Plant Volatile Compounds Research Articles

Olfactory attraction mediated by the maxillary palps in the striped fruit fly, Bactrocera scutellata: Electrophysiological and behavioral study

Here, we report that the olfactory attraction of the striped fruit fly, Bactrocera scutellata (Hendel; Diptera: Tephritidae), a serious pest of pumpkin and other cucurbitaceae plants, to cue lure and raspberry ketone is mediated by the maxillary palps. The antennae, bearing three morphological types (basiconic, trichoid, and coeloconic) of olfactory sensilla, in male and female B. scutellata exhibited significant electroantennogram (EAG) responses to a plant volatile compound, 3-octanone, and methyl eugenol, whereas cue lure, raspberry ketone, and zingerone that are known to attract several other species of Bactrocera fruit flies elicited no significant EAG responses from both sexes. In contrast, maxillary palps, housing one morphological type of basiconic sensilla, displayed the largest electropalpogram (EPG) responses to cue lure followed by raspberry ketone among the five compounds tested in male and female B. scutellata, with only minor EPG responses to 3-octanone, which indicates that the maxillary palps are responsible for detecting cue lure and raspberry ketone in this species. In field trapping experiments, significant number of male B. scutellata were captured in the traps baited with cue lure or raspberry ketone, in which the attractiveness of cue lure was significantly higher than that of raspberry ketone. Methyl eugenol and zingerone were not behaviorally attractive to B. scutellata although they elicited significant EPG responses. Our study indicates that the behavioral attraction of B. scutellata to cue lure and raspberry ketone is mediated by the olfactory sensory neurons present in the maxillary palps.

Perspective from grad school: Finding motivation amid challenges in Nigeria

As a graduate student in a developing nation like Nigeria, I face lots of challenges. My research deals with extraction, characterization, and biological activities of plant volatile compounds from aromatic plants grown in Nigeria. Hindered by an unstable power supply, I have to monitor the stability of electricity to carry out my experiments. This has extended my course of study. I also don’t have access to necessary analytical instruments like ultraviolet spectrophotometers and infrared, mass, and nuclear magnetic resonance spectrometers. I overcame these challenges by collaborating with research institutes such as Nigeria’s National Research Institute for Chemical Technology, where such resources are available. There is no research body or foundation in place that provides grants, scholarships, or loans to facilitate the successful and timely completion of my program. This has required me to spend huge amounts of money on tuition, accommodations, and travel to both local and international conferences. For

Quality and intensity of light affect Lippia gracilis Schauer plant growth and volatile compounds in vitro

The aim of this study was to evaluate the effects of different intensities and quality of light and explant type on the growth of and volatile compounds in Lippia gracilis in vitro. The treatments were as follows: light intensities of 26, 51, 69, 94, or 130 µmol m−2 s−1 from fluorescent lamps and light-emitting diode (LED) lamps at different wavelengths, namely, white, red, blue, and combinations of red and blue light at ratios of 2.5:1 and 1:2.5, respectively, and two explant types, namely, nodal and apical segments. On the 30th day of culture on half-strength Murashige and Skoog (Physiol Plant 15(3):473–497, 1962) medium, growth, production of photosynthetic pigments, chlorophyll a and b, total chlorophyll, carotenoids, and volatile constituents (using headspace gas chromatography-mass spectrometry) were analyzed. The light quality and intensity significantly influenced the in vitro growth of L. gracilis. The apical segments were superior in all parameters evaluated compared to nodal segments. The number of segments plantlet−1, root length, and leaf, shoot, root, and total weight were higher with increasing light intensity, especially under the 94 µmol m−2 s−1 treatment, for both explant types. The red light showed the highest leaf (32.28 mg plantlet−1) and total (58.33 mg plantlet−1) dry weight of all the light qualities. Major constituents, namely, ρ-cymene, γ-terpinene, thymol, carvacrol, and E-caryophyllene, were identified, regardless of light conditions. The amount and composition of volatile compounds varied according to light intensity and quality. Low intensity (26 µmol m−2 s−1) increased γ-terpinene content (12.42%) and concomitantly decreased carvacrol (38.52%). Blue LED light showed higher production of carvacrol (48.11%).

Species-specific antimicrobial activity of essential oils and enhancement by encapsulation in mesoporous silica nanoparticles

Essential oils are volatile plant compounds that are biologically active and play an important role in natural plant protection. There are currently ∼3000 essential oils known, of which over 300 are commercially important for a variety of industries. In fact, the essential oil global market has been estimated to reach 13.94 billion USD with a demand of over 370,000 tons by 2024. These compounds have a wide variety of applications in agriculture, food and beverage, cosmetics, medicine, amongst other industries. A promising application of essential oils is as antimicrobials to target the vast number of diseases affecting crops. However, their volatile nature limits their effective use as free agents. To overcome this, we have investigated the use of mesoporous silica nanoparticles to protect essential oils from evaporation and degradation and to enhance their antimicrobial activity against bacterial phytopathogens. Silica nanoparticles were used due to their potential to be produced at an industrial scale and their biocompatibility. As a proof of concept, we evaluated 41 essential oils against bacterial phytopathogen Pseudomonas syringae pv. pisi, causative agent of pea bacterial blight. Additionally, we compared the effect of such essential oils against Pectobacterium carotovorum subsp. carotovorum and Pseudomonas fluorescens. Two of the most effective antimicrobials, cinnamon (Cinnamomum zeylanicum) and mustard (Brassica nigra) oils, were able to inhibit bacterial growth after 24 h at a concentration as low as 0.016% (v/v). Besides efficacy, the species-specificity of essential oils was demonstrated with >67% of oils tested displaying specificity towards pathogenic P. syringae pv. pisi over non-pathogenic Pseudomonas fluorescens. Furthermore, the encapsulation of essential oils into mesoporous silica nanoparticles (MSNPs) as a means of extending and improving their antimicrobial effect was found to enable a 10-fold increase in potency compared to the free essential oil. Cinnamaldehyde immobilised onto MSNPs proved to be the most effective antimicrobial, eliminating >99.8%, >99.9%, and >95% bacterial growth of P. fluorescens, P. syringae pv. pisi and P. carotovorum subsp. carotovorum, respectively. This system has the potential to be used to treat and prevent bacterial infections in crops and to enable a more controlled and effective exploitation of volatile compounds as antimicrobials.

Volatile compounds profile changes from unripe to ripe fruits of Brazilian pepper (Schinus terebinthifolia Raddi)

This study aimed to compare the chemistry of plant volatile compounds (VCs) of the unripe and ripe fruit of Schinus terebinthifolia Raddi by gas chromatography/mass spectrometry (GC–MS). The yield, density, qualitative and quantitative composition, chemical diversity and equitability of the samples were compared. Both essential oils had residual traces of sesquiterpenoids and are mainly composed of monoterpenes which represent 80.65% of the total compounds identified for unripe fruits oil and 81.36% for ripe oil. The predominant substances of the VCs of the ripe fruits were δ-3-carene (46.67%) and α-pinene (14.98%), whereas the major compounds of that of the unripe fruit were limonene (37.49%) and δ-3-carene (19.98%). Only 13 of the 98 substances studied were common to both samples. The yield of essential oil from the fruit of S. terebinthifolia as the raw material for extraction, ripe fruit were twice more productive than the unripe fruit ones. In the ripe fruit, δ-3 carene and α-pinene are possible chemical markers of this species, while limonene can be used as a substance characteristic for unripe fruit. The results suggest differences in chemical composition between the ripe and unripe fruit of S. terebinthifolia according to its ripening stage and raise subsidies for a plant of natural extractivism of Brazilian Pepper.

Shared weapons in fungus-fungus and fungus-plant interactions? Volatile organic compounds of plant or fungal origin exert direct antifungal activity in vitro

Abstract Fungi emit a diverse blend of volatile organic compounds (VOCs) that mediate multiple fungus-fungus interactions. Plants emit VOCs as well, which can serve as resistance-inducing signals but might also act as direct resistance agents. We screened 22 VOCs that are emitted from infected plants for putative inhibitory effects on three fungal phytopathogens: Colletotrichum lindemuthianum, Fusarium oxysporum and Botrytis cinerea. The growth of all three fungi was significantly inhibited when the mycelia were exposed to an atmosphere containing nonanal, (+)-carvone, citral, trans-2-decenal, L-linalool, or nerolidol. Eugenol completely inhibited the growth of all three fungi, and 1-octen-3-ol, nonanal, 2,6-dimethyl-2,4,6-octatriene, citral, α-terpineol and trans-2-decanal inhibited at least one fungus completely. Most of these VOCs are also emitted from fungi. We conclude that antagonistic fungus-fungus and plant-fungus interactions might share common mechanisms and that plant VOCs can function as resistance agents that fungal pathogens must overcome for successful infection.

Rice terpene synthase 18 (OsTPS18) encodes a sesquiterpene synthase that produces an antibacterial (E)-nerolidol against a bacterial pathogen of rice

Plant volatile compounds, including terpenes, are known to be involved in the rice defense system. In the present analysis of a terpene synthase, OsTPS18, in rice, we found that OsTPS18 was localized in the cytoplasm and synthesized the sesquiterpenes (E)-nerolidol and (E)-β-farnesene. The amounts of (E)-nerolidol and (E)-β-farnesene increased after jasmonic acid (JA) treatment. (E)-Nerolidol had significant antibacterial activity against Xanthomonas oryzae pv. oryzae (Xoo). These results suggest that (E)-nerolidol plays an important role in JA-induced resistance against Xoo and that it functions as an antibacterial compound in rice.

Can Plant Defence Mechanisms Provide New Approaches for the Sustainable Control of the Two-Spotted Spider Mite Tetranychus urticae?

Tetranychus urticae (T. urticae) Koch is a cosmopolitan, polyphagous mite which causes economic losses in both agricultural and ornamental plants. Some traits of T. urticae hamper its management, including a short life cycle, arrhenotokous parthenogenesis, its haplodiploid sex determination system, and its extraordinary ability to adapt to different hosts and environmental conditions. Currently, the use of chemical and biological control are the major control methods used against this mite. In recent years, some studies have focused on plant defence mechanisms against herbivores. Various families of plant compounds (such as flavonoids, glucosinolates, or acyl sugars) have been shown to behave as acaricides. Plants can be induced upon appropriate stimuli to increase their resistance against spider mites. This knowledge, together with the understanding of mechanisms by which T. urticae detoxifies and adapts to pesticides, may complement the control of this pest. Herein, we describe plant volatile compounds (VOCs) with repellent activity, and new findings about defence priming against spider mites, which interfere with the T. urticae performance. The use of VOCs and defence priming can be integrated into current management practices and reduce the damage caused by T. urticae in the field by implementing new, more sustainable crop management tools.

Design of an Attractant for Empoasca onukii (Hemiptera: Cicadellidae) Based on the Volatile Components of Fresh Tea Leaves.

The tea leafhopper, Empoasca onukii Matsuda, is a serious pest of the tea plant. E. onukii prefers to inhabit vigorously growing tender tea leaves. The host selection of E. onukii adults may be associated with plant volatile compounds (VOCs). We sought to identify potentially attractive VOCs from tea leaves at three different ages and test the behavioral responses of E. onukii adults to synthetic VOC blends in the laboratory and field to aid in developing an E. onukii adult attractant. In darkness, the fresh or mature tea leaves of less than 1-mo old could attract more leafhoppers than the mature branches (MB) that had many older leaves (leaf age >1 mo). Volatile analysis showed that the VOC composition of the fresh leaves was the same as that of the mature leaves, but linalool and indole were not at detectable levels in VOCs from the MB. Moreover, the mass ratio differed for each common volatile in the three types of tea leaves. When under competition with volatiles from the MB, the leafhoppers showed no significant tropism to each single volatile but could be attracted by the synthetic volatile blend imitating the fresh leaves. With the removal of some volatile components, the effective synthetic volatile blend was mixed with (Z)-3-hexen-1-ol, (Z)-3-hexenyl acetate, and linalool at a mass ratio of 0.6:23:12.6. These three volatiles may be the key components for the host selection of E. onukii adults and could be used as an attractant in tea gardens.

Effects of intraspecific and intra-individual differences in plant quality on preference and performance of monophagous aphid species.

Plant chemistry is one of the main drivers of herbivore distribution. Monophagous herbivore species are highly specialized, but even within their only host species the chemistry varies. The herbivore's choice is initially mainly guided by volatile plant compounds. Once on the plant, particularly for aphids the phloem quality affects their performance. However, little is known about the intraspecific and intra-individual variation in phloem sap and their influences on monophagous aphids. To determine potential mechanisms involved in aphid colonization, we tested the effects of intraspecific chemical variation in Tanacetum vulgare, which produces different chemotypes, on the preference of two monophagous aphid species. Moreover, we measured the performance of the aphids on different plant parts (stem close to the inflorescence, young and old leaves) of these chemotypes and analyzed their phloem sap composition. Both species preferred the β-thujone (THU) over the trans-carvyl acetate (CAR) chemotype in dual-choice assays. Survival of Macrosiphoniella tanacetaria was neither affected by intraspecific nor intra-individual variation, whereas the reproduction was highest on stems. In contrast, Uroleucon tanaceti survived and reproduced best on old leaves of the preferred chemotype. The sugar, organic acid and amino acid composition pronouncedly differed between phloem exudates of different plant parts, but less between chemotypes. Unexpectedly, high concentrations of amino acids did not necessarily enhance aphid performance. These different performance optima may cause niche differentiation and, therefore, enable co-existence. In conclusion, the tremendous variation in plant chemistry even within one species can affect the distribution of highly specialized aphids at various scales aphid species-specifically.

Does Background Odor in Tea Gardens Mask Attractants? Screening and Application of Attractants for Empoasca onukii Matsuda.

Plant volatiles help herbivores to locate their hosts, and therefore, they could be used to help develop pesticide-free pest management strategies. To develop an attractant for tea leafhopper (Empoasca onukii), we screened nine tea plant volatile compounds for their attractiveness using Y-tube olfactometer assays. Results indicated that tea leafhoppers significantly preferred ocimene, limonene, (Z)-3-hexenol, and (Z)-3-hexenyl acetate over clean air. These compounds were combined in a blend which lost its attractiveness at concentrations below 10-2 g/ml in liquid paraffin. In field tests, the blend was attractive to leafhoppers only in autumn, but not in summer. Analyses of the tea field background odor showed that all four components of the blend were present at much higher concentrations in summer (0.05-0.001 ng/liter) than in autumn (~10- to 25-fold lower). In field Y-tube bioassays, compared with the tea field background odor, the blend was attractive at a concentration of 10-1 g/ml in liquid paraffin, but not at 10-2 g/ml. These results suggest that field background odor can disrupt the attractiveness of an attractant based on plant volatiles to herbivores.

Antennal and behavioral responses of Mythimna separata (Walker) to three plant volatiles.

The oriental armyworm, Mythimna separata, is distributed widely in eastern Asia and Australia. The response of M.separata to 27 compounds identified from plant volatiles was determined from electroantennography (EAG) and wind tunnel results, which allowed an evaluation of the possible plant volatile compounds. The highest EAG values of males were elicited by trans-2,cis-6-nonadienal, and virgin females by benzyl alcohol. The amplitude in EAG dose-response was in the range of 0.24 to 2.87mV. In the wind tunnel bioassays, significantly more females showed behavioral responses to wilting leaves and headspace collection of Pterocarya stenoptera rather than control. In addition, significantly more females flew upwind with beta-ocimene compared with the control. The number of females that landed at the source with cis-3-hexen-1-ol, phenylethyl alcohol, trans-2-nonenal, and 2-pentylfuran was significantly different from the number that moved towards control.

Functional analysis of female-biased odorant binding protein 6 for volatile and nonvolatile host compounds in Adelphocoris lineolatus (Goeze).

The polyphagous mirid bug Adelphocoris lineolatus relies heavily on olfactory cues to track suitable host plants. Thus, a better understanding of the molecular basis of its olfactory detection could contribute to the development of effective pest management strategies. In the present study, we report the expression profile of the odorant binding protein gene A.lineolatus odorant binding protein 6 (AlinOBP6). Quantitative real-time PCR experiments suggest that AlinOBP6 is female adult antennae-biased. Cellular immunolocalization analyses show that AlinOBP6 is highly expressed in the lymph of both multiporous sensilla basiconica and uniporous sensilla chaetica. A ligand binding analysis showed that recombinant AlinOBP6 not only bound tightly to host plant volatile compounds but also to nonvolatile compounds. Homology modelling and molecular docking analyses confirmed these unusual ligand binding profiles and revealed that the amino acid residues involved in the recognition of volatile and nonvolatile compounds are distinct. The results of our study are the first to suggest that an antenna- and female-biased OBP in an hemipteran insect is expressed in both olfactory and gustatory sensilla as a mechanism to respond to volatile and nonvolatile host compounds. These findings warrant further research into the molecular mechanisms of chemosensation for mirid bugs in responsive to host plant location.

Tomato Flavor: Lost and Found?

Tomato Flavor: Lost and Found?

Ontogenetic shift in plant-related cognitive specialization by a mosquito-eating predator

Evarcha culicivora, an East African salticid spider, is a mosquito specialist and it is also a plant specialist, with juveniles visiting plants primarily for acquiring nectar meals and adults visiting plants primarily as mating sites. The hypothesis we consider here is that there are ontogenetic shifts in cognition-related responses by E. culicivora to plant odour. Our experiments pertain to cross-modality priming effects in three specific contexts: executing behaviour that we call the ‘visual inspection of plants’ (Experiment 1), adopting selective visual attention to specific visual targets (Experiment 2) and becoming prepared to respond rapidly to specific visual targets (Experiment 3). Our findings appear not to be a consequence of salient odours in general elevating E. culicivora’s motivation to respond to salient visual stimuli. Instead, effects were specific to particular odours paired with particular visual targets, with the salient volatile plant compounds being caryophyllene and humulene. We found evidence that prey odour primes juveniles and adults to respond to seeing specifically prey, mate odour primes adults to respond to seeing specifically mates and plant odour primes juveniles to respond to seeing specifically flowers. However, plant odour appears to prime adults to respond to seeing specifically a mate associated with a plant.

Evaluation of antibacterial potential and toxicity of plant volatile compounds using new broth microdilution volatilization method and modified MTT assay

With aim to develop effective proof-of-concept approach which can be used in a development of new preparations for the inhalation therapy, we designed a new screening method for simple and rapid simultaneous determination of antibacterial potential of plant volatiles in the liquid and the vapour phase at different concentrations. In addition, EVA (ethylene vinyl acetate) capmat™ as vapour barrier cover was used as reliable modification of thiazolyl blue tetrazolium bromide (MTT) assay for cytotoxicity testing of volatiles on microtiter plates. Antibacterial activity of carvacrol, cinnamaldehyde, eugenol, 8-hydroxyquinoline, thymol and thymoquinone was determined against Haemophilus influenzae, Staphylococcus aureus, and Streptococcus pneumoniae using new broth microdilution volatilization method. The cytotoxicity of these compounds was evaluated using MTT test in lung fibroblast cells MRC-5. The most effective antibacterial agents were 8-hydroxyquinoline and thymoquinone with the lowest minimum inhibitory concentrations (MICs) ranging from 2 to 128μg/mL, but they also possessed the highest toxicity in lung cell lines with half maximal inhibitory concentration (IC50) values 0.86–2.95μg/mL. The lowest cytotoxicity effect was identified for eugenol with IC50 295.71μg/mL, however this compound produced only weak antibacterial potency with MICs 512–1024μg/mL. The results demonstrate validity of our novel broth microdilution volatilization method, which allows cost and labour effective high-throughput antimicrobial screening of volatile agents without need of special apparatus. In our opinion, this assay can also potentially be used for development of various medicinal, agricultural, and food applications that are based on volatile antimicrobials.

A Background of a Volatile Plant Compound Alters Neural and Behavioral Responses to the Sex Pheromone Blend in a Moth.

Recognition of intra-specific olfactory signals within a complex environment of plant-related volatiles is crucial for reproduction in male moths. Sex pheromone information is detected by specific olfactory receptor neurons (Phe-ORNs), highly abundant on the male antenna. The information is then transmitted to the pheromone processing macroglomerular complex (MGC) within the primary olfactory center, the antennal lobe, where it is processed by local interneurons and projection neurons. Ultimately a behavioral response, orientation toward the pheromone source, is elicited. Volatile plant compounds (VPCs) are detected by other functional types of olfactory receptor neurons (ORNs) projecting in another area of the antennal lobe. However, Phe-ORNs also respond to some VPCs. Female-produced sex pheromones are emitted within a rich environment of VPCs, some of which have been shown to interfere with the detection and processing of sex pheromone information. As interference between the different odor sources might depend on the spatial and temporal features of the two types of stimuli, we investigated here behavioral and neuronal responses to a brief sex pheromone blend pulse in a VPC background as compared to a control background in the male noctuid moth Agrotis ipsilon. We observed male orientation behavior in a wind tunnel and recorded responses of Phe-ORNs and MGC neurons to a brief sex pheromone pulse within a background of individual VPCs. We also recorded the global input signal to the MGC using in vivo calcium imaging with the same stimulation protocol. We found that VPCs eliciting a response in Phe-ORNs and MGC neurons masked responses to the pheromone and decreased the contrast between background odor and the sex pheromone at both levels, whereas α-pinene did not interfere with first order processing. The calcium signal produced in response to a VPC background was tonic, lasting longer than the VPC stimulus duration, and masked entirely the pheromone response. One percent heptanal and linalool, in addition to the masking effect, caused a clear delay in responses of MGC neurons to the sex pheromone. Upwind flight toward the pheromone in a wind tunnel was also delayed but otherwise not altered by different doses of heptanal.

Candidate pheromone receptors of codling moth Cydia pomonella respond to pheromones and kairomones

Olfaction plays a dominant role in the mate-finding and host selection behaviours of the codling moth (Cydia pomonella), an important pest of apple, pear and walnut orchards worldwide. Antennal transcriptome analysis revealed a number of abundantly expressed genes related to the moth olfactory system, including those encoding the olfactory receptors (ORs) CpomOR1, CpomOR3 and CpomOR6a, which belong to the pheromone receptor (PR) lineage, and the co-receptor (CpomOrco). Using heterologous expression, in both Drosophila olfactory sensory neurones and in human embryonic kidney cells, together with electrophysiological recordings and calcium imaging, we characterize the basic physiological and pharmacological properties of these receptors and demonstrate that they form functional ionotropic receptor channels. Both the homomeric CpomOrco and heteromeric CpomOrco + OR complexes can be activated by the common Orco agonists VUAA1 and VUAA3, as well as inhibited by the common Orco antagonists amiloride derivatives. CpomOR3 responds to the plant volatile compound pear ester ethyl-(E,Z)-2,4-decadienoate, while CpomOR6a responds to the strong pheromone antagonist codlemone acetate (E,E)-8,10-dodecadien-1-yl acetate. These findings represent important breakthroughs in the deorphanization of codling moth pheromone receptors, as well as more broadly into insect ecology and evolution and, consequently, for the development of sustainable pest control strategies based on manipulating chemosensory communication.

Analytical method for metabolites involved in biosynthesis of plant volatile compounds

The progress in the successful techniques used for studying metabolites involved in the metabolic routes of plant volatiles is summarized.

Anti-biofilm potential of some plant volatile compounds against Proteus mirabilis

Anti-biofilm potential of some plant volatile compounds against Proteus mirabilis