronarium flowers revealed that volatile organic compound emission was significantly enhanced after melatonin exposure during the half bloom (HS), full bloom (FB) and fade stage (FS). Under the melatonin treatment, the emission of volatile contents was highest during the full bloom stage of the flower. Variable importance in projection (VIP) analysis and partial least-squares discriminant analysis (PLS-DA) identified 15 volatile compounds with VIP > 1 that were prominently altered by the melatonin treatments. According to the transcriptome sequencing data of the HS, FB, and FS of the flowers, 1,372, 1,510, and 1,488 differentially expressed genes were identified between CK-HS and 100MT-HS, CK-FB and 100MT-FB, and CK-FS and 100MT-FS, respectively. Among the significant differentially expressed genes (DEGs), 76 were significantly upregulated and directly involved in the floral scent biosynthesis process. In addition, certain volatile organic compounds were substantially linked with various DEGs after combining the metabolome and transcriptome datasets. Moreover, some transcription factors, such as MYB and bHLH, were also significantly upregulated in the comparison, which might be related to the floral aroma mechanism. Our results suggested that melatonin increased floral aroma production in H. coronarium flowers by modifying the expression level of genes involved in the floral scent biosynthesis pathway. These findings serve as a foundation for future research into the molecular mechanisms underlying the dynamic changes in volatile contents induced by melatonin treatment in H. coronarium.

Floral Volatiles: A Promising Method to Access the Rare Nocturnal and Crepuscular Bees

Crepuscular and/or nocturnal bees fly during the dusk, the dawn or part of the night. Due to their short foraging time and sampling bias toward diurnal bees, nocturnal bees are rarely collected and poorly studied. So far, they have been mostly sampled with light and Malaise traps. However, synthetic chemical compounds resembling floral volatiles were recently found to be a promising alternative to attract these bees. By reviewing available literature and collecting original data, we present information on the attraction and sampling of nocturnal bees with scent-baited traps. Bees were actively captured with entomological nets while approaching to filter papers moistened with distinct chemical compound, or passively caught in bottles with scent baits left during the night. So far, all data available are from the Neotropics. Nocturnal bees belonging to three genera, i.e., Ptiloglossa, Megalopta, and Megommation were attracted to at least ten different synthetic compounds and mixtures thereof, identified from bouquets of flowers with nocturnal anthesis. Aromatic compounds, such as 2-phenyletanol, eugenol and methyl salicylate, and the monoterpenoid eucalyptol were the most successful in attracting nocturnal bees. We highlight the effectiveness of olfactory methods to survey crepuscular and nocturnal bees using chemical compounds typically reported as floral scent constituents, and the possibility to record olfactory preferences of each bee species to specific compounds. We suggest to include this method in apifauna surveys in order to improve our current knowledge on the diversity of nocturnal bees in different ecosystems.

Generalized olfactory detection of floral volatiles in the highly specialized Greya-Lithophragma nursery pollination system

Volatiles are of key importance for host-plant recognition in insects. In the pollination system of Lithophragma flowers and Greya moths, moths are highly specialized on Lithophragma, in which they oviposit and thereby pollinate the flowers. Floral volatiles in Lithophragma are highly variable between species and populations, and moths prefer to oviposit into Lithophragma flowers from populations of the local host species. Here we used gas chromatography coupled with electroantennographic detection (GC-EAD) to test whether Greya moths detect specific key volatiles or respond broadly to many volatiles of Lithophragma flowers. We also addressed whether olfactory detection in Greya moths varies across populations, consistent with a co-evolutionary scenario. We analyzed flower volatile samples from three different species and five populations of Lithophragma occurring across a 1400 km range in the Western USA, and their sympatric female Greya politella moths. We showed that Greya politella detect a broad range of Lithophragma volatiles, with a total of 23 compounds being EAD active. We chemically identified 15 of these, including the chiral 6, 10, 14-trimethylpentadecan-2-one (hexahydrofarnesyl acetone), which was not previously detected in Lithophragma. All investigated Lithophragma species produced the (6R, 10R)-enantiomer of this compound. We showed that Greya moths detected not only volatiles of their local Lithophragma plants, but also those from allopatric populations/species that they not encounter in local populations. In conclusion, the generalized detection of volatiles and a lack of co-divergence between volatiles and olfactory detection may be of selective advantage for moths in tracking hosts with rapidly evolving, chemically diverse floral volatiles.

Comparative analysis of floral volatiles between the ‘Hass’ variety and Antillean race avocado

Mexico is the world's leading producer of ‘Hass’ avocado and the Antillean race avocado is grown in the south of the country. Avocado plant flowers produce a great variety of volatile compounds, which act as chemical signals to attract herbivores and pollinating insects, in addition to providing information for taxonomic purposes. The research aim was to identify and compare the floral volatiles between the ‘Hass’ and Antillean race avocado. Floral volatiles were captured by solid-phase microextraction, and identification was made by gas chromatography coupled to mass spectrometry (GC-MS). Thirty-five compounds were identified as constituents of the flower aromas; most were monoterpenes and sesquiterpenes. α-Pinene, sabinene, β-pinene, (E)-linalool oxide, benzyl nitrile, lavandulol, methyl salicylate, α-copaene, β-gurjunene and γ-muurolene were only found in ‘Hass’ avocado flowers. The differences can help classify the two types of avocados analyzed into different races. Eventually, this information could be used to find out if these volatile compounds influence the interactions of avocado with its pollinating insects and herbivores.

Headspace Volatiles of the Leaves and Flowers of Malvaviscus arboreus Cav. (Malvaceae)

Malvaviscus arboreus Cav., commonly known as Sleeping Hibiscus, is a plant species that belongs to the family Malvaceae with ornamental, culinary, and ethnomedical importance. This medicinal herb was reported to exhibit noteworthy antioxidant, cytotoxic, hepatoprotective, and anti-infective activities attributed to the presence of a variety of phytochemicals. In this work, the volatile compositions of the leaves and flowers of M. arboreus were studied and compared for the first time using the headspace gas chromatography-mass spectrometry (GC–MS) technique. Overall, 39 components were identified, comprising 36 from the leaves and 11 from the flowers, with an evident greater contribution of oxygenated compounds (89.54% in leaves and 89.35% in flowers) to their total volatiles. Phenolic ethers (41.64%) and ketones (21.57%) were the major chemical groups emitted by the flowers, while ketones (27.40%) and carboxylic acids (18.16%) dominated the volatile blends of the leaves. Anethole (32.32%), methyl isobutenyl ketone (19.18%), and methyl chavicol (9.32%) were the most abundant floral volatiles, whereas acetic acid (18.16%) was the major constituent given off by the leaves, followed by 2-cyclohexenone (9.60%) and anethole (7.39%). Additionally, from a biosynthetic point of view, the floral volatiles of M. arboreus showed the prevalence of phenylpropanoids/benzenoids (41.64%); however, fatty acid derivatives (54.30%) predominated among those produced by the leaves. The obtained results revealed noteworthy qualitative and quantitative variations in M. arboreus leaves and flowers' headspace volatiles that would help complement our phytochemical knowledge on this limitedly studied plant. Resumen. Malvaviscus arboreus Cav., comúnmente conocida como “Sleeping Hibiscus”, es una especie vegetal que pertenece a la familia Malvaceae con importancia ornamental, culinaria y etnomédica. Se ha reportado que esta hierba medicinal exhibe actividades antioxidantes, citotóxicas, hepatoprotectoras y antiinfecciosas notables que se atribuyen a la presencia de una variedad de fitoquímicos. En este trabajo se estudiaron las composiciones volátiles de las hojas y flores de M. arboreus y fueron comparadas utilizando la técnica de cromatografía de gases-espectrometría de masas (GC-MS). En total, se identificaron 39 componentes comprendiendo 36 de las hojas y 11 de las flores, con un evidente mayor aporte de compuestos oxigenados (89,54% en hojas y 89,35% en flores). Los éteres fenólicos (41,64%) y cetonas (21,57%) fueron los principales grupos químicos emitidos por las flores, mientras que las cetonas (27,40%) y los ácidos carboxílicos (18,16%) dominaron las mezclas volátiles de las hojas. Anetol (32,32%), metil isobutenil cetona (19,18%) y metil chavicol (9,32%) fueron los volátiles florales más abundantes, mientras que el ácido acético (18,16%) fue el principal componente desprendido por las hojas, seguido de la 2-ciclohexenona. (9,60%) y el anetol (7,39%). Adicionalmente, desde el punto de vista biosintético, los volátiles florales de M. arboreus mostraron la prevalencia de fenil propanoides/benzenoides (41,64%); sin embargo, los derivados de ácidos grasos (54,30%) predominaron entre los producidos por las hojas. Los resultados obtenidos revelaron variaciones cualitativas y cuantitativas notables en los volátiles de las hojas y flores de M. arboreus que ayudarán a complementar nuestro conocimiento fitoquímico en esta planta estudiada hasta ahora de forma limitada.

Composition of Strawberry Floral Volatiles and their Effects on Behavior of Strawberry Blossom Weevil, Anthonomus rubi

The strawberry blossom weevil (SBW), Anthonomus rubi, is a major pest in strawberry fields throughout Europe. Traps baited with aggregation pheromone are used for pest monitoring. However, a more effective lure is needed. For a number of pests, it has been shown that the attractiveness of a pheromone can be enhanced by host plant volatiles. The goal of this study was to explore floral volatile blends of different strawberry species (Fragaria x ananassa and Fragaria vesca) to identify compounds that might be used to improve the attractiveness of existing lures for SBW. Floral emissions of F. x a. varieties Sonata, Beltran, Korona, and of F. vesca, were collected by both solid-phase microextraction (SPME) and dynamic headspace sampling on Tenax. Analysis by gas chromatography/mass spectrometry showed the floral volatiles of F. x ananassa. and F. vesca were dominated by aromatic compounds and terpenoids, with 4-methoxybenzaldehyde (p-anisaldehyde) and α-muurolene the major compounds produced by the two species, respectively. Multi-dimensional scaling analyses separated the blends of the two species and explained differences between F. vesca genotypes and, to some degree, variation between F. x ananassa varieties In two-choice behavioral tests, SBW preferred odors of flowering strawberry plants to those of non-flowering plants, but weevils did not discriminate between odors from F. x ananassa and F. vesca flowering plants. Adding blends of six synthetic flower volatiles to non-flowering plants of both species increased the preference of SBW for these over the plants alone. When added individually to non-flowering plants, none of the components increased the preference of SBW, indicating a synergistic effect. However, SBW responded to 1,4-dimethoxybenzene, a major component of volatiles from F. viridis, previously found to synergize the attractiveness of the SBW aggregation pheromone in field studies.

Behavioral Responses of Thrips hawaiiensis (Thysanoptera: Thripidae) to Volatile Compounds Identified from Gardenia jasminoides Ellis (Gentianales: Rubiaceae)

Thrips hawaiiensis is a common thrips pest of various plant flowers with host preference. Plant volatiles provide important information for host-searching in insects. We examined the behavioral responses of T. hawaiiensis adults to the floral volatiles of Gardenia jasminoides Ellis, Gerbera jamesonii Bolus, Paeonia lactiflora Pallas, and Rosa chinensis Jacq. in a Y-tube olfactometer. T. hawaiiensis adults showed significantly different preferences to these four-flower plants, with the ranking of G. jasminoides > G. jamesonii > P. lactiflora ≥ R. chinensis. Further, 29 components were identified in the volatile profiles of G. jasminoides, and (Z)-3-hexenyl tiglate (14.38 %), linalool (27.45 %), and (E3,E7)-4,8,12-trimethyltrideca-1,3,7,11-tetraene (24.67 %) were the most abundant. Six-arm olfactometer bioassays showed that T. hawaiiensis had significant positive responses to (Z)-3-hexenyl tiglate, linalool, and (E3,E7)-4,8,12-trimethyltrideca-1,3,7,11-tetraene tested at various concentrations, with the most attractive ones being 10−3 μL/μL, 10−2 μL/μL and 100 μg/μL for each compound, respectively. In pairing of these three compounds at their optimal concentrations, T. hawaiiensis showed the preference ranking of (Z)-3-hexenyl tiglate > linalool > (E3,E7)-4,8,12-trimethyltrideca-1,3,7,11-tetraene. Large numbers of T. hawaiiensis have been observed on G. jasminoides flowers in the field, which might be caused by the high attraction of this pest to G. jasminoides floral volatiles shown in the present study. Our findings shed light on the olfactory cues routing host plant searching behavior in T. hawaiiensis, providing important information on how T. hawaiiensis targets particular host plants. The high attractiveness of the main compounds (e.g., linalool, (E3,E7)-4,8,12-trimethyltrideca-1,3,7,11-tetraene, particular (Z)-3-hexenyl tiglate) identified from volatiles of G. jasminoides flowers may be exploited further to develop novel monitoring and control tools (e.g., lure and kill strategies) against this flower-inhabiting thrips pest.