Behavioral Responses of Egg Parasitoids Telenomus remus and Trichogramma chilonis to Kairomonal cues released from Eggs and Wings of Fall Armyworm (Spodoptera frugiperda J.E. Smith)

Behavioral Responses of Egg Parasitoids Telenomus remus and Trichogramma chilonis to Kairomonal cues released from Eggs and Wings of Fall Armyworm (Spodoptera frugiperda J.E. 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Smith) Amit Kumar, S. Ramesh Babu, Devendra Jain, Heenashree Mansion This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8768024/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract The Fall Armyworm is a highly polyphagous, dreaded insect pest belonging to the order lepidoptera. During this study we, evaluated in behavioral bioassays, female Spodoptera frugiperda moths and two egg parasitoid species were tested for their responses to maize plant volatiles. Based on the combined findings, both Trichogramma chilonis and Telenomus remus egg parasitoids rely on combined plant and host egg cues for optimal attraction. Experienced females of both species show a stronger preference for odours from oviposited plants over plants or egg cues alone. Telenomus remus Trichogramma chilonis Egg cues Fall armyworm Behavioral bioassays Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 INTRODUCTION Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae), commonly known as the fall armyworm (FAW), is a notorious polyphagous insect indigenous to the Western Hemisphere's warmer climates (Kumar et al. 2025 ). The pest is reported to feed on a remarkably wide variety of hosts, with records indicating over 350 plant species within 76 families (Montezano et al. 2018 ). The urgent need for biological control of the fall armyworm arises from its devastating impact on crop yields and the severe limitations of sole reliance on chemical insecticides, which include rising resistance, environmental harm, and high costs for smallholder farmers (Ghafar et al. 2025 ). Host location is a central process in the life of parasitoid because finding suitable hosts is key for its reproduction success (Turlings and Erb 2018 ). At the beginning of foraging, parasitoids use different strategies and cues for locating their hosts, depending on the host stage. For parasitoids of eggs, the host location process may imply a higher challenge in comparison to larval or adult parasitoids, considering that eggs do not feed nor defecate, and thus do not emit long- range volatiles that can be used as cues by their natural enemies (Ngumbi et al. 2021). Several studies have investigated the role of herbivore-induced volatiles during the host searching by insect parasitoids using the tritrophic model of maize-Lepidopteran-parasitoids (Ortiz-Carreon et al. 2019 ). Plants emit volatile compounds that have a major impact on their environment (Tholl et al. 2018), and when undergoing herbivory, they produce specific Herbivore-Induced Plant Volatiles (HIPVs). These HIPVs function as indirect defense signals, providing olfactory cues that are critical for hymenoptera parasitoids to locate their hosts (Turlings et al. 1998). The induced changes in the plant's volatile blend convey information about the attacking herbivore and attract its natural enemies, thereby forming a key component of the plant's indirect defense response (Sharma et al. 2017 ).These volatiles act as signals to distal tissues of the plant itself, as well as to other organisms in the environment, providing information on the identity, location, and health of the plant (Karalija et al. 2023 ). The most common strategy used by egg parasitoids to find their hosts is exploiting chemical cues related to adult hosts, including sex, antisex, and aggregation pheromones, or host adult residues, such as genital secretions and scales that line the egg masses. A second strategy is to use of plant volatiles, commonly constitutive volatiles that are used as long- range chemical cues by egg parasitoids, though herbivory- induced plant volatiles may be more attractive cues for parasitoids by healthy or mechanically-damaged plants (Tepa-Yotto et al. 2021 ). The most common strategy used by egg parasitoids to find their hosts is exploiting chemical cues related to adult hosts, including sex, antisex, and aggregation pheromones, or host adult residues, such as genital secretions and scales that line the egg masses. METHODS AND MATERIALS FAW Culture The S. frugiperda larvae were collected from fall armyworm rearing laboratory, Department of Entomology, Rajasthan College of Agriculture, MPUAT, Udaipur. Larvae were maintained individually to avoid cannibalism. Later, larvae were taken to the insectary to be reared using an artificial diet under controlled conditions of temperature (26 ± 2°C), relative humidity (75 ± 5%), and a photoperiod of 12:12 h (light/dark) (Kumar et al. 2025 ). The pupae were kept in glass cages until adult emergence. Ten adults (5 females and 5 males) were place in Kraft paper bags for mating and oviposition. Adults were feed a 10% honey solution dispensed on cotton wool, and was kept under the conditions described above. S. frugiperda egg masses deposited in Kraft paper bags were collected daily. Parasitoids Culture The S. frugiperda larvae parasitized by parasitoid/s ( Telenomus remus and Trichogramma chilonis ) were collected from biological control laboratory, Department of Entomology, Rajasthan College of Agriculture, MPUAT, Udaipur and used as rearing stock for culturing wasps for bioassays. Three days-old adult parasitoids were placed in Petri dishes in a 1:1 sex ratio for mating. After mating, an egg mass of S. frugiperda offered to each Telenomus remus and Trichogramma chilonis female for oviposition. The egg masses were removed 24 h later, or until FAW larvae hatched; egg masses or neonate larvae were placed into containers, with artificial diet. Ten days later, S. frugiperda larvae were individually placed into transparent box until the emergence of adult of Telenomus remus and Trichogramma chilonis parasitoids. The parasitoids were fed with honey drops placed in the small container and kept under the laboratory conditions described above. Kairomones Collection from egg Surface To eliminate natural kairomones from the egg surface, 10 egg masses (0–24 hr old; about 2,500 eggs) taken from culture were washed twice in hexane (10 ml). The hexane egg wash rinsate was filtered using WhatmanTM No. 1 filter paper. The filtrates were kept at -20°C and utilized as stock for further diluting. FAW female moth scale collection and extraction Female fall armyworm scales were taken from freshly emerged (0–24 hrs old) laboratory-reared cultures by immobilising them at 0–2°C. The wings were removed from around 30 moths. The wing scales were extracted by shaking vigorously in a chilled shaker for 2 hours in 200 ml of analytical grade hexane and filtered through Whatman No.1 filter paper. The filtrate was kept at -20°C for Gas chromatography linked to mass spectrometry. Gas Chromatography Coupled with Mass Spectrophotometry (GC-MS) Using a GC-MS Agilent 7890B GC system with Mass Spectrometry (Agilent 5977 MSD), the chemical compositions of eggs and scales kairmone cues were examined. This was done in accordance with the method previously outlined by Jayanthi et al. ( 2021 ), with a few minor adjustments. The samples were examined using an Agilent J & W (HP-5 MS UI) capillary column that was 30 m long, 0. 250 mm in diameter, and had a film thickness of 0.25 µm. Helium was used as the carrier gas, and the thermal program was configured as previously mentioned. The flow rate was 1 mL/min. AMU varied between 40 and 450, while MS was operating in full scan mode (70 eV). 1.0 microlitre of the sample was inserted in splitless mode ratio (40 mL/min) with the injection temperature of 270ºC. The individual chemicals were initially identified by comparing GC retention time, Kovats index (C7 to C30 homologous series of n-alkenes as standard, Sigma-Aldrich; Kovats 1965) and comparing the mass spectra with spectral library NIST 14. The identified compounds were further authenticated and quantified by using a single point external standard quantification method by using authentic samples of standards (Ciotlaus et al. 2024) and the compounds for which synthetic standards are unavailable were identified tentatively based on NIST 14 spectral library. Bioassays To evaluate the behavioral responses of naive parasitoid to maize plants with and without eggs of fall armyworm, we were evaluated the responses of the parasitoids to maize plants with and without FAW eggs. Three seedlings were placed inside a collapsible mesh cage with 10 S. frugiperda gravid females 5–8 days old during one night to obtain plants with eggs. Moths were supplied with sugar water on cotton wool. The seedlings with 2–4 egg masses (oviposition-treated seedlings) were selected for the bioassays. For plants without eggs, seedlings of the same age as the oviposition treated seedlings but without FAW oviposition were used (egg-free seedlings). The following comparisons were performed: (1) Egg-free seedlings versus clean air, (2) Oviposition-treated seedlings versus clean air, (3) Egg-free seedlings plus 3–4 FAW egg masses oviposited on Kraft paper versus clean air, (4) Egg-free seedlings versus oviposition-treated seedlings (5) Egg-free seedlings plus 3–4 FAW egg masses oviposited on Kraft paper versus egg-free seedlings and (6) Egg-free seedlings plus 3–4 FAW egg masses oviposited on Kraft paper versus Kraft paper. The Kraft paper with the egg masses was placed on the seedlings leaves. Seedlings received eggs 12–14 h before the evaluation, wherase, behavioral responses of experienced parasitoid to maize plants with and without eggs of FAW, we were investigated that whether oviposition experience influences the responses of parasitoid females to maize plants with and without eggs. Parasitoid females with oviposition experience were obtained single females on seedlings previously oviposited by S. frugiperda so that they have contact with the plant and detect and parasitize eggs. The parasitoid females were removed 2 min later. The experienced females were tested the next day in the Y-tube olfactometer. The following comparisons were performed: (1) egg-free seedlings versus oviposition-treated seedlings, and (2) oviposition-treated seedlings versus 3–4 egg masses oviposited on Kraft paper. The bioassays were performed following the methodology described in the above section. While to test the behavioral responses of Naive parasitoid to egg masses of S. frugiperda , was evaluated that whether naive parasitoid females are attracted to host egg masses. Because Kraft paper was used as an oviposition substrate for FAW rearing; this material was included as a treatment in this experiment. The following comparisons were performed: (1) Kraft paper versus clean air, (2) 3–4 egg masses oviposited on Kraft paper versus clean air and (3) 3–4 egg masses oviposited on Kraft paper versus Kraft paper (Roque-Romero et al. 2020 ). RESULT Identification of Chemical Cues from Egg Surface Extracts The surface of eggs of FAW volatile emmisions were worked out using Gas Chromatography. The eggs surface emmited a total of 24 volatiles from 11 functional groups including sulfure compounds aldehydes, terpenes, esters, halogentad compounds, alcohols/ ethers, ketones, heterocytes, thioether, hydrocarborn, lignans and siloxanes, compounds have been identified. GCMS anaysis revealed siginificant differences among the functional chemical groups or classes of cues in surface of egg of S . frugiperda (Table 1 ). Among the chemical groups esters (Oxalic acid, cyclohexyl propyl ester, Oxalic acid, cyclohexyl isobutyl ester, Methyl allyl diglycolcarbonate, Carbonic acid, monoamide, N-pentyl-, decyl ester and Benzoic acid, 3,3'-[1,3-phenylenebis(carbonylimino)] bis-, dimethyl) were prodominent followed by hetereocycles (N-(2-Propynyl)aziridine, Thiophen-2-methylamine, N-(2-fluorophenyl)-, 2-(p-(Dimethylamino) phenyl) benzimidazole, 4H-1,2,4-Triazole, 4-ethyl- and 3-Pyridin-4-yl-2-(pyridin-4-yl-p-tolylaminomethyl)-3-p-tolylamino-propionitrile. Table 1 GC-MS profile of chemical cues of eggs of Spodoptera frugiperda Chemical Group Specific Compounds Area percentage 1. Esters Oxalic acid, cyclohexyl propyl ester 24.36 Oxalic acid, cyclohexyl isobutyl ester 2.14 Methyl allyl diglycolcarbonate 0.01 Carbonic acid, monoamide, N-pentyl-, decyl ester 13.50 Benzoic acid, 3,3'-[1,3-phenylenebis(carbonylimino)]bis-, dimethyl ester 0.009 2. Aldehydes Pentanal, 2,2-dimethyl- 19.66 3. Ketones/Epoxides Ethanone, 1-(3-methyloxiranyl)- 15.42 Bromoacetone 0.28 4. Ethers 4-Methyl-2-pentanol, methyl ether 1.08 3-Methoxy-3-methylbutanol 0.01 5. Thioethers Butanoic acid, 3-(methylthio)- 4.60 6. Halogenated Compounds 1,3-Butadiyne, 1,4-difluoro- 1.48 Hexanee, 2-bromo- 3.44 1(2H)-naphthalenone, 2-bromo-3,4-dihydro-6-methoxy- 2.54 7. Heterocycles/Nitrogenous N-(2-Propynyl)aziridine 0.59 Thiophen-2-methylamine, N-(2-fluorophenyl)- 1.69 2-(p-(Dimethylamino)phenyl)benzimidazole 0.02 4H-1,2,4-Triazole, 4-ethyl- 1.79 3-Pyridin-4-yl-2-(pyridin-4-yl-p-tolylaminomethyl)-3-p-tolylamino-propionitrile 0.12 8. Hydrocarbons Hepta-1,2,6-heptatriene 0.54 Butane, 2,2-dimethyl- 0.10 9. Cyclic Carbonates 1,3-Dioxol-2-one 1.57 10. Siloxanes Cyclotetrasiloxane, octamethyl- 0.02 11. Lignans Schizandrin 0.004 Identification of chemical compounds presents in headspace volatiles of wings cues of S. frugiperda A total of 22 volatiles from 10 functional groups, including esters, aldehydes, ketones,ethers,heterocycles, halogenated compounds, nitrogenous compounds,siloxanes, aromaticsand fluorinated compoundshave been identified from the surfaceof FAW wings(Table 4.39 and Fig. 4.64). The esters were the dominant group in the wings of S. frugiperda with five compounds, including Oxalic acid, cyclohexyl propyl ester, Oxalic acid, cyclohexyl isobutyl ester, Methyl allyl diglycolcarbonate, Carbonic acid, monoamide, N-pentyl-, decyl ester and Benzoic acid, 3,3'-[1,3-phenylenebis(carbonylimino)]bis-, dimethyl ester (Table 2 ). Table 2 GC-MS profile of chemical cues of wings of Spodoptera frugiperda Chemical Group Area percentage Compound Name 1. Esters 36.73 Oxalic acid, cyclohexyl propyl ester 4.27 Oxalic acid, cyclohexyl isobutyl ester 0.014 Oxalic acid, dineopentyl ester 1.06 Acetyl valeryl 0.04 Sulfurous acid, 2-ethylhexyl hexyl ester 1.14 Terephthalic acid, isobutyl 2,2,3,3,3-pentafluoropropyl ester 2. Aldehydes 30.67 Pentanal, 2,2-dimethyl- 3. Ketones/Epoxides 6.35 Ethanone, 1-(3-ethyloxiranyl)- 0.62 Acetyl bromide 4. Ethers 6.35 Ethane, 1,1-dimethoxy- 0.02 Butane, 1-(1-methylpropoxy)- 5. Heterocycles 0.12 Oxazole 0.56 (S)-(-)-1-Amino-2-(methoxymethyl)-pyrrolidine 0.11 N-[4-(1H-Benzoimidazol-2-yl)-phenyl]-acetamide 6. Halogenated Compounds 0.86 1-Buten-3-yne, 1-chloro-, (E)- 0.61 Cyclopropane, 2-bromo-1,1,3-trimethyl- 7. Nitrogenous Compounds 0.09 Cyclohexanee, nitro- 8. Siloxanes 8.57 Silicic acid, diethyl bis(trimethylsilyl) ester 9. Aromatics 0.52 3,4-Dimethoxy-N-[2-(2-methoxyphenoxy)ethyl]benzamide 10. Fluorinated Compounds 0.06 Bisphenol, O,O'-bis(pentafluoropropionyl)- Behavioural response of naïve female of Telenomus remus to maize plants with and without eggs of S. frugiperda in Y- olfactometer during single- choice bioassay The findings of present study revlealed that the responses of naïve female wasps of T. remus to odors from eggs free seedilings were significantly [χ 2 = 12.175; p < 0.0001], Ovipostion treated seedlings [χ 2 = 15.000; p < 0.0001], egg-free seedlings plus 3–4 FAW egg masses oviposited on Kraft paper [χ 2 = 10.756; p = 0.001] over clean air. The Oviposition treated seedling odour also significntly attrarctive to naïve female wasps [χ 2 = 20.000; p < 0.0001] over Egg- free seedlings. Egg-free seedlings plus 3–4 FAW egg masses oviposited on Kraft paper [χ 2 = 14.118; p < 0.0001] and Egg-free seedlings plus 3–4 FAW egg masses oviposited on Kraft paper [χ 2 = 12.593] significantly attracted over kraft paper. Present study indicated that the odors released from oviposited plants more attractive than normal plant and plant odour is attractive to parasitoids than clean air or other odors (Fig. 1 ). Behavioural response of experienced female of Telenomus remus to maize plants with and without eggs of S. frugiperda in Y- olfactometer during single- choice bioassay The experienced female T.remus were more attracted to odors from oviposited seedlings when compared to odors from egg-free seedlings [ χ 2 = 20.000; p < 0.0001] and Ovipostion treated seedlings [ χ 2 = 12.926; p < 0.0001] atrracted more females over 3–4 FAW egg masses oviposited on Kraft paper. Results of this experiment reveals that both plant volatiles and egg cues more attractive together (Fig. 2 ). Behavioural response of naïve female of Telenomus remus to egg masses of S. frugiperda in Y- olfactometer during single- choice bioassay The naïve females of T. remus had not show a significant preference for odors from Kraft paper and clean air [ χ 2 = 12.175; p = 0.005]. However, the T. remus females were attracted to odors emmited from egg masses oviposited in Kraft paper when compared to the clean air and kraft paper [ χ 2 = 15.000; p < 0.0001 and χ 2 = 10.756; p = 0.001, respectively]. The above mentioned findings are clearly evident that volatiles and cues released by plant and egg surface respectively are attaracted to egg parasitoids (Fig. 3 ). Behavioural response of naïve female of Trichogrmma chilonis to maize plants with and without eggs of S. frugiperda in Y- olfactometer during single- choice bioassay The findings of present study revlealed that the responses of naïve female wasps of T. chilonis to odors from eggs free seedilings were significantly [χ 2 = 15.556; p < 0.0001], Ovipostion treated seedlings [χ 2 = 6.667; p < 0.0001], egg-free seedlings plus 3–4 FAW egg masses oviposited on Kraft paper [χ 2 = 20.000; p = 0.001] over clean air. The Oviposition treated seedling odour also significntly attrarctive to naïve female wasps [χ 2 = 12.926; p < 0.0001] over Egg- free seedlings. Egg-free seedlings plus 3–4 FAW egg masses oviposited on Kraft paper [χ 2 = 13.003; p < 0.0001] and Egg-free seedlings plus 3–4 FAW egg masses oviposited on Kraft paper [χ 2 = 20.000; p < 0.0001] significantly attracted over kraft paper. Present study indicated that the odors released from oviposited plants more attractive than normal plant and plant odour is attractive to parasitoids than clean air or other odors (Fig. 4 ). Behavioural response of experienced female of Trichogrmma chilonis to maize plants with and without eggs of S. frugiperda in Y- olfactometer during single- choice bioassay The experienced female of T. chilonis were more attracted to odors from oviposited seedlings when compared to odors from egg-free seedlings [ χ 2 = 19.000; p < 0.0001] and Ovipostion treated seedlings [ χ 2 = 20.000; p < 0.0001] atrracted more females over 3–4 FAW egg masses oviposited on Kraft paper. Results of this experiment reveals that both plant volatiles and egg cues more attractive together (Fig. 5 ). Behavioural response of naïve female of Trichogrmma chilonis to egg masses of S. frugiperda in Y- olfactometer during single- choice bioassay The naïve females of T. chilonis had not show a significant preference for odors from Kraft paper and clean air [ χ 2 = 16.296; p < 0.0001]. However, the T. remus females were attracted to odors emmited from egg masses oviposited in Kraft paper when compared to the clean air and kraft paper [ χ 2 = 19.000; p < 0.0001 and χ 2 = 20.000; p = 0.001, respectively]. The above mentioned findings are clearly evident that volatiles and cues released by plant and egg surface respectively are attaracted to egg parasitoids (Fig. 6 ). Behavioural response of naive female of Telenomus remus to wing and egg cues extraction of S. frugiperda in Y- olfactometer during single- choice bioassay The behavioural response of naive female of Telenomus remus to wing and egg cues extraction of S. frugiperda in Y- olfactometer during single- choice bioassay. The egg and wing cues of S. frugiperda were signficantly more attractive when compared with hexan solvent [χ 2 = 3.951; p < 0.0001 and χ 2 = 20.000; p < 0.0001, respectively]. The females of T. remus were also significantly attracted to eggs cues than wing cues [χ 2 = 9.474; p < 0.0001] (Fig. 7 ). DISCUSSION During the present investigation we were found that the experienced female T. remus were more attracted to odours from oviposited seedlings and Ovipostion treated seedlings than egg masses oviposited on Kraft paper. The study also found that both plant volatiles and egg cues are more attractive together, indicating that both plant and egg surface odours are attracted to egg parasitoids. The present study also reveals that naïve female Trichogrmma chilonis wasps were significantly attracted to odours from egg-free seedlings, Ovipostion-treated seedlings, and egg-free seedlings plus 3–4 FAW egg masses oviposited on Kraft paper. The odours released from oviposited plants were more attractive than normal plants and plant odour was more attractive to parasitoids than clean air or other odours. Experienced female T. chilonis were more attracted to odours from oviposited seedlings and Oviposition-treated seedlings. The study also found that both plant volatiles and egg cues are more attractive together, with T. remus females being attracted to odours emitted from egg masses oviposited on Kraft paper. Present findings are closely associated with those of Roque-Romero et al. ( 2020 ) who reported that the C.insularis females responded to volatiles released by host egg masses, females (sex pheromones), and maize seedlings with or without FAW eggs. Females can distinguish between maize seedlings with and without FAW eggs due to the parasitoids' enhanced reactivity to host and maize seedling volatiles following their initial oviposition event. The authors presented theories to explain their findings, including the idea that FAW females limit herbivore-induced plant volatiles to avoid attracting parasitoids and predators. Some plant species change their volatile profile after herbivore oviposition, attracting parasitoids and predators from a distance (Nascimento et al. 2023). These chemical signals help the natural enemies of the herbivores locate infested plants, thus providing the plants with an indirect defense mechanism (Wang et al. 2025 ). This has been demonstrated in numerous studies showing that parasitoid wasps are drawn to the volatiles produced by plants under attack from their host insects (Nideesh et al. 2025). Similarly, the study found that C. insularis females are attracted to maize volatiles induced by larval feeding, but they are not the host stages of FAW larvae. The study confirmed that naïve C. insularis are attracted to FAW egg mass volatiles and female sex pheromone, as well as healthy maize seedling volatiles. The innate response to egg masses is not significant when the plant is present, or the parasitoid attraction increases with plant volatiles. Experienced female parasitoids can discriminate between previously oviposited maize seedlings and egg-free ones (Ortiz-Carreon et al. 2019 ). The volatiles obtained from larval and adult stages of Phthorimaea operculella , adult cues were observed to be more efficient in the alteration of foraging behavior of the targeted parasitoid, C. blackburni and the present study revealed that the C. blackburni preferred cues could be exploited to enhance the host searching efficiency during IPM releases. Several parasitoids are known to exploit these plant-provided cues to locate their hosts. One such parasitoid is the generalist Cotesia marginiventris , which is, among others, attracted to maize volatiles induced by caterpillar damage (Singh et al. 2019 ). This has been demonstrated in numerous studies showing that parasitoid wasps are drawn to the volatiles produced by plants under attack from their host insects (Nideesh et al. 2025). Similarly, Ooencyrtus spp. were found in plots with both A. contorta and S. montela larvae, indicating that the VOCs produced by feeding damage and larval saliva attract parasitoids (Davidson Lowe and Ali 2021; Gebreziher 2020 ; Moujahed et al. 2014 ). In conclusion, we evaluated that both plant volatiles and egg cues are more attractive together, indicating that both plant and egg surface odours are attracted to egg parasitoids. The present study also reveals that naïve female Trichogrmma chilonis wasps were significantly attracted to odours from egg-free seedlings, Ovipostion-treated seedlings, and egg-free seedlings plus 3–4 FAW egg masses oviposited on Kraft paper. The odours released from oviposited plants were more attractive than normal plants and plant odour was more attractive to parasitoids than clean air or other odours. Declarations Author Contribution The writing and technical corrections done by Devendra Jain and Heenashree Mansion. An experiment conducted by Amit Kumar and S. Ramesh babu Acknowledgments We are thankful Department of Entomology, Rajasthan College of Agriculture, Maharana Pratap University of Agriculture and Technology, Udaipur, Rajasthan (India) for technical assistance. The writing and technical correction done by ……….. 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Pro Nat Aca Scie 89:8399–8402 Wang J, Yi T, Wang M, Wei J, Yan W, Wen Y, Xu H (2025) Herbivore-induced maize volatiles: dual functions in repelling fall armyworm and attracting natural enemies. Pest Manage Scie 81:3674–3684 Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-8768024","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":589429708,"identity":"3ecc3141-1743-470b-b76d-4a01e7690e8e","order_by":0,"name":"Amit Kumar","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA8klEQVRIiWNgGAWjYFACxgYwxcbefPDBBzCDsJZGoB4DBj6eY8mGM0BamImzxoBBTiLHTJoHxCekhX/a4fYHH3f8kWOTyDE2tvm1TZ6PmYHxw8cc3Fokbic2Ns48Y2DMxvOs8HFu323DNmYGZsmZ2/BYA9TSzNtmkNjGnrzZOLfnNiNQCxszLx4t8iAtf0FaGBLMpC17btsT1GIA0sII0sKRYibN8ON2IkEthkAtM3vbjIF+AQZyb8Pt5DZmxma8fpG7nf7gw882OTn5dmBU/vhz23Y+kPHhIz7vowDGNjDZQKx6EPhDiuJRMApGwSgYKQAA9gBTgyKDVDsAAAAASUVORK5CYII=","orcid":"","institution":"Rajasthan College of Agriculture, Maharana Pratap University of Agriculture and Technology","correspondingAuthor":true,"prefix":"","firstName":"Amit","middleName":"","lastName":"Kumar","suffix":""},{"id":589429709,"identity":"8d68e600-a011-4af5-9c56-fdd32462f872","order_by":1,"name":"S. Ramesh Babu","email":"","orcid":"","institution":"Rajasthan College of Agriculture, Maharana Pratap University of Agriculture and Technology","correspondingAuthor":false,"prefix":"","firstName":"S.","middleName":"Ramesh","lastName":"Babu","suffix":""},{"id":589429710,"identity":"b39d13dc-2d96-4ad0-9567-1addfbb39569","order_by":2,"name":"Devendra Jain","email":"","orcid":"","institution":"Rajasthan College of Agriculture, Maharana Pratap University of Agriculture and Technology","correspondingAuthor":false,"prefix":"","firstName":"Devendra","middleName":"","lastName":"Jain","suffix":""},{"id":589429713,"identity":"14cd5422-ae8c-41b1-8965-a0987a6b564b","order_by":3,"name":"Heenashree Mansion","email":"","orcid":"","institution":"Agriculture University","correspondingAuthor":false,"prefix":"","firstName":"Heenashree","middleName":"","lastName":"Mansion","suffix":""}],"badges":[],"createdAt":"2026-02-02 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legend\u003c/p\u003e","description":"","filename":"Figures14.png","url":"https://assets-eu.researchsquare.com/files/rs-8768024/v1/64222abfdb1bf9347822fcee.png"},{"id":102746596,"identity":"66f4dc86-64a8-4c31-8d23-42784b9d59f4","added_by":"auto","created_at":"2026-02-16 08:58:31","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":24927,"visible":true,"origin":"","legend":"\u003cp\u003eSee image above for figure legend\u003c/p\u003e","description":"","filename":"Figures15.png","url":"https://assets-eu.researchsquare.com/files/rs-8768024/v1/dd5c43fd4f02579c90afcb3e.png"},{"id":102746978,"identity":"6981ec65-4e46-417b-8f43-87408b2bbcef","added_by":"auto","created_at":"2026-02-16 09:03:17","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":28723,"visible":true,"origin":"","legend":"\u003cp\u003eSee image above for figure legend\u003c/p\u003e","description":"","filename":"Figures16.png","url":"https://assets-eu.researchsquare.com/files/rs-8768024/v1/c03a674e2d5c7e0f606fc807.png"},{"id":102539917,"identity":"7437aacc-05a5-4893-a108-4edc99a0fb4f","added_by":"auto","created_at":"2026-02-12 18:44:49","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":27429,"visible":true,"origin":"","legend":"\u003cp\u003eSee image above for figure legend\u003c/p\u003e","description":"","filename":"Figures17.png","url":"https://assets-eu.researchsquare.com/files/rs-8768024/v1/45657f4e8c217c049052a764.png"},{"id":103342317,"identity":"d2efd7cb-30ae-4cb3-bd1c-fc91fdacd589","added_by":"auto","created_at":"2026-02-24 15:41:48","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1584144,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8768024/v1/14a47ee3-b602-4d40-8dec-659cc5f70dbf.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Behavioral Responses of Egg Parasitoids Telenomus remus and Trichogramma chilonis to Kairomonal cues released from Eggs and Wings of Fall Armyworm (Spodoptera frugiperda J.E. Smith)","fulltext":[{"header":"INTRODUCTION","content":"\u003cp\u003e \u003cem\u003eSpodoptera frugiperda\u003c/em\u003e (J.E. Smith) (Lepidoptera: Noctuidae), commonly known as the fall armyworm (FAW), is a notorious polyphagous insect indigenous to the Western Hemisphere's warmer climates (Kumar et al. \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). The pest is reported to feed on a remarkably wide variety of hosts, with records indicating over 350 plant species within 76 families (Montezano et al. \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2018\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe urgent need for biological control of the fall armyworm arises from its devastating impact on crop yields and the severe limitations of sole reliance on chemical insecticides, which include rising resistance, environmental harm, and high costs for smallholder farmers (Ghafar et al. \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). Host location is a central process in the life of parasitoid because finding suitable hosts is key for its reproduction success (Turlings and Erb \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). At the beginning of foraging, parasitoids use different strategies and cues for locating their hosts, depending on the host stage. For parasitoids of eggs, the host location process may imply a higher challenge in comparison to larval or adult parasitoids, considering that eggs do not feed nor defecate, and thus do not emit long- range volatiles that can be used as cues by their natural enemies (Ngumbi et al. 2021).\u003c/p\u003e \u003cp\u003eSeveral studies have investigated the role of herbivore-induced volatiles during the host searching by insect parasitoids using the tritrophic model of maize-Lepidopteran-parasitoids (Ortiz-Carreon et al. \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Plants emit volatile compounds that have a major impact on their environment (Tholl et al. 2018), and when undergoing herbivory, they produce specific Herbivore-Induced Plant Volatiles (HIPVs). These HIPVs function as indirect defense signals, providing olfactory cues that are critical for hymenoptera parasitoids to locate their hosts (Turlings et al. 1998). The induced changes in the plant's volatile blend convey information about the attacking herbivore and attract its natural enemies, thereby forming a key component of the plant's indirect defense response (Sharma et al. \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2017\u003c/span\u003e).These volatiles act as signals to distal tissues of the plant itself, as well as to other organisms in the environment, providing information on the identity, location, and health of the plant (Karalija et al. \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe most common strategy used by egg parasitoids to find their hosts is exploiting chemical cues related to adult hosts, including sex, antisex, and aggregation pheromones, or host adult residues, such as genital secretions and scales that line the egg masses. A second strategy is to use of plant volatiles, commonly constitutive volatiles that are used as long- range chemical cues by egg parasitoids, though herbivory- induced plant volatiles may be more attractive cues for parasitoids by healthy or mechanically-damaged plants (Tepa-Yotto et al. \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). The most common strategy used by egg parasitoids to find their hosts is exploiting chemical cues related to adult hosts, including sex, antisex, and aggregation pheromones, or host adult residues, such as genital secretions and scales that line the egg masses.\u003c/p\u003e"},{"header":"METHODS AND MATERIALS","content":"\u003cp\u003e \u003cb\u003eFAW Culture\u003c/b\u003e The \u003cem\u003eS. frugiperda\u003c/em\u003e larvae were collected from fall armyworm rearing laboratory, Department of Entomology, Rajasthan College of Agriculture, MPUAT, Udaipur. Larvae were maintained individually to avoid cannibalism. Later, larvae were taken to the insectary to be reared using an artificial diet under controlled conditions of temperature (26\u0026thinsp;\u0026plusmn;\u0026thinsp;2\u0026deg;C), relative humidity (75\u0026thinsp;\u0026plusmn;\u0026thinsp;5%), and a photoperiod of 12:12 h (light/dark) (Kumar et al. \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). The pupae were kept in glass cages until adult emergence. Ten adults (5 females and 5 males) were place in Kraft paper bags for mating and oviposition. Adults were feed a 10% honey solution dispensed on cotton wool, and was kept under the conditions described above. \u003cem\u003eS. frugiperda\u003c/em\u003e egg masses deposited in Kraft paper bags were collected daily.\u003c/p\u003e \u003cp\u003e \u003cb\u003eParasitoids Culture\u003c/b\u003e The \u003cem\u003eS. frugiperda\u003c/em\u003e larvae parasitized by parasitoid/s (\u003cem\u003eTelenomus remus\u003c/em\u003e and \u003cem\u003eTrichogramma chilonis\u003c/em\u003e) were collected from biological control laboratory, Department of Entomology, Rajasthan College of Agriculture, MPUAT, Udaipur and used as rearing stock for culturing wasps for bioassays. Three days-old adult parasitoids were placed in Petri dishes in a 1:1 sex ratio for mating. After mating, an egg mass of \u003cem\u003eS. frugiperda\u003c/em\u003e offered to each \u003cem\u003eTelenomus remus\u003c/em\u003e and \u003cem\u003eTrichogramma chilonis\u003c/em\u003e female for oviposition. The egg masses were removed 24 h later, or until FAW larvae hatched; egg masses or neonate larvae were placed into containers, with artificial diet. Ten days later, \u003cem\u003eS. frugiperda\u003c/em\u003e larvae were individually placed into transparent box until the emergence of adult of \u003cem\u003eTelenomus remus\u003c/em\u003e and \u003cem\u003eTrichogramma chilonis\u003c/em\u003e parasitoids. The parasitoids were fed with honey drops placed in the small container and kept under the laboratory conditions described above.\u003c/p\u003e \u003cp\u003e \u003cb\u003eKairomones Collection from egg Surface\u003c/b\u003e To eliminate natural kairomones from the egg surface, 10 egg masses (0\u0026ndash;24 hr old; about 2,500 eggs) taken from culture were washed twice in hexane (10 ml). The hexane egg wash rinsate was filtered using WhatmanTM No. 1 filter paper. The filtrates were kept at -20\u0026deg;C and utilized as stock for further diluting.\u003c/p\u003e \u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eFAW female moth scale collection and extraction\u003c/h2\u003e \u003cp\u003eFemale fall armyworm scales were taken from freshly emerged (0\u0026ndash;24 hrs old) laboratory-reared cultures by immobilising them at 0\u0026ndash;2\u0026deg;C. The wings were removed from around 30 moths. The wing scales were extracted by shaking vigorously in a chilled shaker for 2 hours in 200 ml of analytical grade hexane and filtered through Whatman No.1 filter paper. The filtrate was kept at -20\u0026deg;C for Gas chromatography linked to mass spectrometry.\u003c/p\u003e \u003cp\u003e \u003cb\u003eGas Chromatography Coupled with Mass Spectrophotometry (GC-MS)\u003c/b\u003e Using a GC-MS Agilent 7890B GC system with Mass Spectrometry (Agilent 5977 MSD), the chemical compositions of eggs and scales kairmone cues were examined. This was done in accordance with the method previously outlined by Jayanthi et al. (\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2021\u003c/span\u003e), with a few minor adjustments. The samples were examined using an Agilent J \u0026amp; W (HP-5 MS UI) capillary column that was 30 m long, 0. 250 mm in diameter, and had a film thickness of 0.25 \u0026micro;m. Helium was used as the carrier gas, and the thermal program was configured as previously mentioned. The flow rate was 1 mL/min. AMU varied between 40 and 450, while MS was operating in full scan mode (70 eV). 1.0 microlitre of the sample was inserted in splitless mode ratio (40 mL/min) with the injection temperature of 270\u0026ordm;C. The individual chemicals were initially identified by comparing GC retention time, Kovats index (C7 to C30 homologous series of n-alkenes as standard, Sigma-Aldrich; Kovats 1965) and comparing the mass spectra with spectral library NIST 14. The identified compounds were further authenticated and quantified by using a single point external standard quantification method by using authentic samples of standards (Ciotlaus et al. 2024) and the compounds for which synthetic standards are unavailable were identified tentatively based on NIST 14 spectral library.\u003c/p\u003e \u003cp\u003e \u003cb\u003eBioassays\u003c/b\u003e To evaluate the behavioral responses of naive parasitoid to maize plants with and without eggs of fall armyworm, we were evaluated the responses of the parasitoids to maize plants with and without FAW eggs. Three seedlings were placed inside a collapsible mesh cage with 10 \u003cem\u003eS. frugiperda\u003c/em\u003e gravid females 5\u0026ndash;8 days old during one night to obtain plants with eggs. Moths were supplied with sugar water on cotton wool. The seedlings with 2\u0026ndash;4 egg masses (oviposition-treated seedlings) were selected for the bioassays. For plants without eggs, seedlings of the same age as the oviposition treated seedlings but without FAW oviposition were used (egg-free seedlings). The following comparisons were performed: (1) Egg-free seedlings versus clean air, (2) Oviposition-treated seedlings versus clean air, (3) Egg-free seedlings plus 3\u0026ndash;4 FAW egg masses oviposited on Kraft paper versus clean air, (4) Egg-free seedlings versus oviposition-treated seedlings (5) Egg-free seedlings plus 3\u0026ndash;4 FAW egg masses oviposited on Kraft paper versus egg-free seedlings and (6) Egg-free seedlings plus 3\u0026ndash;4 FAW egg masses oviposited on Kraft paper versus Kraft paper. The Kraft paper with the egg masses was placed on the seedlings leaves. Seedlings received eggs 12\u0026ndash;14 h before the evaluation, wherase, behavioral responses of experienced parasitoid to maize plants with and without eggs of FAW, we were investigated that whether oviposition experience influences the responses of parasitoid females to maize plants with and without eggs. Parasitoid females with oviposition experience were obtained single females on seedlings previously oviposited by \u003cem\u003eS. frugiperda\u003c/em\u003e so that they have contact with the plant and detect and parasitize eggs. The parasitoid females were removed 2 min later. The experienced females were tested the next day in the Y-tube olfactometer. The following comparisons were performed: (1) egg-free seedlings versus oviposition-treated seedlings, and (2) oviposition-treated seedlings versus 3\u0026ndash;4 egg masses oviposited on Kraft paper. The bioassays were performed following the methodology described in the above section. While to test the behavioral responses of Naive parasitoid to egg masses of \u003cem\u003eS. frugiperda\u003c/em\u003e, was evaluated that whether naive parasitoid females are attracted to host egg masses. Because Kraft paper was used as an oviposition substrate for FAW rearing; this material was included as a treatment in this experiment. The following comparisons were performed: (1) Kraft paper versus clean air, (2) 3\u0026ndash;4 egg masses oviposited on Kraft paper versus clean air and (3) 3\u0026ndash;4 egg masses oviposited on Kraft paper versus Kraft paper (Roque-Romero et al. \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2020\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e"},{"header":"RESULT","content":"\u003cp\u003e \u003cb\u003eIdentification of Chemical Cues from Egg Surface Extracts\u003c/b\u003e The surface of eggs of FAW volatile emmisions were worked out using Gas Chromatography. The eggs surface emmited a total of 24 volatiles from 11 functional groups including sulfure compounds aldehydes, terpenes, esters, halogentad compounds, alcohols/ ethers, ketones, heterocytes, thioether, hydrocarborn, lignans and siloxanes, compounds have been identified. GCMS anaysis revealed siginificant differences among the functional chemical groups or classes of cues in surface of egg of \u003cem\u003eS\u003c/em\u003e. \u003cem\u003efrugiperda\u003c/em\u003e (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eAmong the chemical groups esters (Oxalic acid, cyclohexyl propyl ester, Oxalic acid, cyclohexyl isobutyl ester, Methyl allyl diglycolcarbonate, Carbonic acid, monoamide, N-pentyl-, decyl ester and Benzoic acid, 3,3'-[1,3-phenylenebis(carbonylimino)] bis-, dimethyl) were prodominent followed by hetereocycles (N-(2-Propynyl)aziridine, Thiophen-2-methylamine, N-(2-fluorophenyl)-, 2-(p-(Dimethylamino) phenyl) benzimidazole, 4H-1,2,4-Triazole, 4-ethyl- and 3-Pyridin-4-yl-2-(pyridin-4-yl-p-tolylaminomethyl)-3-p-tolylamino-propionitrile.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003e\u003cb\u003eGC-MS profile of chemical cues of eggs of\u003c/b\u003e \u003cb\u003eSpodoptera frugiperda\u003c/b\u003e\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"3\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eChemical Group\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSpecific Compounds\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eArea percentage\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e1. Esters\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eOxalic acid, cyclohexyl propyl ester\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e24.36\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eOxalic acid, cyclohexyl isobutyl ester\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2.14\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMethyl allyl diglycolcarbonate\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.01\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCarbonic acid, monoamide, N-pentyl-, decyl ester\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e13.50\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBenzoic acid, 3,3'-[1,3-phenylenebis(carbonylimino)]bis-, dimethyl ester\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.009\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e2. Aldehydes\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePentanal, 2,2-dimethyl-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e19.66\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e3. Ketones/Epoxides\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eEthanone, 1-(3-methyloxiranyl)-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e15.42\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eBromoacetone\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.28\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e4. Ethers\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e4-Methyl-2-pentanol, methyl ether\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1.08\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3-Methoxy-3-methylbutanol\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.01\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e5. Thioethers\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eButanoic acid, 3-(methylthio)-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e4.60\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e6. Halogenated Compounds\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1,3-Butadiyne, 1,4-difluoro-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1.48\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eHexanee, 2-bromo-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e3.44\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1(2H)-naphthalenone, 2-bromo-3,4-dihydro-6-methoxy-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2.54\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e7. Heterocycles/Nitrogenous\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eN-(2-Propynyl)aziridine\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.59\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eThiophen-2-methylamine, N-(2-fluorophenyl)-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1.69\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e2-(p-(Dimethylamino)phenyl)benzimidazole\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.02\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e4H-1,2,4-Triazole, 4-ethyl-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1.79\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e3-Pyridin-4-yl-2-(pyridin-4-yl-p-tolylaminomethyl)-3-p-tolylamino-propionitrile\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.12\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e8. Hydrocarbons\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eHepta-1,2,6-heptatriene\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.54\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eButane, 2,2-dimethyl-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.10\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e9. Cyclic Carbonates\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1,3-Dioxol-2-one\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e1.57\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e10. Siloxanes\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCyclotetrasiloxane, octamethyl-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.02\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e11. Lignans\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSchizandrin\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.004\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003eIdentification of chemical compounds presents in headspace volatiles of wings cues of\u003c/b\u003e \u003cb\u003eS. frugiperda\u003c/b\u003e A total of 22 volatiles from 10 functional groups, including esters, aldehydes, ketones,ethers,heterocycles, halogenated compounds, nitrogenous compounds,siloxanes, aromaticsand fluorinated compoundshave been identified from the surfaceof FAW wings(Table\u0026nbsp;4.39 and Fig.\u0026nbsp;4.64). The esters were the dominant group in the wings of \u003cem\u003eS. frugiperda\u003c/em\u003e with five compounds, including Oxalic acid, cyclohexyl propyl ester, Oxalic acid, cyclohexyl isobutyl ester, Methyl allyl diglycolcarbonate, Carbonic acid, monoamide, N-pentyl-, decyl ester and Benzoic acid, 3,3'-[1,3-phenylenebis(carbonylimino)]bis-, dimethyl ester (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003e\u003cb\u003eGC-MS profile of chemical cues of wings of\u003c/b\u003e \u003cb\u003eSpodoptera frugiperda\u003c/b\u003e\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"3\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eChemical Group\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eArea percentage\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eCompound Name\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e1. Esters\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e36.73\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eOxalic acid, cyclohexyl propyl ester\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e4.27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eOxalic acid, cyclohexyl isobutyl ester\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0.014\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eOxalic acid, dineopentyl ester\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAcetyl valeryl\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0.04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eSulfurous acid, 2-ethylhexyl hexyl ester\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e1.14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eTerephthalic acid, isobutyl 2,2,3,3,3-pentafluoropropyl ester\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e2. Aldehydes\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e30.67\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePentanal, 2,2-dimethyl-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e3. Ketones/Epoxides\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e6.35\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eEthanone, 1-(3-ethyloxiranyl)-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0.62\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eAcetyl bromide\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e4. Ethers\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e6.35\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eEthane, 1,1-dimethoxy-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eButane, 1-(1-methylpropoxy)-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e5. Heterocycles\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0.12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eOxazole\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0.56\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e(S)-(-)-1-Amino-2-(methoxymethyl)-pyrrolidine\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0.11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eN-[4-(1H-Benzoimidazol-2-yl)-phenyl]-acetamide\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e6. Halogenated Compounds\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0.86\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1-Buten-3-yne, 1-chloro-, (E)-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0.61\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eCyclopropane, 2-bromo-1,1,3-trimethyl-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e7. Nitrogenous Compounds\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0.09\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eCyclohexanee, nitro-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e8. Siloxanes\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e8.57\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eSilicic acid, diethyl bis(trimethylsilyl) ester\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e9. Aromatics\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0.52\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3,4-Dimethoxy-N-[2-(2-methoxyphenoxy)ethyl]benzamide\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003e10. Fluorinated Compounds\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e0.06\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eBisphenol, O,O'-bis(pentafluoropropionyl)-\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003eBehavioural response of na\u0026iuml;ve female of\u003c/b\u003e \u003cb\u003eTelenomus remus\u003c/b\u003e \u003cb\u003eto maize plants with and without eggs of\u003c/b\u003e \u003cb\u003eS. frugiperda\u003c/b\u003e \u003cb\u003ein Y- olfactometer during single- choice bioassay\u003c/b\u003e The findings of present study revlealed that the responses of na\u0026iuml;ve female wasps of \u003cem\u003eT. remus\u003c/em\u003e to odors from eggs free seedilings were significantly [χ\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;12.175; \u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.0001], Ovipostion treated seedlings [χ\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;15.000; \u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.0001], egg-free seedlings plus 3\u0026ndash;4 FAW egg masses oviposited on Kraft paper [χ\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;10.756; \u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.001] over clean air. The Oviposition treated seedling odour also significntly attrarctive to na\u0026iuml;ve female wasps [χ\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;20.000; \u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.0001] over Egg- free seedlings. Egg-free seedlings plus 3\u0026ndash;4 FAW egg masses oviposited on Kraft paper [χ\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;14.118; \u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.0001] and Egg-free seedlings plus 3\u0026ndash;4 FAW egg masses oviposited on Kraft paper [χ\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;12.593] significantly attracted over kraft paper. Present study indicated that the odors released from oviposited plants more attractive than normal plant and plant odour is attractive to parasitoids than clean air or other odors (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cb\u003eBehavioural response of experienced female of\u003c/b\u003e \u003cb\u003eTelenomus remus\u003c/b\u003e \u003cb\u003eto maize plants with and without eggs of\u003c/b\u003e \u003cb\u003eS. frugiperda\u003c/b\u003e \u003cb\u003ein Y- olfactometer during single- choice bioassay\u003c/b\u003e The experienced female \u003cem\u003eT.remus\u003c/em\u003e were more attracted to odors from oviposited seedlings when compared to odors from egg-free seedlings [\u003cb\u003eχ\u003c/b\u003e\u003csup\u003e\u003cb\u003e2\u003c/b\u003e\u003c/sup\u003e\u0026thinsp;\u003cb\u003e=\u003c/b\u003e\u0026thinsp;20.000; \u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.0001] and Ovipostion treated seedlings [\u003cb\u003eχ\u003c/b\u003e\u003csup\u003e\u003cb\u003e2\u003c/b\u003e\u003c/sup\u003e\u0026thinsp;\u003cb\u003e=\u003c/b\u003e\u0026thinsp;12.926; \u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.0001] atrracted more females over 3\u0026ndash;4 FAW egg masses oviposited on Kraft paper. Results of this experiment reveals that both plant volatiles and egg cues more attractive together (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cb\u003eBehavioural response of na\u0026iuml;ve female of\u003c/b\u003e \u003cb\u003eTelenomus remus\u003c/b\u003e \u003cb\u003eto egg masses of\u003c/b\u003e \u003cb\u003eS. frugiperda\u003c/b\u003e \u003cb\u003ein Y- olfactometer during single- choice bioassay\u003c/b\u003e The na\u0026iuml;ve females of \u003cem\u003eT. remus\u003c/em\u003e had not show a significant preference for odors from Kraft paper and clean air [\u003cb\u003eχ\u003c/b\u003e\u003csup\u003e\u003cb\u003e2\u003c/b\u003e\u003c/sup\u003e\u0026thinsp;\u003cb\u003e=\u003c/b\u003e\u0026thinsp;12.175; \u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.005]. However, the \u003cem\u003eT. remus\u003c/em\u003e females were attracted to odors emmited from egg masses oviposited in Kraft paper when compared to the clean air and kraft paper [\u003cb\u003eχ\u003c/b\u003e\u003csup\u003e\u003cb\u003e2\u003c/b\u003e\u003c/sup\u003e\u0026thinsp;\u003cb\u003e=\u003c/b\u003e\u0026thinsp;15.000; \u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.0001 and \u003cb\u003eχ\u003c/b\u003e\u003csup\u003e\u003cb\u003e2\u003c/b\u003e\u003c/sup\u003e\u0026thinsp;\u003cb\u003e=\u003c/b\u003e\u0026thinsp;10.756; \u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.001, respectively]. The above mentioned findings are clearly evident that volatiles and cues released by plant and egg surface respectively are attaracted to egg parasitoids (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cb\u003eBehavioural response of na\u0026iuml;ve female of\u003c/b\u003e \u003cb\u003eTrichogrmma chilonis\u003c/b\u003e \u003cb\u003eto maize plants with and without eggs of\u003c/b\u003e \u003cb\u003eS. frugiperda\u003c/b\u003e \u003cb\u003ein Y- olfactometer during single- choice bioassay\u003c/b\u003e The findings of present study revlealed that the responses of na\u0026iuml;ve female wasps of \u003cem\u003eT. chilonis\u003c/em\u003e to odors from eggs free seedilings were significantly [χ\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;15.556; \u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.0001], Ovipostion treated seedlings [χ\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;6.667; \u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.0001], egg-free seedlings plus 3\u0026ndash;4 FAW egg masses oviposited on Kraft paper [χ\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;20.000; \u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.001] over clean air. The Oviposition treated seedling odour also significntly attrarctive to na\u0026iuml;ve female wasps [χ\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;12.926; \u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.0001] over Egg- free seedlings. Egg-free seedlings plus 3\u0026ndash;4 FAW egg masses oviposited on Kraft paper [χ\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;13.003; \u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.0001] and Egg-free seedlings plus 3\u0026ndash;4 FAW egg masses oviposited on Kraft paper [χ\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;20.000; \u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.0001] significantly attracted over kraft paper. Present study indicated that the odors released from oviposited plants more attractive than normal plant and plant odour is attractive to parasitoids than clean air or other odors (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cb\u003eBehavioural response of experienced female of\u003c/b\u003e \u003cb\u003eTrichogrmma chilonis\u003c/b\u003e \u003cb\u003eto maize plants with and without eggs of\u003c/b\u003e \u003cb\u003eS. frugiperda\u003c/b\u003e \u003cb\u003ein Y- olfactometer during single- choice bioassay\u003c/b\u003e The experienced female of \u003cem\u003eT. chilonis\u003c/em\u003e were more attracted to odors from oviposited seedlings when compared to odors from egg-free seedlings [\u003cb\u003eχ\u003c/b\u003e\u003csup\u003e\u003cb\u003e2\u003c/b\u003e\u003c/sup\u003e\u0026thinsp;\u003cb\u003e=\u003c/b\u003e\u0026thinsp;19.000; \u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.0001] and Ovipostion treated seedlings [\u003cb\u003eχ\u003c/b\u003e\u003csup\u003e\u003cb\u003e2\u003c/b\u003e\u003c/sup\u003e\u0026thinsp;\u003cb\u003e=\u003c/b\u003e\u0026thinsp;20.000; \u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.0001] atrracted more females over 3\u0026ndash;4 FAW egg masses oviposited on Kraft paper. Results of this experiment reveals that both plant volatiles and egg cues more attractive together (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cb\u003eBehavioural response of na\u0026iuml;ve female of\u003c/b\u003e \u003cb\u003eTrichogrmma chilonis\u003c/b\u003e \u003cb\u003eto egg masses of\u003c/b\u003e \u003cb\u003eS. frugiperda\u003c/b\u003e \u003cb\u003ein Y- olfactometer during single- choice bioassay\u003c/b\u003e The na\u0026iuml;ve females of \u003cem\u003eT. chilonis\u003c/em\u003e had not show a significant preference for odors from Kraft paper and clean air [\u003cb\u003eχ\u003c/b\u003e\u003csup\u003e\u003cb\u003e2\u003c/b\u003e\u003c/sup\u003e\u0026thinsp;\u003cb\u003e=\u003c/b\u003e\u0026thinsp;16.296; \u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.0001]. However, the \u003cem\u003eT. remus\u003c/em\u003e females were attracted to odors emmited from egg masses oviposited in Kraft paper when compared to the clean air and kraft paper [\u003cb\u003eχ\u003c/b\u003e\u003csup\u003e\u003cb\u003e2\u003c/b\u003e\u003c/sup\u003e\u0026thinsp;\u003cb\u003e=\u003c/b\u003e\u0026thinsp;19.000; \u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.0001 and \u003cb\u003eχ\u003c/b\u003e\u003csup\u003e\u003cb\u003e2\u003c/b\u003e\u003c/sup\u003e\u0026thinsp;\u003cb\u003e=\u003c/b\u003e\u0026thinsp;20.000; \u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.001, respectively]. The above mentioned findings are clearly evident that volatiles and cues released by plant and egg surface respectively are attaracted to egg parasitoids (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cb\u003eBehavioural response of naive female of\u003c/b\u003e \u003cb\u003eTelenomus remus\u003c/b\u003e \u003cb\u003eto wing and egg cues extraction of\u003c/b\u003e \u003cb\u003eS. frugiperda\u003c/b\u003e \u003cb\u003ein Y- olfactometer during single- choice bioassay\u003c/b\u003e The behavioural response of naive female of \u003cem\u003eTelenomus remus\u003c/em\u003e to wing and egg cues extraction of \u003cem\u003eS. frugiperda\u003c/em\u003e in Y- olfactometer during single- choice bioassay. The egg and wing cues of \u003cem\u003eS. frugiperda\u003c/em\u003e were signficantly more attractive when compared with hexan solvent [χ\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;3.951; \u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.0001 and χ\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;20.000; \u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.0001, respectively]. The females of \u003cem\u003eT. remus\u003c/em\u003e were also significantly attracted to eggs cues than wing cues [χ\u003csup\u003e2\u003c/sup\u003e\u0026thinsp;=\u0026thinsp;9.474; \u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.0001] (Fig.\u0026nbsp;\u003cspan refid=\"Fig7\" class=\"InternalRef\"\u003e7\u003c/span\u003e).\u003c/p\u003e"},{"header":"DISCUSSION","content":"\u003cp\u003eDuring the present investigation we were found that the experienced female \u003cem\u003eT. remus\u003c/em\u003e were more attracted to odours from oviposited seedlings and Ovipostion treated seedlings than egg masses oviposited on Kraft paper. The study also found that both plant volatiles and egg cues are more attractive together, indicating that both plant and egg surface odours are attracted to egg parasitoids. The present study also reveals that na\u0026iuml;ve female \u003cem\u003eTrichogrmma chilonis\u003c/em\u003e wasps were significantly attracted to odours from egg-free seedlings, Ovipostion-treated seedlings, and egg-free seedlings plus 3\u0026ndash;4 FAW egg masses oviposited on Kraft paper. The odours released from oviposited plants were more attractive than normal plants and plant odour was more attractive to parasitoids than clean air or other odours. Experienced female \u003cem\u003eT. chilonis\u003c/em\u003e were more attracted to odours from oviposited seedlings and Oviposition-treated seedlings. The study also found that both plant volatiles and egg cues are more attractive together, with \u003cem\u003eT. remus\u003c/em\u003e females being attracted to odours emitted from egg masses oviposited on Kraft paper. Present findings are closely associated with those of Roque-Romero et al. (\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) who reported that the \u003cem\u003eC.insularis\u003c/em\u003e females responded to volatiles released by host egg masses, females (sex pheromones), and maize seedlings with or without FAW eggs. Females can distinguish between maize seedlings with and without FAW eggs due to the parasitoids' enhanced reactivity to host and maize seedling volatiles following their initial oviposition event. The authors presented theories to explain their findings, including the idea that FAW females limit herbivore-induced plant volatiles to avoid attracting parasitoids and predators. Some plant species change their volatile profile after herbivore oviposition, attracting parasitoids and predators from a distance (Nascimento et al. 2023). These chemical signals help the natural enemies of the herbivores locate infested plants, thus providing the plants with an indirect defense mechanism (Wang et al. \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). This has been demonstrated in numerous studies showing that parasitoid wasps are drawn to the volatiles produced by plants under attack from their host insects (Nideesh et al. 2025). Similarly, the study found that \u003cem\u003eC. insularis\u003c/em\u003e females are attracted to maize volatiles induced by larval feeding, but they are not the host stages of FAW larvae. The study confirmed that na\u0026iuml;ve \u003cem\u003eC. insularis\u003c/em\u003e are attracted to FAW egg mass volatiles and female sex pheromone, as well as healthy maize seedling volatiles. The innate response to egg masses is not significant when the plant is present, or the parasitoid attraction increases with plant volatiles. Experienced female parasitoids can discriminate between previously oviposited maize seedlings and egg-free ones (Ortiz-Carreon et al. \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). The volatiles obtained from larval and adult stages of \u003cem\u003ePhthorimaea operculella\u003c/em\u003e, adult cues were observed to be more efficient in the alteration of foraging behavior of the targeted parasitoid, \u003cem\u003eC. blackburni\u003c/em\u003e and the present study revealed that the \u003cem\u003eC. blackburni\u003c/em\u003e preferred cues could be exploited to enhance the host searching efficiency during IPM releases. Several parasitoids are known to exploit these plant-provided cues to locate their hosts. One such parasitoid is the generalist \u003cem\u003eCotesia marginiventris\u003c/em\u003e, which is, among others, attracted to maize volatiles induced by caterpillar damage (Singh et al. \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). This has been demonstrated in numerous studies showing that parasitoid wasps are drawn to the volatiles produced by plants under attack from their host insects (Nideesh \u003cem\u003eet al.\u003c/em\u003e 2025). Similarly, \u003cem\u003eOoencyrtus\u003c/em\u003e spp. were found in plots with both \u003cem\u003eA. contorta\u003c/em\u003e and \u003cem\u003eS. montela\u003c/em\u003e larvae, indicating that the VOCs produced by feeding damage and larval saliva attract parasitoids (Davidson Lowe and Ali 2021; Gebreziher \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Moujahed et al. \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2014\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eIn conclusion, we evaluated that both plant volatiles and egg cues are more attractive together, indicating that both plant and egg surface odours are attracted to egg parasitoids. The present study also reveals that na\u0026iuml;ve female \u003cem\u003eTrichogrmma chilonis\u003c/em\u003e wasps were significantly attracted to odours from egg-free seedlings, Ovipostion-treated seedlings, and egg-free seedlings plus 3\u0026ndash;4 FAW egg masses oviposited on Kraft paper. The odours released from oviposited plants were more attractive than normal plants and plant odour was more attractive to parasitoids than clean air or other odours.\u003c/p\u003e"},{"header":"Declarations","content":"\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eThe writing and technical corrections done by Devendra Jain and Heenashree Mansion. An experiment conducted by Amit Kumar and S. Ramesh babu\u003c/p\u003e\u003ch2\u003eAcknowledgments\u003c/h2\u003e \u003cp\u003eWe are thankful Department of Entomology, Rajasthan College of Agriculture, Maharana Pratap University of Agriculture and Technology, Udaipur, Rajasthan (India) for technical assistance. The writing and technical correction done by \u0026hellip;\u0026hellip;\u0026hellip;..\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eCiotlăuș I, Balea A, Pojar-Feneșan M, Filip MR, Vlassa M (2024) GC-MS And Hplc Chromatographic Profile Of Majority Volatile And Phenolic Compounds Of Some Medicinal Plants From Romania. Studia Universitatis Babes-Bolyai, \u003cem\u003eChem\u003c/em\u003e, 69\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDavidson-Lowe E, JG A (2021) Herbivore-Induced Plant Volatiles Mediate Behavioral Interactions Between a Leaf-Chewing and a Phloem-Feeding Herbivore. Bas App Eco 53:39\u0026ndash;48\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGebreziher HG (2020) Advances in Herbivore-Induced Plant Volatiles (HIPVs) as Plant Defense and Application Potential for Crop Protection. 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Pest Manage Scie 81:3674\u0026ndash;3684\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Telenomus remus, Trichogramma chilonis, Egg cues, Fall armyworm, Behavioral bioassays","lastPublishedDoi":"10.21203/rs.3.rs-8768024/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8768024/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThe Fall Armyworm is a highly polyphagous, dreaded insect pest belonging to the order lepidoptera. During this study we, evaluated in behavioral bioassays, female \u003cem\u003eSpodoptera frugiperda\u003c/em\u003e moths and two egg parasitoid species were tested for their responses to maize plant volatiles. Based on the combined findings, both \u003cem\u003eTrichogramma chilonis\u003c/em\u003e and \u003cem\u003eTelenomus remus\u003c/em\u003e egg parasitoids rely on combined plant and host egg cues for optimal attraction. Experienced females of both species show a stronger preference for odours from oviposited plants over plants or egg cues alone.\u003c/p\u003e","manuscriptTitle":"Behavioral Responses of Egg Parasitoids Telenomus remus and Trichogramma chilonis to Kairomonal cues released from Eggs and Wings of Fall Armyworm (Spodoptera frugiperda J.E. Smith)","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-02-12 18:44:44","doi":"10.21203/rs.3.rs-8768024/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"8340a583-112a-48ea-9317-431718c21f23","owner":[],"postedDate":"February 12th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2026-02-24T15:40:57+00:00","versionOfRecord":[],"versionCreatedAt":"2026-02-12 18:44:44","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8768024","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8768024","identity":"rs-8768024","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}