Interactions of Opuntia ficus-indica with Dactylopius coccus and D. opuntiae (Hemiptera: Dactylopiidae) through the Study of Their Volatile Compounds.

Opuntia ficus-indica has always interacted with many phytophagous insects; two of them are Dactylopius coccus and D. opuntiae. Fine cochineal (D. coccus) is produced to extract carminic acid, and D. opuntiae, or wild cochineal, is an invasive pest of O. ficus-indica in more than 20 countries around the world. Despite the economic and environmental relevance of this cactus, D. opuntiae, and D. coccus, there are few studies that have explored volatile organic compounds (VOCs) derived from the plant-insect interaction. The aim of this work was to determine the VOCs produced by D. coccus and D. opuntiae and to identify different VOCs in cladodes infested by each Dactylopius species. The VOCs (essential oils) were obtained by hydrodistillation and identified by GC-MS. A total of 66 VOCs from both Dactylopius species were identified, and 125 from the Esmeralda and Rojo Pelón cultivars infested by D. coccus and D. opuntiae, respectively, were determined. Differential VOC production due to infestation by each Dactylopius species was also found. Some changes in methyl salicylate, terpenes such as linalool, or the alcohol p-vinylguaiacol were related to Dactylopius feeding on the cladodes of their respective cultivars. Changes in these VOCs and their probable role in plant defense mechanisms should receive more attention because this knowledge could improve D. coccus rearing or its inclusion in breeding programs for D. opuntiae control in regions where it is a key pest of O. ficus-indica.

Formulation study of PLGA in situ films for topical delivery of salicylates.

A film-forming system (FFS) represents a convenient topical dosage form for drug delivery. In this study, a non-commercial poly(lactic-co-glycolic acid) (PLGA) was chosen to formulate an FFS containing salicylic acid (SA) and methyl salicylate (MS). This unique combination is advantageous from a therapeutic point of view, as it enabled modified salicylate release. It is beneficial from a technological perspective too, because it improved thermal, rheological, and adhesive properties of the in situ film. DSC revealed complete dissolution of SA and good miscibility of MS with the polymer. MS also ensures optimal viscoelastic and adhesive properties of the film, leading to prolonged and sustained drug release. The hydrolysis of MS to active SA was very slow at skin pH 5.5, but it apparently occurred at physiological pH 7.4. The film structure is homogeneous without cracks, unlike some commercial preparations. The dissolution study of salicylates revealed different courses in their release and the influence of MS concentration in the film. The formulated PLGA-based FFS containing 5 % SA and 10 % MS is promising for sustained and prolonged local delivery of salicylates, used mainly for keratolytic and anti-inflammatory actions and pain relief.

Efficacy of Herbivore-Induced Plant Volatile Methyl Salicylate in Evaluating the Seasonal Abundance of Herbivorous Thrips (Thysanoptera: Thripidae) in Sweet Pepper.

Herbivorous thrips that damage fruits, vegetables, field crops, and ornamentals are challenging to control using insecticides and need an integrated approach (IPM) for their management. Herbivore-induced plant volatiles (HIPVs) are semiochemical plants produced to attract natural enemies (NEs) of arthropod herbivores. Sex pheromones are animal-based semiochemicals that can attract males or females of conspecifics. The HIPV methyl salicylate (MS) is used in IPM to attract NEs. We conducted field experiments in 2018-2019 in Tennessee to study the efficacy of MS and the aggregation pheromone neryl (S)-2-methylbutanoate (NMB), which attracts Frankliniella occidentalis (FO), a dominant pest of many crops, in attracting thrips using sweet peppers. We found a significantly higher number of thrips in traps baited with MS than in the traps containing NMB when compared with a no-lure control. All treatments caught only one thrips species, Frankliniella tritici (FT), a significant pest of young cotton. It can also lower the abundance of FO in other crops. Our findings show that although FO was not found in the study location in Tennessee, traps baited with MS are suitable for managing FT and reducing FO in susceptible crops by increasing FT and attracting NEs to crop productions that use IPM-based management practices.

Molecular evolution of methylesterase family genes and the BnMES34 is a positive regulator of Plasmodiophora brassicae stress response in Arabidopsis.

Methylesterases (MES) are involved in hydrolysis of carboxylic esters, which have substantial roles in plant metabolic activities and defense mechanisms. This study aimed to comprehensively investigate Brassica napus BnMESs and characterize their role in response to Plasmodiophora brassicae stress. Forty-four BnMES members were identified and categorized into three groups based on their phylogenetic relationships and structural similarities. Through functional predictions in the promoter regions and analysis of RNA-Seq data, BnMES emerged as pivotal in growth, development, and stress responses to B. napus, particularly BnMES34, was strongly induced in response to P. brassicae infection. Gene Ontology analyses highlighted BnMES34's role in regulation of plant disease resistance responses. Furthermore, overexpression of BnMES34 in A. thaliana exhibited milder clubroot symptoms, and reduced disease indices, suggesting positive regulatory role of BnMES34 in plant's response to P. brassicae stress. Molecular docking and enzyme activity verification indicated that BnMES34 has the ability to generate salicylic acid via methyl salicylate, and further experimentally validated in vivo. This discovery indicates that the overexpression of BnMES34 in Arabidopsis confers resistance against clubroot disease. Overall, our research suggests that BnMES34 has a beneficial regulatory role in enhancing stress resistance to P. brassicae in B. napus.

Methyl-salicylate (MeSA)-mediated airborne defence.

Stressed plants emit a variety of chemicals into the environment, leading to increased pest resistance in neighbouring plants but the genetic and molecular mechanisms of the emissions remain obscure. Recently, Gong et al. identified novel methyl salicylate (MeSA)-mediated airborne defence that confers resistance to neighbouring plants against aphids and viruses.

A tug-of-war to control plant emission of an airborne alarm signal.

Aphids represent a major threat to crops. Hundreds of different viruses are aphid-borne. Upon aphid attack, plants release volatile organic compounds (VOCs) as airborne alarm signals to turn on the airborne defense (AD) of neighboring plants, thereby repelling aphids as well as reducing aphid fitness and virus transmission. This phenomenon provides a critical community-wide plant protection to fend off aphids, but the underlying molecular basis remains undetermined for a long time. In a recent article, Gong et al. established the NAC2-SAMT1 module as the core component regulating the emission of methyl-salicylate (MeSA), a major component of VOCs in aphid-attacked plants. Furthermore, they showed that SABP2 protein is critical for the perception of volatile MeSA signal by converting MeSA to Salicylic Acid (SA), which is the cue to elicit AD against aphids at the community level. Moreover, they showed that multiple viruses use a conserved glycine residue in the ATP-dependent helicase domain in viral proteins to shuttle NAC2 from the nucleus to the cytoplasm for degradation, leading to the attenuation of MeSA emission and AD. These findings illuminate the functional roles of key regulators in the complex MeSA-mediated airborne defense process and a counter-defense mechanism used by viruses, which has profound significance in advancing the knowledge of plant-pathogen interactions as well as providing potential targets for gene editing-based crop breeding.

Human Skin Drug Metabolism: Relationships between Methyl Salicylate Metabolism and Esterase Activities in IVPT Skin Membranes.

The presence of esterase enzymes in human skin and their role in drug metabolism has been reported, but their distribution in the various skin layers and the relative contributions of those layers to metabolism is poorly defined. To gain further insight into esterase distribution, we performed in vitro skin permeation of a commercial 28.3% methyl salicylate (MeSA) cream (Metsal™) in Franz diffusion cells, using a range of human skin membranes, all from the same donor. The membranes were viable epidermis separated by a dispase II enzymatic method, heat separated epidermis, dermatomed skin, and dermis separated by a dispase II enzymatic method. Methyl salicylate and its metabolite, salicylic acid (SA), were measured by high-performance liquid chromatography. Alpha naphthyl acetate and Hematoxylin and Eosin staining provided qualitative estimations of esterase distribution in these membranes. The permeation of methyl salicylate after 24 h was similar across all membranes. Salicylic acid formation and permeation were found to be similar in dermatomed skin and dermis, suggesting dermal esterase activity. These results were supported by the staining studies, which showed strong esterase activity in the dermal-epidermal junction region of the dermis. In contrast with high staining of esterase activity in the stratum corneum and viable epidermis, minimal stained and functional esterase activity was found in heat-separated and dispase II-prepared epidermal membranes. The results are consistent with dispase II digesting hemidesmosomes, penetrating the epidermis, and affecting epidermal esterases but not those in the dermis. Accordingly, whilst the resulting dispase II-generated dermal membranes may be used for in vitro permeation tests (IVPT) involving esterase-based metabolic studies, the dispase II-generated epidermal membranes are not suitable for this purpose.

Small RNAs contribute to citrus Huanglongbing tolerance by manipulating methyl salicylate signaling and exogenous methyl salicylate primes citrus groves from emerging infection.

Citrus production is severely threatened by the devastating Huanglongbing (HLB) disease globally. By studying and analyzing the defensive behaviors of an HLB-tolerant citrus cultivar 'Shatangju', we discovered that citrus can sense Candidatus Liberibacter asiaticus (CLas) infection and induce immune responses against HLB, which can be further strengthened by both endogenously produced and exogenously applied methyl salicylate (MeSA). This immune circuit is turned on by an miR2977-SAMT (encoding a citrus Salicylate [SA] O-methyltransferase) cascade, by which CLas infection leads to more in planta MeSA production and aerial emission. We provided both transgenic and multi-year trail evidences that MeSA is an effective community immune signal. Ambient MeSA accumulation and foliage application can effectively induce defense gene expression and significantly boost citrus performance. We also found that miRNAs are battle fields between citrus and CLas, and about 30% of the differential gene expression upon CLas infection are regulated by miRNAs. Furthermore, CLas hijacks host key processes by manipulating key citrus miRNAs, and citrus employs miRNAs that coordinately regulate defense-related genes. Based on our results, we proposed that miRNAs and associated components are key targets for engineering or breeding resistant citrus varieties. We anticipate that MeSA-based management, either induced expression or external application, would be a promising tool for HLB control.

A mosaic of endogenous and plant-derived courtship signals in moths.

Insects rely on olfaction to guide a wide range of adaptive behaviors, including mate and food localization, mate choice, oviposition site selection, kin recognition, and predator avoidance.1 In nocturnal insects, such as moths2 and cockroaches,3 mate finding is stimulated predominantly by long-range species-specific sex pheromones, typically emitted by females. During courtship, at close range, males in most moth species emit a blend of pheromone compounds from an everted, often large, pheromone gland. While long-distance communication with sex pheromones has been remarkably well characterized in thousands of moth species,2,4 close-range chemosensory sexual communication remains poorly understood. We reveal that in the moth Chloridea virescens, the male pheromone consists of three distinct classes of compounds: de novo biosynthesized alcohols, aldehydes, acetates, and carboxylic acids that resemble the female's emissions; newly identified compounds that are unique to the male pheromone, such as aliphatic polyunsaturated hydrocarbons; and sequestered plant secondary compounds and hormone derivatives, including methyl salicylate (MeSA). Thus, males employ a mosaic pheromone blend of disparate origins that may serve multiple functions during courtship. We show that two olfactory receptors in female antennae are tuned to MeSA, which facilitates female acceptance of the male. Because MeSA is emitted by plants attacked by pathogens and herbivores,5 the chemosensory system of female moths was likely already tuned to this plant volatile, and males appear to exploit the female's preadapted sensory bias. Interestingly, while female moths (largely nocturnal) and butterflies (diurnal) diverged in their use of sensory modalities in sexual communication,6 MeSA is used by males of both lineages.

Characterization of the key odor-active compounds in different aroma types of Fu brick tea using HS-SPME/GC-MSO combined with sensory-directed flavor analysis.

Fu brick tea (FBT) is popular for its unique 'fungal flower' aroma, however, its key odor-active compounds are essentially unknown. In this study, the odor-active compounds of "stale-fungal" aroma (CJX), "fresh-fungal" aroma (QJX), and "fermentation-fungal" aroma (FJX) types FBT were extracted and examined by headspace solid phase microextraction (HS-SPME) combined with gas chromatography-mass spectrometry (GC-MS) and gas chromatographyolfactometry (GC-O). A total of 43 volatile and 38 odor-active compounds were identified by these methods. Among them, the content of dihydroactindiolide (4596-13189 µg/L), (E)-linalool oxide (2863-6627 µg/L), and benzyl alcohol (4992-6859 µg/L) were highest. Aroma recombination experiments further verified that these odor-active compounds could be simulated the overall aroma profile of FBT successfully. Furthermore, omission experiments confirmed that 15, 20, and 15 key odor-active compounds in CJX, QJX, and FJX FBT, respectively. This study will provide a theoretical basis for comprehensively understanding the formation of characteristic aromas in FBT.

Chimaphila umbellata; a biotechnological perspective on the coming-of-age prince's pine.

Chimaphila umbellata has been studied for almost two centuries now, with the first paper exploring the phytochemistry of the plant published in 1860. Almost all contemporary studies focus on the biotechnological advances of C. umbellata including its utilization as a natural alternative in the cosmetic, food, biofuel, and healthcare industry, with a special focus on its therapeutic uses. This literature review critically investigates the significance and applications of secondary metabolites extracted from the plant and presses on the biotechnological approaches to improve its utilization. C. umbellata is home to many industrially and medicinally important phytochemicals, the majority of which belong to phenolics, sterols, and triterpenoids. Other important compounds include 5-hydroxymethylfurfural, isohomoarbutin, and methyl salicylate (the only essential oil of the plant). Chimaphilin is the characteristic phytochemical of the plant. This review focuses on the phytochemistry of C. umbellata and digs into their chemical structures and attributes. It further discusses the challenges of working with C. umbellata including its alarming conservation status, problems with in-vitro cultivation, and research and development issues. This review concludes with recommendations based on biotechnology, bioinformatics, and their crucial interface.

Exogenous Application of Methyl Salicylate Induces Defence in Brassica against Peach Potato Aphid Myzus persicae.

Plants use a variety of secondary metabolites to defend themselves against herbivore insects. Methyl salicylate (MeSA) is a natural plant-derived compound that has been used as a plant defence elicitor and a herbivore repellent on several crop plants. The aim of this study was to investigate the effect of MeSA treatment of Brassica rapa subsp. chinensis ('Hanakan' pak choi) on its interactions with peach potato aphids, Myzus persicae, and their natural enemy, Diaeretiella rapae. For this, we selected two concentrations of MeSA (75 mg/L and 100 mg/L). Our results showed that aphid performance was significantly reduced on plants treated with MeSA (100 mg/L). In a cage bioassay, the MeSA (100 mg/L)-treated plants showed lower adult survival and larviposition. Similarly, the MeSA (100 mg/L)-treated plants had a significantly lower aphid settlement in a settlement bioassay. In contrast, the M. persicae aphids did not show any significant difference between the MeSA (75 mg/L)-treated and control plants. In a parasitoid foraging bioassay, the parasitoid D. rapae also did not show any significant difference in the time spent on MeSA-treated and control plants. A volatile analysis showed that the MeSA treatment induced a significant change in volatile emissions, as high numbers of volatile compounds were detected from the MeSA-treated plants. Our results showed that MeSA has potential to induce defence in Brassica against M. persicae and can be utilised in developing sustainable approaches for the management of peach potato aphids.

Biotransformation and metabolism of three methyl salicylate glycosides by gut microbiota in vitro.

MSTG-A, MSTG-B and Gualtherin are three natural methyl salicylate glycosides isolated from Dianbaizhu (Gaultheria leucocarpa var. yunnanensis), which is a traditional Chinese folk medicine widely used for the treatment of rheumatoid arthritis. They share the same mother nucleus with aspirin, exhibit similar activity and have fewer side effects. In this study, the incubation of MSTG-A, MSTG-B and gaultherin monomers with human fecal microbiota (HFM), microbiota in 4 intestinal segments (jejunum, ileum, cecal, and colon) and feces of rats in vitro was carried out to comprehensively and meticulously understand their metabolism by gut microbiota (GM) in the body. MSTG-A, MSTG-B and Gualtherin were hydrolyzed by GM to lose glycosyl moieties. The quantity and position of xylosyl moiety significantly affected the rate and extent of the three components being metabolized. The -glc-xyl fragments of these three components could not be hydrolyzed and broken by GM. In addition, the existence of terminal xylosyl moiety prolonged the degradation time. Different results appeared in metabolism of the three monomers by microbiota of different intestinal segments and feces due to the alternation of the species and abundance of microorganisms along the longitudinal axis of the intestinal lumen. Cecal microbiota had strongest degradation ability on these three components. The metabolic details of GM on MSTG-A, MSTG-B and Gualtherin were clarified in this study, providing data support and basis for clinical development and bioavailability improvement.

Factors Guiding the Orientation of Nymphal Spotted Lanternfly, Lycorma delicatula.

A mark-release-recapture experiment was conducted to evaluate the orientation of spotted lanternfly (SLF) Lycorma delicatula White (Hemiptera: Fulgoridae) nymphs when released equidistant between two trees. The experiment was repeated weekly for eight weeks in a heavily infested area with mature tree-of-heaven Ailanthus altissima (Mill.) Swingle (Sapindales: Simaroubaceae) planted in rows as ornamental street trees in Beijing, China. One tree in each pair received a methyl salicylate lure, and the lure was rotated between trees every week as it aged. Two additional independent variables for each tree were also analyzed: size and SLF population density. Marked-released SLF significantly chose trees with higher SLF population density over trees with lower density populations, and they also chose larger trees significantly more than smaller trees. Population density and tree size were better predictors of attraction than lures, but when those factors were controlled, SLF significantly chose trees with methyl salicylate lures over control trees for the first 4 weeks of lure life. Wild SLF distribution was assessed weekly, revealing strong aggregation in first and second instars that diminished with development to the third and fourth instars. Thus, nymphal SLF aggregate, and orientation is strongly guided by the presence of other SLF and tree size.

Aphid-repellent, ladybug-attraction activities, and binding mechanism of methyl salicylate derivatives containing geraniol moiety.

BACKGROUND: Aphids have been mainly controlled by traditional chemical insecticides, resulting in unamiable risk to the environment over the last decades. Push-pull strategy is regarded as a promising eco-friendly approach for aphid management through repelling aphid away and attracting their natural enemy. Methyl salicylate (MeSA), one of typical HIPVs (herbivore-induced plant volatiles), can repel aphids and attract ladybugs. Our previous studies discovered a new lead compound 3e, a salicylate-substituted carboxyl (E)-ß-farnesene derivative that had effective aphid-repellent activity. However, whether 3e has attractive activity to ladybug like MeSA is unknown. Meanwhile, to discover a new derivative for both deterring aphid and recruiting ladybug is meaningful for green control of aphids. RESULTS: Through the structural optimization of 3e, 14 new derivatives were designed and synthesized. Among them, compounds 4e and 4i had good aphid (Acyrthosiphon pisum) repellent activity, and compounds 3e, 4e and 4i had significant ladybug (Harmonia axyridis) attractive activity to males. Particularly, 4i exhibited manifest attractive effect on the females as well. Binding mechanism showed that 4i not only bound effectively with the aphid (Acyrthosiphon pisum) target ApisOBP9 thanks to its multiple hydrophobic interactions and hydrogen-bond, but also had strong binding affinity with ladybug target HaxyOBP15 due to the suitable steric space. Additionally, 4i displayed low toxicity to bee Apis mellifera. CONCLUSION: Compound 3e does exhibit attractive activity to male ladybug as MeSA. However, the new derivative 4i, with both pleasant aphid-repellent and ladybug-attraction activities, can be considered as a novel potential push-pull candidate for aphid control in sustainable agriculture. © 2022 Society of Chemical Industry.

New trends in methyl salicylate sensing and their implications in agriculture.

Methyl salicylate (MeSal) is an organic compound present in plants during stress events and is therefore a key marker for early plant disease detection. It has usually been detected by conventional methods that require bulky and costly equipment, such as gas chromatography or mass spectrometry. Currently, however, chemical sensors provide an alternative for MeSal monitoring, showing good performance for its determination in the vapour or liquid phase. The most promising concepts used in MeSal determination include sensors based on electrochemical and conductometric principles, although other technologies based on mass-sensitive, microwave, or spectrophotometric principles also show promise. The receptor elements or sensitive materials are shown to be part of the key elements in these sensing technologies. A literature survey identified a significant contribution of bioreceptors, including enzymes, odourant-binding proteins or peptides, as well as receptors based on polymers or inorganic materials in MeSal determination. This work reviews these concepts and materials and discusses their future prospects and limitations for application in plant health monitoring.

Expression and functional characterization of odorant-binding protein 2 in the predatory mite Neoseiulus barkeri.

Olfaction plays a crucial role for arthropods in foraging, mating, and oviposition. The odorant-binding protein (OBP) gene is considered one of the most important olfactory genes. However, little is known about its functions in predatory mites. Here, we used Neoseiulus barkeri, an important commercialized natural pest control, to explore the chemosensory characteristics of OBP. In this study, N. barkeri was attracted by methyl salicylate (MeSA) and showed higher crawling speeds under MeSA treatment. Then, we identified and cloned an OBP gene named Nbarobp2 and analyzed its expression profiles in the predatory mite. Nbarobp2 was 663 bp, was highly expressed in larval and nymphal stages, and was significantly upregulated in N. barkeri under MeSA treatment. Nbarobp2 encoded 202 amino acid residues with a molecular weight of 23 kDa (after removing the signal peptide). Sequence comparisons revealed that the OBPs in Arachnida shared 6 conserved cysteine sites, but were distinguishable from the OBPs of Insecta on the phylogenetic tree. RNA interference, Western blotting, and binding affinity assays further proved that Nbarobp2 was involved in volatile perception in predatory mites. This study shed light on the functional characteristics of OBPs in predatory mites, providing a new insight for better biological control.

An integrated in vitro approach on the enzymatic and antioxidant mechanisms of four commercially available essential oils (Copaifera officinalis, Gaultheria fragrantissima, Helichrysum italicum, and Syzygium aromaticum) traditionally used topically for their anti-inflammatory effects.

Introduction: Despite the increasing number of essential oils being reported on their potential therapeutic effects, some remain relatively unknown on their biological properties. That is the case of the essential oils obtained from copaiba (Copaifera officinalis L.), wintergreen (Gaultheria fragrantissima Wall.), everlasting (Helichrysum italicum (Roth) G.Don) and clove (Syzygium aromaticum (L.) Merr. & L.M.Perry), commonly labelled as being useful on the amelioration of conditions with an inflammatory background. Methods: To further broaden the current knowledge on the four essential oils, commercially available samples were approached on their effects upon a series of mediators that are involved on the inflammatory and oxidative response, both through in vitro cell-free and cell-based assays (5-lipoxygenase activity, lipid peroxidation, free radical and nitric oxide radical scavenging properties or tyrosinase inhibition). Results: The four oils proved to be active at some of the concentrations tested in most of the performed assays. Significant differences were found between the essential oils, S. aromaticum proving to tbe the most active, followed by G. fragrantissima against 5-lipoxygenase (5-LOX) and linoleic acid peroxidation, proving their potential use as antioxidants and anti-inflammatory agents. In fact, the IC50 value of S. aromaticum in the 5-LOX assay was 62.30 µg mL-1. Besides S. aromaticum efficiently scavenged superoxide radicals generated by xanthine/xanthine oxidase, displaying an IC50 value of 135.26 µg mL-1. The essential oil obtained from H. italicum exhibited a significant decrease in the nitric oxide levels on BV-2 cells, showing its potential as a cytoprotective agent against toxic damage. Copaiba oil ranked first as the most potent tyrosinase inhibitor, exhibiting an IC50 98.22 µg mL-1. Conclusion: More studies are needed to describe the essential oils properties, but these results confirm the potential of these essential oils as anti-inflammatory and antioxidant agents.

Exogenous Application of a Plant Elicitor Induces Volatile Emission in Wheat and Enhances the Attraction of an Aphid Parasitoid Aphidius gifuensis.

It is well known that plant elicitors can induce plant defense against pests. The herbivore-induced plant volatile (HIPV) methyl salicylate (MeSA), as a signaling hormone involved in plant pathogen defense, is used to recruit natural enemies to protect wheat and other crops. However, the defense mechanism remains largely unknown. Here, the headspace volatiles of wheat plants were collected and analyzed by gas chromatography-mass spectrometry (GC-MS), gas chromatography with electroantennographic detection (GC-EAD) and principal component analysis (PCA). The results showed that exogenous application of MeSA induced qualitative and quantitative changes in the volatiles emitted from wheat plants, and these changes were mainly related to Carveol, Linalool, m-Diethyl-benzene, p-Cymene, Nonanal, D-limonene and 6-methyl-5-Hepten-2-one. Then, the electroantennogram (EAG) and Y-tube bioassay were performed to test the physiological and behavioral responses of Aphidius gifuensis Ashmesd to the active volatile compounds (p-Cymene, m-Diethyl-benzene, Carveol) that identified by using GC-EAD. The female A. gifuensis showed strong physiological responses to 1 µg/µL p-Cymene and 1 µg/µL m-Diethyl-benzene. Moreover, a mixture blend was more attractive to female A. gifuensis than a single compound. These findings suggested that MeSA could induce wheat plant indirect defense against wheat aphids through attracting parasitoid in the wheat agro-ecosystem.

Solid Phase Microextraction-Multicapillary Column-Ion Mobility Spectrometry (SPME-MCC-IMS) for Detection of Methyl Salicylate in Tomato Leaves.

Methyl salicylate (MeSA) is a plant-signaling molecule that plays an essential role in the regulation of plant responses to biotic and abiotic pathogens. In this work, solid phase microextraction (SPME) and a multicapillary column (MCC) are coupled to ion mobility spectrometry (IMS) to detect MeSA in tomato leaves. The SPME-MCC-IMS method provides two-dimensional (2D) separation by both MCC and IMS, based on the retention and drift times. The effect of the IMS polarity on the separation efficiency of MCCs was also investigated. In the positive polarity, ionization of MeSA resulted in [MeSA + H]+ formation while, in the negative, deprotonated ions, [MeSA - H]-, and the O2- adduct ion, [MeSA + O2]-, were formed. In the real sample analysis, the negative polarity operation resulted in the suppression of many matrix molecules and thus in the reduction of interferences. Four different SPME fibers were used for head space analysis, and four MCC columns were investigated. In the negative polarity, complete separation was achieved for all of the MCCs columns. The limits of detection (LODs) of 0.1 µg mL-1 and linear range of 0.25-12 µg mL-1 were obtained for the measurement of MeSA in a standard solution (H2O/CH3OH, 50:50) by the SPME-IMS method with a 5 min extraction time using an SPME with a PDMS fiber, in the negative mode of IMS. The MeSA contents of fresh tomato leaves were determined as 1.5-9.8 µg g-1, 24-96 h after inoculation by tomato mosaic ringspot virus (ToRSV).