Effectiveness of jasmonate stimulation on attraction of Cotesia marginiventris via olfactory cues on BT- and non-BT-cotton plants

Effectiveness of jasmonate stimulation on attraction of Cotesia marginiventris via olfactory cues on BT- and non-BT-cotton plants | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Effectiveness of jasmonate stimulation on attraction of Cotesia marginiventris via olfactory cues on BT- and non-BT-cotton plants NABIL EL-WAKEIL, NAWAL GAAFAR, SAAD ALKAHTANI This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5306734/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 It is well documented that plants damaged with insect species can attract specific biocontrol agents throughout the emission of insect-induced volatiles. A four-armed olfactometer was used to inspect olfactory reactions of Cotesia marginiventris . Behavioural tests for studying parasitoid attraction to specific odours are assessed. Flying tunnel investigations could be evaluated with three odour sources plus control plants at a time. The attractiveness of female parasitoid C. marginiventris was tested by applying several treatments: jasmonate induced plants and plants fed by Spodoptera exigua on Bacillus thuringiensis (BT)(genetic modified GM) and non- GM cotton plants. Three criteria were compared and studied. Search time and choose time and visit numbers for each zone. The obtained outcomes showed that the plants induced with jasmonate were more attractive than those plants fed by two larvae, while those plants fed by one larva were the least attractive. Whereas, it was found that females of C. marginiventris significantly chose more jasmonate feeding areas than larval feeding areas on GM cotton plants, and the equivalent effect was detected on non-GM cotton plants. Whereas C. marginiventris females spent more time in the jasmonate areas than the rest of the treatments; While the time spent on transgenic cotton plants was longer than the time spent on non-transgenic cotton plants. Whereas in the non-Jasmonate treatments, females spent more time using the GM cotton than the non-GM cotton, the same trend of effects had been detected in the choice of time and visit frequency for that treatment. Jasmonate stimulation of volatile substances is a good way to increase the attractiveness of parasitoids to their insect hosts to maximize the efficacy of the role of biological control agents. It is proposed that a genetic modification approach to modify the volatiles in these plants could enhance the attractiveness and increase the role assigned to natural enemies. This procedure will cause a more drastic change in the volatiles of those plants. This scientific direction is a complementary role, with transgenic resistance, which will lead to selection of inducible promoters with tissue-specific expression. Cotesia marginiventris Olfactometer Jasmonate Cotton Beet armyworm Figures Figure 1 Figure 2 Figure 3 Figure 4 1. INTRODUCTION The transgenic cotton plants produce a type of BT toxin that helps providing high levels of resistance to insect groups such as Lepidoptera and Coleoptera that feed on important economic crops, and may have side effect on natural enemies (Schuler 1999 a,b; El-Wakeil et al. 2003). The expected effect of these genetic modified plant groups handled with jasmonate on biocontrol agents in general and on parasitoid wasps in particular, has not yet acknowledged the sufficient attention (Gu and Dorn 2000; Turlings et al. 2005). There is a gab to understand effectiveness of Bt cotton with effect of Jasmonic acid and their induced volatiles for attracting the biological control agents. In the case of plant volatiles, little is known about their beneficial biological roles for plants. While it is clear that their function is not limited to attracting parasitoids or predators (Mumm and Dicke 2010; Clavijo McCormick et al. 2012); there are key components that are common across plant species. Single compounds may attract natural enemies to foliage damaged by herbivores. (Bruce et al. 2005; Clavijo McCormick et al. 2012; Ali et al. 2022, 2023). In another case, the mixture of small compounds might tell several parasitoids where their host insect pests are located and help killing them biologically (Clavijo McCormick et al. 2014; Ali et al. 2021a). It is scientifically proven that plants produce jasmonic acid after exposure to insect infestations, which leads to increased production of some compounds that participate in resistance against insect pests (de Boer et al. 2008). Jasmonic acid has also been known to be involved in the production of plants to release volatile compounds; insect pests and applying of jasmonic acid to leaves trigger volatile terpenoids that have been specifically linked to attracting natural enemies (Dicke et al. 1999; Mumm and Dicke 2010; Liu et al. 2017). Jasmonic acid applications on plants produce proteinase inhibitors and polyphenol oxidases, which helps reducing the number of insect pests and increasing number of biological control agents in the field. (Thaler 1999; D'Alessandro and Turlings 2005, 2006; Pinto-Zevallos et al. 2016 Ali et al. 2021b). Insect feeding on the plant helps to modify the levels of volatile substances by producing chemical combinations associated with damage to plant tissues, which increases the induced plant defenses (Selig et al. 2016). These volatile substances increase the attraction of the natural enemies of these insect pests (Kessler and Baldwin 2001; Pickett and Glinwood 2007; Pinto-Zevallos et al. 2016; Turlings and Erb 2018; Xiu et al. 2019). The volatiles apparently released constitutively by plants might attract many natural enemy species (Loughrin et al. 1995; Halitschke et al. 2000; Snoeren et al. 2010; Takemoto and Takabayashi 2015; Piesik et al. 2022). The plant's release of chemical compounds that help stimulate the attraction of natural enemies to insects, therefore, this mechanism is considered an indirect means of defense against plants potentially infested by insects and may lead to insect resistance (Price et al. 1980; Mumm and Dicke 2010; Liu et al. 2017), It is known that insect repellent chemical compounds are a direct mechanism for plant defense, or in other words plant resistance to antitoxins against other insect pests. (Dicke et al. 1990 a,b; Wei et al. 2011; Schuman et al. 2012). Cotesia marginiventris (Cresson) it is an endo larval parasitoid that parasitizes many Lepidopteran insect pests. It is certain that this parasitoid responds to contact chiromones present in the host's by-products like silk and saliva. It also responds strongly to the host's feces, as well as the damage to the plant host's feeding caused by insect larvae (Turlings et al. 1990, 1991; Ngumbi et al. 2005; Pålsson et al. 2021). A volatile or called volicitin; it is a key source of natural enemy's attraction to infested plants (17-hydroxlinolenoyl-l-Gln) (Liu et al. 2017). This volatile substance (volicitin) was separated, identified, and attempted to be created from the oral secretions of the beet armyworm Spodoptera exigua (Turlings et al. 2000; El-Wakeil 2003). C. marginiventris , it is hypothesized that this parasitoid responds normally to volatile substances found in some plants, particularly if fed by S. exigua larvae. During some foraging experiments, females of these parasitoids seem to learn how to respond to a combination of volatiles and odors specific to a plant compound of a particular foraging host (Turlings et al. 1999; Tamò et al. 2006). Olfactometers have been practiced for more than a century. Olfactory measurement is ordinarily used in examinations of parasitoid behavior and responses to olfactory stimuli by some treatments (Pham-Délegue et al. 1991). A parasitoid that could walk into one of the four arms was supposed to have a predilection for the odour introduced through that arm (Sullivan et al. 2000). The four-arm olfactometer was improved by Vet et al. (1983). In this Olfactometer, parasitoids are introduced into a space in which four diverse scent zones are formed. Four-arm olfactometers are well suited for studying the direct behavioural clarifications (Ruther and Steidle 2000). Managing the attraction of these parasitoids is likely to have significant consequences for biocontrol, so jasmonate was applied in this research to investigate whether it affects the attractiveness of Cotesia species. We hypothesize that there are differences in induced volatile emissions between different cotton plant genotypes as well as jasmonate- and host damaged- larvae may influence the parasitoid's attractiveness. Infested cotton plants are known to release amounts of induced volatiles. We had compared the emissions of cotton species and jasmonate treatments in terms of their attractiveness to C. marginiventris and behavior of parasitoids to choose among odors of the three species as well as the control group. 2. MARTIALS & METHODS 2.1. Cotton plants Variety DPL 422 B/R (GM) and 420R (non-GM) (provided by Deltapine company, Mississippi, USA) had been grown in 8cm diameter pots, with a suitable potting soil and positioned in a microclimate chamber (controlled on 25°C, 60% RH, 16:8 h L/D). Plants used for the olfactometer experiments were 3–4 weeks old and had 3–5 fully developed leaves. Four experimental plants were used in four-arm-olfactometer tubes, for each replication, the plants were renewed and replicated 8 times. 2.2. Experiments The experimental approach and stimuli contrasts were carried out to assess different jasmonate as well damaged- larvae on GM and non GM cotton types. To study evaluating the attraction of female C. marginiventris to four different scent source species (Bt with Ja, BT without Ja, Non Bt with Ja and Non Bt without Ja) and to study the parasitoids' preference for a specific odor source; Then the treatments were compared, a-priority contrasts, to the main effects (Jasmonate and cotton type) and make directed comparisons (BtJ vs BtnJ; nBtJa vs nBtnJ) were done. 2.3. Odor sources In this experiment; the treated plants were sprayed with jasmonate (jasmonic acid, purchased from Aldrich Chemical Company, USA). Cotton plants − 4-5-week old were treated with Jasmonate. The spray mixture was prepared by mixing 3 mL of Jasmonate in 8 L of water by adding 3 mL of acetone. Approximately, each plant received about 2.5 micron of jasmonate, according to Thaler (1999). Post three days of application, the plants used in 4-arms olfactometer experiments were to study the attractiveness of female wasps C. marginiventris . Spodoptera exigua used in these experiments were gotten from a laboratory culture sustained in growing chambers (25 ± 2°C, 60–70% RH and a 14: 10 (L: D)) reared according to Perkins (1979). One night before the experiments (ca. 12 h), plants were relocated to glass vessels (Turlings et al. 2004), then infested with one S. exigua larva (2nd instar). For the other treatment, cotton plants were inoculated with 2 larvae before the Olfactometer assay. For control plants; solution contained 3 ml of acetone mixed in 8 L of distilled water; these control plants renewed and replicated 8 times. 2.4. Cotesia marginiventris parasitoids At the Biocontrol Laboratory at Texas A&M University, a primary colony of C. marginiventris was obtained from the USDA, Arizona. As these parasitoids were started to mass reared on the larvae of the beet armyworm (Wiedenmann, 1992), there are adequate wasp numbers to conduct these experiments and to maintain the rearing of the parasitoid in the laboratory for subsequent experiments. Just the beet armyworm larvae have been parasitized by female C. marginiventris ; parasitized larvae were continued in growth chambers as described above. After 10–12 days, parasitoid pupae were collected and kept at 25°C in glass vials until parasitoid adults emerged. The emerging adults were placed in small vials and separated into females and males. Then, females that were 1–2 days old were used in the planned experiments. 2.5. Olfactometer system The four-arm device chambers are designed to be symmetrical, with equal airflow and passage rates through all four channels, it is expected that an even airflow field will prevent air mixing among channels (Vet et al. 1983; Jembere et al. 2003). A four-armed olfactory meter made of 4 Perspex crescents (900 arcs, diameter 27 cm), according to that of Vet et al (1983), was evaluated in these trials. To maintain the air humidity at 80–90% relative humidity, a container filled with distilled water was used (Fig. 1 ). For the temperature, it was fixed at 22–25 degrees Celsius. The air flow is set to 0.02 L/sec in each arm of the four arms. A female wasp of C. marginiventris was introduced individually into the olfactometer area. Every five replicates, the treated cotton plants were replaced. Forty replicates of 40 Cotesia females per run were used as replicates. The performance of the Olfactometer was evaluated qualitatively and this method relies on visual evaluation connected to a computer screen; where parasitoid movement was monitored with a camera connected to PC. It was video directly and the wasp movement was observed by an author. 2.6. Data collection A primary experiment had been conducted to select the day, when the wasps were more responsive after jasmonate application, to measure the attractiveness of the parasite Cotisia after the first, third or seventh day of treatments after spraying jasmonate. The third day was chosen because the C. marginiventris female responded higher and faster at that day. The attractiveness of Cotisia parasitoids to 4 different scent sources was assessed using the following factors: Searching time (the time spent on one arm showing greater preference for a particular arm over others), and selection time (the time spent on a particular arm compared to the time spent on a particular arm over others) and number of visits (the number of visits to the selected area). Four different odor sources were studied and evaluated in cotton and non-BT cotton experiments as the following treatments: jasmonate, one and two caterpillar infestations, and comparison (untreated or non-insect-infested plants). For comparison between GM and non-GM plants treated with and without jasmonate, 4 odorants were examined as the followings: BtJ vs BtnJ; nBtJa vs nBtnJ. 2.7. Data analysis One-way Friedman statistical analysis of variance was used to calculate variance, based on search time, time selection and number of visits for each zone/odor or one of the four arms, to test significance of odor zone preference by female C. marginiventris in olfactometer trials (Zar 1999). Measurement criteria were separated and determined using procedure of Student Newman-Keuls for arranged data (Zar 1999). Data analyzing were done separately for Bt and non-Bt cotton. The analysis of variance of these experiments were done with Tukey's to compare among mean of the treatments, and a-priority contrasts, to the main effects (Jasmonate and cotton type) and make directed comparisons (BtJ vs BtnJ; nBtJa vs nBtnJ) using Statistix 9 (Thomas and Maurice 2008). 3. RESULTS 3.1. Preliminary experiment Cotesia marginiventris females spent 53.3% of the search time in the area where the plant was treated with jasmonate, 68.8% for choosing the time, and 64.3% for the number of visits. While the parasitoid females spent about 26.1% (search), 20.5% (selection) and 23.2% (number of visits) in the arm of plants infested with 2 larvae, while the wasps spent the least time (17.6% for search, 10.7% for selection and 12.5% of visit numbers) in plant areas infested with one larva. In control areas, C. marginiventris females spent only 3.0% searching, while selecting and visits are 0.0% (Fig. 2a). Analyses of data to compare among different treatments; JA, damaged- larvae and control plants showed that there was a significant difference ( P = 0.004). C. marginiventris females took time longer for choosing the suitable arm () on the third day than they did on the first and seventh days. The wasps spent 63.5% of this time in jasmonate areas on the third day, compared to 13.4 and 23.1% on the first and seventh days, respectively. C. marginiventris females spent 47.0% of the time in the area of infested plant by a single larva on the third day, compared to 18.0 and 35.0% on the first and seventh days, respectively; Whereas it spent 51.1% of this time on the third day, compared to 20.8 and 28.1% on the first and seventh days, respectively, in the arms of the plants infested with two larvae zones. There was a significant difference ( P < 0.002 ) in different application dates post JA treatments (Fig. 2b). Cotesia attractiveness on the 3rd day post application responded higher proportion and faster. The volatile profiles analyzed indicated that plants infested with two S. exigua larvae per plant emitted a half or third amount of volatiles as seedlings treated with JA. Therefore, the overall induction resulting from the 2 herbivores was comparable with jasmonate. 3.2. Reactions of Cotesia females to different odors on GM and non-GM-cotton (pooled data) As indicated from the preliminary experiment before C. marginiventris wasps were capable of distinguishing among volatile blends from plants infested with either one or two S. exigua larvae and volatiles released by plants treated with jasmonate. C. marginiventris revealed a strong preference for odors from JA treatment were more attractive than host-infested control plants. In this experiment, female C. marginiventris spent expressively more time searching areas of jasmonate than areas of larval-infested plant, particularly the arms of the two larvae (Fig. 3). C. marginiventris females spent 20.6 and 6.4% in the search area searching the larval infestation arms and control area, respectively. After numerous attempts by the parasitoid females to search in the different four zones, the females chose their favorite odors, and spent different periods of time according to their preferences for these favorite scents in those zones of olfactometer. While C. marginiventris females spent 76.5% of the time selecting in the jasmonate area compared to 17.1% in the larval damage area. Analyze cumulative data using ANOVA to compare JA. Infested caterpillars and control plants showed a significant difference (P = 0.0031). In contrast, wasps spent 6.4% more time selecting into a single larval damage area; 15.2% in the two larvae damage areas, while it was 0.0% in the control area. The number of visits was 71.3% of the complete numbers in the jasmonate zone, 20.4% in the two larval damage region, 8.3% in the one larval damage zone and 0.0% control (Fig. 3). 3.3. Reaction of C. marginiventris females to B. thurigiensis modification in Jasmonate-treated plants Cotesia marginiventris attractiveness was tested for transgenic cotton plants (BT Cotton), where females spent more time foraging in areas that had been sprayed with jasmonate than in areas not treated with jasmonate (Fig. 4). C. marginiventris females expended more time foraging in the arms of the GM cotton than in the non-GM cotton zones (42.0 and 27.0% of the whole time, respectively). On the other hand, female wasps spent 24.0 and 4.9% searching time in non-jasmonate-treated plant areas in GM plants and non-GM plants, respectively. Females of C. marginiventris spent the longest selection time (72.2%) in GM cotton treated with jasmonate zone (GM/JA) compared to (20.5%) in non-BT cotton with jasmonate (NGM/JA) zone. Even though Cotesia wasps spent time in selection (7.3%) in the GM-non-jasmonate cotton zone (GM/NJA) compared to (0.0%) in the non-GM-non-jasmonate cotton zone (NGM/NJA). Similar trend was also observed in the number of visits; whereas C. marginiventris wasps visited the GM cotton areas more than the non-GM areas and visited the arms that had plants treated with jasmonate more than the areas that did not treat by jasmonate (Fig. 4). The analysis of observed data suggests that jasmonate treatments with or without GM plants enhance parasitoid activity compared to non-JA plants and showing significant differences (P < 0.023) between treatments. 4. DISSCUSSION The results of these experiments indicate that C. marginiventris females have been significantly attracted to odors from jasmonate, followed by plants that were fed to S. exigua larvae on both BT and non-BT cotton varieties. The obtained results are analogous to those found by Ozawa et al. (2000, 2004), who reported that volatiles from jasmonate or larvae infestations may influence the success of female C. marginiventris as an active biocontrol agent in controlling these insect pests. Thus, it may be possible to assist the behavior of the parasitoid and support it with plant cynomones and enhance its effectiveness in controlling this insect pest. It has been shown that by identifying the components of plants that attract parasitoid wasps, it could be promising to use this information, for example, to increase the level of these active compounds throughout the cotton breeding and thus increase the efficiency of the parasitoids as mentioned by other authors (Turlings et al. 1991, 2000). Who confirmed that analysis of those volatiles collected from plants, beet armyworm and beet armyworm plant complexes is necessary to define the comparative significance of these informational chemicals in the process of finding insect host by C. marginiventris wasps. There are plant odors that enable natural enemies (parasitoids) to find their insect hosts, by locating the larvae through these odors that originate from the plants after jasmonate application via induced plant volatiles. The same tendency of results was cited by Thaler (1999, 2002); Xiu et al (2019) who itemized that jasmonate is responsible for the induction of many changes in plant resistance that occurs following insect pest attack as well attracts more natural enemies such larval parasitoids. Cotesia marginiventris wasps spent meaningfully more time in areas with jasmonate-treated cotton plants than in areas without jasmonate, as well as in arms containing transgenic cotton plants compared to non-transgenic cotton areas. Similar trend was noticed in the selection of time and number of visits. The present results are consistent with those obtained by El-Wakeil et al. (2003) who stated that experimental plots treated with jasmonate in cotton fields were more attractive to biological control agents with non-jasmonate plots, as well as in GM cotton plots versus non-GM cotton plots. Allied results were confirmed by Ozawa et al (2004), who described that corn plants treated with jasmonic acid attract more parasitoid wasps ( Cotesia kariyai , and Phytoseiulus persimilis ) than control plants. It has been suggested that odorous volatiles caused by herbivores benefit plants in different ways as stated by Vet and Dicke (1992); Dicke and Van Loon (2000); Hob Allah et al. (2002). As many scientists have reported, it has become clear that these odors are important for attracting natural enemies to the microenvironment of their prey and eliminating them biologically (Dicke et al. 1990a; Turlings et al. 1990; Steinberg et al. 1993; Mumm and Dicke 2010; Williams et al. 2017; Ayelo et al. 2021; Ali et al. 2022). This vigorous role of the plant in significant the success rate of host or prey site has been predictable as one aspect that needs to be included in the calculation of possible changes in genetic modified plants as reported by Turlings et al. (2005). Other studies conducted by Schuler et al (1999a, 2003); they revealed that volatiles, which released from caterpillar-damaged leaves, help for finding the host location by parasitoid wasp Cotesia plutellae , especially on the transgenic plants. In other similar studies by Erb et al. (2010), they reported that plant defense responses to insect infestations can be stronger than assumed, in self-defenses, and that volatiles released from infested plants convey specific information about the type of insect attacking, even in the different conditions are complicated with the presence of many herbivores. Thus, these results supported the idea that volatiles caused by herbivores may be one of the indirect plant protection policies from these insect infestations. Kos et al. (2009); Åhman et al (2011); Huang et al (2022) mentioned that transgenic plants enhance the volatile induction and cause a drastic change in volatile structures as reported by Schnee et al (2006), who stated that after infesting transgenic maize relief volatile mixtures that is exceedingly attractive to parasitic wasps. This study offers an informative sample of the value of genetic modified plants in dividing natural interactions facilitated by volatile indicators as reported by Ye et al (2013). This study proposed an evidence for several strategies that induced plants by either jasmonate or larval damage affecting the efficiency of parasitoid wasps to controlling the serious insect pests; as confirmed by Thaler (2002), who confirmed that induced plants could act as defenses against some insect pests by increasing the efficiency of the parasitoid wasps. 5. CONCLUSIONS It is expected, after this knowledge and experiments, that the induction of jasmonate to volatiles is a very useful and effective tool to increase the attractiveness of parasitoids, particularly C. marginiventris , to their insect hosts to benefit from these effective biocontrol agents in organic farms of cotton to sustain product quality and environmental cleanliness. The genetic modification approach to modifying cotton plants to increase volatiles, which enhances attraction to natural enemies, also allows for fundamental changes in the composition of volatiles that play an effective role in activating biological control agents. Furthermore, to make transgenic resistance programs successful, it will be possible to select inducible promoters that release these substances with specific expression in plant tissues infested or treated by jasmonate. Further understanding of signaling compounds and recognition of their molecular structure by recipient plants will allow the development of specific stimuli; promoters used in gene constructs to modify volatile emissions. While developing new transgenic plants in plant breeding programs, it will be necessary to study the main as well as side effects on other organisms in the food web as confirmed by various studies around the world and to ensure that the changes introduced do not reduce the quality of the crops as food or feed. Abbreviations Ja: Jasmonate/ Jasmonic acid BT: Cotton plants are modified with Bacillus thuringiensis NBT: Cotton plants aren’t modified with Bacillus thuringiensis GM: genetic modified BtJ: modified BT plants treated with Jasmonate nBtJ: not modified BT plants treated with Jasmonate Searching time: Parasitoids walk into one of 4 armes show a significant preference for a precise arm. Selecting time : A parasitoid selects an arm with an odour is more specific than in the other choices. Selecttm: Number of visits to the selected zone Tmt: Treatment Declarations Availability of Data and Material Datasets generated for this study can be obtained from the corresponding author. Competing interests The authors declare that they have no competing interests. Funding This research was funded by Deanship of Scientific Research, King Faisal University, Saudi Arabia. Authors details 1 Arid Land Agriculture Department, College of Agricultural and Food Sciences, King Faisal University, Al-Ehsa, Saudi Arabia 2 Pests and Plant Protection Department, National Research Centre, Dokki, Cairo, Egypt Ethics approval and consent to participate : Not applicable Consent for publication: Not applicable ACKNOWLEDGMENTS The authors would like to thank the Deanship of Scientific Research, King Faisal University, Saudi Arabia for the financial support provided to conduct and publish the disclosed research. We would like to thank Prof. J. Bernal (Texas A&M University) for providing us place and facilities to conduct a part of this research in his institute and for useful comments on this MS. The authors are grateful to Dr. Bokonon-Ganta for his procedural help in Olfactometer system, Dr. Setamou for his assistance in the statistical analysis are appreciated. References Åhman I, Glinwood R, Ninkovic V (2011) The potential for modifying plant volatile composition to enhance resistance to arthropod pests. CAB Rev: Perspect in Agric, Vet Sci, Nutri & Nat Res, 5 (6):1-10. https://doi.org/10.1079/PAVSNNR20105006 Ali J, Covaci AD, Roberts JM, Sobhy IS, Kirk WDJ, Bruce TJA (2021a) Effects of Cis-Jasmone treatment of Brassicas on interactions with Myzus Persicae aphids and their parasitoid Diaeretiella Rapae. Front Plant Sci 12, 711896. https://doi.org/10.3389/fpls.2021.711896 Ali MY, Naseem T, Arshad M, Ashraf I, Rizwan M, Tahir M, Rizwan M, Sayed S, Ullah MI, Khan RR, et al. (2021b) Host-Plant variations affect the biotic potential, survival, and population projection of Myzus Persicae (Hemiptera: Aphididae). Insects 12, 375. https://doi.org/10.3390/insects12050375 Ali MY, Naseem T, Holopainen JK, Liu T, Zhang J, Zhang F (2023) Tritrophic interactions among arthropod natural enemies, herbivores and plants considering volatile blends at different scale levels. Cells 12, 251. https://doi.org/10.3390/cells12020251 Ali MY, Naseem,T, Zhang,J, Pan M, Zhang F, Liu T (2022) Plant volatiles and herbivore induced plant volatiles from Chili Pepper act as attractant of the aphid parasitoid. Plants, 11, 1350. https://doi.org/10.3390/plants11101350 Ayelo PM, Pirk CWW, Yusuf AA, Chailleux A, Mohamed SA, Deletre E (2021) Exploring the kairomone-based foraging behaviour of natural enemies to enhance biological control: A Review. Front Ecol Evol 9, 641974. https://doi.org/10.3389/fevo.2021.641974 Clavijo Mccormick A, Gershenzon J, Unsicker SB (2014) Little peaks with big effects: establishing the role of minor plant volatiles in plant–insect interactions. Plant, Cell & Enviro (Special Issue: Plant Volatiles) 37:1836-1844. https://doi.org/10.1111/pce.12357 Clavijo McCormick A, Unsicker SB, Gershenzon J (2012) The specificity of herbivore-induced plant volatiles in attracting herbivore enemies. Trends in Plant Sci17:303–310. https://doi.org/10.1016/j.tplants.2012.03.012 D'Alessandro M, Turlings TCJ (2005) In Situ modification of herbivore-induced plant odours: A novel approach to study the attractiveness of volatile organic compounds to parasitic wasps. Chem Sen30:739-753. https://doi.org/10.1093/chemse/bji066 D'Alessandro M, Turlings TCJ (2006) Advances and challenges in the identification of volatiles that mediate interactions among plants and arthropods. The Analyst131:24-32. https://pubs.rsc.org/en/content/articlehtml/2006/an/b507589k de Boer JG, Hordijk CA, Posthumus MA, Dicke M (2008) Prey and non-prey arthropods sharing a host plant: Effects on induced volatile emission and predator attraction. J Chem Ecol34:281–290. https://link.springer.com/article/10.1007/s10886-007-9405-z Dicke M, Van Loon JJA (2000) Multitrophic effects of herbivore-induced plant volatiles in an evolutionary context. Ento Exp App 97:237–249. https://doi.org/10.1046/j.1570-7458.2000.00736.x Dicke M, Gols R, Ludeking D, Posthumus MA (1999) Jasmonic acid and herbivory differentially induce carnivore attracting plant volatiles in lima bean plants. J Chem Ecol 25:1907–1922. https://link.springer.com/article/10.1023/A:1020942102181 Dicke M, Sabelis MW, Takabayashi J, Bruin J, Posthumus MA (1990a) Plant strategies of manipulating predator-prey interactions through allelochemicals: prospects for application in pest control. J Chem Ecol 16:3091-3118. https://link.springer.com/article/10.1007/BF00979614 Dicke M, van Beek TA, Posthumus MA, Ben Dom N, van Bokhoven H, de Groot AE (1990b) Isolation and identification of volatile kairomone that affects acarine predator-prey interactions - involvement of host plant in its production. J Chem Ecol 16:381-396. DOI: 10.1007/BF01021772 Elbehery H, Saleh MME, El-Wakeil NE (2020) Propagation and application of larval parasitoids. Published in book entitled Cottage Industry of Biocontrol Agents and Their Applications (Elwakeil et al. (eds.)) ISBN 978-3-030-33160-3, © Springer Nature Switzerland AG 2020, 49-71 pp. https://doi.org/10.1007/978-3-030-33161-0_2 El-Wakeil NE (2003) New aspects of biological control of Helicoverpa armigera in organic cotton production. Ph.D. dissertation, Göttingen University, Germany, 140 pp https://ediss.uni-goettingen.de/handle/11858/00-1735-0000-0006-AB51-6 El-Wakeil NE, Bernal J, Vidal S (2003) Effects of jasmonate applications on pest and natural enemy recruitment in cotton fields. Proc the World Cotton Research Conference-3, from 9 to 13 March 2003. Cape Town, South Africa pp; 1239-1248. Erb M, Foresti N, Turlings TCJ (2010) A tritrophic signal that attracts parasitoids to host-damaged plants withstands disruption by non-host herbivores. BMC Plant Biology 10:247- doi:10.1186/1471-2229-10-247 Gu, H, Dorn S (2000) Genetic variation in behavioural response to herbivore-infested plants in the parasitic wasp, Cotesia glomerata (L.). J Insect Behav 13:141-156. https://link.springer.com/article/10.1023/A:1007723811573 Halitschke R, Kessler A, Kahl J, Lorenz A, Baldwin IT (2000) Ecophysiological comparison of direct and indirect defenses in Nicotiana attenuata . Oecologia 124: 408-417. Hoballah MEF, Tamo C, Turlings TCJ (2002) Differential attractiveness of induced odors emitted by eight maize varieties for the parasitoid Cotesia marginiventris : is quality or quantity important? J Chem Ecol 28:951-968. https://link.springer.com/article/10.1023/A:1015253600083 Huang X, Kou J, Jing W, Han X, Liu D, Ghasemzadeh S, Sun P, Shi W, Zhang Y (2022) Transcriptomic and metabolomic reprogramming in cotton after Apolygus lucorum feeding implicated in enhancing recruitment of the parasitoid Peristenus spretus . J Pest Sci, 95:249–262. https://doi.org/10.1007/s10340-021-01369-0 Jembere B, Ngi-Song AJ , Overholt W (2003) Olfactory responses of Cotesia flavipes (Hymenoptera: Braconidae) to target and non-target Lepidoptera and their host plants. Biol Cont 28:360-367. https://doi.org/10.1016/S1049-9644(03)00092-6 Kessler A, Baldwin IT (2001) Defensive function of herbivore-induced plant volatile emissions in nature. Science 291:2141-2144. https://www.science.org/doi/abs/10.1126/science.291.5511.2141 Kos M, van Loon JJA, Dicke M, Vet LEM (2009) Transgenic plants as vital components of integrated pest management. Trends in Biotech 27:621-27. https://doi.org/10.1016/j.tibtech.2009.08.002 Liu J, Zhu J, Zhang P, Han L, Reynolds OL, Zeng R, Wu J, Shao Y, You M, Gurr GM (2017) Silicon supplementation alters the composition of herbivore induced plant volatiles and enhances attraction of parasitoids to infested rice plants. Frontiers Plant Sci 8:1265. https://doi.org/10.3389/fpls.2017.01265 Loughrin JH, Manukian A, Heath RR, Tumlinson JH (1995) Volatiles emitted by different cotton varieties damaged by feeding beet armyworm larvae. J Chem Ecol 21:1217-1227. https://link.springer.com/article/10.1007/BF02228321 Mumm R, Dicke M (2010) Variation in natural plant products and the attraction of bodyguards involved in indirect plant defense. Can J Zool 88:628–667. https://doi.org/10.1139/Z10-032 Ngumbi EN, Ngi-Song AJ, Njagi ENM, Torto R, Wadhams LJ, Birkett MA, Pickett JA, Overholt WA, Torto B (2005) Responses of the stem borer larval endoparasitoid Cotesia flavipes to plant derived synomones: Laboratory and field cage experiments. Biocont Sci, Technol 15:271-279. https://doi.org/10.1080/09583150400016985 Ozawa R, Arimura G, Takabayashi J, Shimoda T, Nishioka T (2000) Involvement of jasmonate- and salicylate-related signaling pathways for the production of specific herbivore-induced volatiles in plants. Plant, Cell Physiol 41:391-398. https://doi.org/10.1093/pcp/41.4.391 Ozawa R, Shiojiri K, Sabelis MW (2004) Corn plants treated with jasmonic acid attract more specialist parasitoids, Thereby increasing parasitization of the common armyworm. J Chem Ecol 30:1797-1808. https://link.springer.com/article/10.1023/B:JOEC.0000042402.04012.c7 Pålsson J, Porcel M, Dekker T, Tasin M (2022) Attract, reward and disrupt: Responses of pests and natural enemies to combinations of habitat manipulation and semiochemicals in organic apple. J Pest Sci 95:619–631. https://link.springer.com/article/10.1007/s10340-021-01379-y Perkins WD (1979) Laboratory rearing of the fall armyworm. The Florida Entomol 62: 81-92. https://www.jstor.org/stable/3494084 Pham-Délegue MH, Trouiller J, Bakchine B, Roger B, Masson C (1991) Age dependency of worker bee response to queen pheromone in a four-armed olfactometer. Insects Sociaux38:283–292. https://link.springer.com/article/10.1007/BF01314914 Pickett JA, Glinwood RT (2007) Chemical ecology. In: van Emden HF, Harrington R, editors. Aphids as crop pests. Trowbridge, UK: CAB Internat pp 235-260. Piesik D. Bocianowski J. Kotwica K. Lema G. Piesik M. Ruzsanyi V. Mayhew CA (2022) Responses of adult Hypera rumicis L. to synthetic plant volatile blends. Molecules 27, 6290. https://doi.org/10.3390/molecules27196290 Pinto-Zevallos D M, Strapasson P, Zarbin PHG (2016) Herbivore induced volatile organic compounds emitted by maize: electrophysiological responses in Spodoptera frugiperda females. Phytochem Lett 16:70–74. doi: 10.1016/j.phytol.2016.03.005 Price PW, Bouton CE, Gross P, McPheron BA, Thompson JN, Weis AE (1980) Interactions among three trophic levels: Influence of plants on interactions between insect herbivores and natural enemies. Ann Rev Ecol, System 11:41–65. https://doi.org/10.1146/annurev.es.11.110180.000353 Ruther J, Steidle JLM (2000) Mites as matchmakers: semiochemicals from host-associated mites attract both sexes of the parasitoid Lariophagus distinguendus. J Chem Ecol 26:1205–1217. https://link.springer.com/article/10.1023/A:1005484027559 Schnee C, Tobias Köllner G, Held M, Turlings TCJ, Gershenzon J, Degenhardt J (2006) The products of a single maize sesquiterpene synthase form a volatile defense signal that attracts natural enemies of maize herbivores. PNAS 103:1129–1134. https://doi.org/10.1073/pnas.0508027103 Schuler TH, Poppy GM, Kerry BR, Deholm I (1999a) Potential side effects of insect -resistant transgenic plants on arthropod natural enemies. Trends in Biotechnol 17:210-216. https://doi.org/10.1016/S0167-7799(98)01298-0 Schuler TH, Potting RPJ, Denholm I, Poppy GM (1999b) Parasitoid behaviour and Bt plants. Nature 400: 825–826. https://www.nature.com/articles/23605 Schuler TH, Potting RPJ, Denholm I, Clark SJ, Clark AJ, Stewart CN et al. (2003) Tritrophic choice experiments with Bt plants, the diamondback moth ( Plutella xylostella ) and the parasitoid Cotesia plutellae . Trans Res 12:351–361. https://link.springer.com/article/10.1023/A:1023342027192 Schuman MC, Barthel K, Baldwin IT (2012) Herbivory-induced volatiles function as defenses increasing fitness of the native plant Nicotiana attenuata in nature. Elife 1:e00007. https://elifesciences.org/articles/00007 Selig P, Keough S, Nalam VJ, Nachappa P (2016) Jasmonate-dependent plant defenses mediate soybean thrips and soybean aphid performance on soybean. Arthropod-Plant Interact 10:273–282. https://link.springer.com/article/10.1007/s11829-016-9437-9 Snoeren TA L, Mumm R, Poelman EH , Yang Y, Pichersky E, Dicke M (2010) The Herbivore-Induced plant volatile methyl salicylate negatively affects attraction of the Parasitoid Diadegma semiclausum . J Chem Ecol 36:479–489. https://link.springer.com/article/10.1007/s10886-010-9787-1 Steinberg S, Dicke M, Vet LEM (1993) Relative importance of infochemicals from first and second trophic level in long-range host location by the larval parasitoid Cotesia glomerata . J Chem Ecol 19:47–59. https://link.springer.com/article/10.1007/BF00987470 Sullivan BT, Pettersson EM, Seltmann KC, Berisford CW (2000) Attraction of the bark beetle parasitoid Roptrocerus xylophagorum (Hymenoptera: Pteromalidae) to host associated olfactory cues. Environ Entomol 29:1138–1151.https://doi.org/10.1603/0046-225X-29.6.1138 Takemoto H, Takabayashi J (2015) Parasitic wasps Aphidius ervi are more attracted to a blend of host-induced plant volatiles than to the independent compounds. J Chem Ecol 41: 801–807. https://link.springer.com/article/10.1007/s10886-015-0615-5 Tamò C, Ricard I, Held M, Davison AC, Turlings TCJ (2006) A comparison of naïve and conditioned responses of three generalist endoparasitoids of lepidopteran larvae to host-induced plant odours. Animal Biol 56:205-220. https://brill.com/view/journals/ab/56/2/article-p205_7.xml Thaler JS (1999) Induced resistance in agricultural crops: Effects of jasmonic acid on herbivory and yield in tomato plants. Environ Entomol 28:30-37. https://doi.org/10.1093/ee/28.1.30 Thaler JS (2002) Effect of jasmonate-induced plant responses on the natural enemies of herbivores. J Anim Ecol 71:141–150. https://doi.org/10.1046/j.0021-8790.2001.00586.x Thomas C, Maurice SC (2008) Statistix 9, Ninth Edition, Managerial Economics McGraw- Hill /Irwin. (ISBN: 0073402818). Turlings TCJ, Erb M (2018) Tritrophic interactions mediated by herbivore-induced plant volatiles: Mechanisms, ecological relevance, and application potential. Ann Rev Entomol 63:433–452. https://doi.org/10.1146/annurev-ento-020117-043507 Turlings TCJ, Scheepmaker JWA, Vet LEM, Tumlinson JH, Lewis WJ (1990) How contact foraging experiences affect the preferences for host –related odors in the larval parasitoid Cotesia marginiventris . J Chem Ecol 16:1577-1589. https://link.springer.com/article/10.1007/BF01014091 Turlings TCJ, Tumlinson JH, Eller FJ, Lewis WJ (1991) Larval-damaged plants: source of volatile synomes that guide the parasitoid Cotesia marginiventris to the microhabitat of its host. Ent Exp App 58:75-82. https://doi.org/10.1111/j.1570-7458.1991.tb01454.x Turlings TCJ, Alborn HT, Loughrin JH, Tumlinson JH (2000) Volicitin, an elicitor of maize volatiles in oral secretion of Spodoptera exigua : isolation and bioactivity. J Chem Ecol 26:189-201. https://link.springer.com/article/10.1023/A:1005449730052 Turlings TCJ, Davison AC, Tamo C (2004) A six-arm olfactometer permitting simultaneous observation of insect attraction and odour trapping. Physiol Entomol 29:45-55. https://doi.org/10.1111/j.1365-3032.2004.0362.x Turlings TCJ, Jeanbourquin PM, Held M, Degen T (2005) Evaluating the induced-odour emission of a Bt maize and its attractiveness to parasitic wasps. Transg Res 14:807–816. https://link.springer.com/article/10.1007/s11248-005-0008-6 Vet LEM, Dicke M (1992) The ecology of infochemical use by natural enemies of herbivores in a tritrophic context. Ann Rev Entomol 37:141–172. Vet LEM, Van Lenteren JC, Heymans M, Meelis E (1983) An airflow olfactometer for measuring olfactory responses of hymenopterous parasitoids and other small insects. Physiol Entomol 8:97–106. https://doi.org/10.1111/j.1365-3032.1983.tb00338.x Wei J, Wang L, Zhao J, Li C, Ge F, Kang L (2011) Ecological tradeoffs between jasmonic acid-dependent direct and indirect plant defences in tritrophic interactions. New Phytolist Trust 189:557–567. https://nph.onlinelibrary.wiley.com/doi/10.1111/j.1469-8137.2010.03491.x Wiedenmann RN, Jr. Smith JW, Darnell PW (1992) Laboratory rearing and biology of the parasite Cotesia flavipes using Diatraea saccharalis as a host. Environ Entomol 21:1160-1167. https://doi.org/10.1093/ee/21.5.1160 Williams L, Rodriguez-Saona C, Castle del Conte SC (2017) Methyl jasmonate induction of cotton: a field test of the ‘attract and reward’ strategy of conservation biological control. AoB PLANTS, 9 (5), plx032. https://doi.org/10.1093/aobpla/plx032 Xiu C, Dai W, Pan H et al (2019) Herbivore-induced plant volatiles enhance field-level parasitism of the mirid bug Apolygus lucorum . Biol Control 135:41–47. https://doi.org/10.1016/j.biocontrol.2019.05.004 Ye M, Song Y, Long J, Wang R, Baerson SR, Pan Z, et al. (2013) Priming of jasmonate-mediated antiherbivore defense responses in rice by silicon. PNAS- USA, 110:E3631–E3639. doi: 10.1073/pnas.1305848110 . Ye M, Veyrat N, Xu H, Hu L, Turlings TCJ, Erb M (2018) An herbivore-induced plant volatile reduces parasitoid attraction by changing the smell of caterpillars. Sci. Adv. 4, eaar4767. https://www.science.org/doi/full/10.1126/sciadv.aar4767 Zar JH: Biostatistical analysis. Fourth edition, Prentice Hall, Upper Saddle River, New Jersey, USA, 663 pp (1999). 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-5306734","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":377515273,"identity":"66bc1724-84b7-45c0-830d-0cd92bff52d4","order_by":0,"name":"NABIL EL-WAKEIL","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA60lEQVRIiWNgGAWjYFAC5gYGhgMMPHzsbQwfgFzGBsJaGCFa2HiOMc4gSQsDm0QakVrM2RvbJH6csZFhk3yW2MzDYCO74QD7ww/4tFj2HGyT7LmRxsMmnXYQqCXNeMMBHmMJfFoMbiS23eD5cBioJb39MQ/D4USgFgb8Wu4/bLv558N/HjbJ441AW/4DtbA//oHfFsa22zw3DvCwSbCBHHYAqIXBDL8tZxLbf8ucSQYGclpi4xyDZOOZh3nMLPBqOX74sOGbY3b2/OzHDBveVNjJ9h1vf3wDnxZ0E4CYmQT1o2AUjIJRMAqwAwBOW04ZGW6xgQAAAABJRU5ErkJggg==","orcid":"","institution":"King Faisal University","correspondingAuthor":true,"prefix":"","firstName":"NABIL","middleName":"","lastName":"EL-WAKEIL","suffix":""},{"id":377515274,"identity":"1fcb3916-be4a-41e7-815e-ad8a844c078f","order_by":1,"name":"NAWAL GAAFAR","email":"","orcid":"","institution":"National Research Centre","correspondingAuthor":false,"prefix":"","firstName":"NAWAL","middleName":"","lastName":"GAAFAR","suffix":""},{"id":377515275,"identity":"1dfb6c84-4a1b-4fd5-beef-f508b54fa3cb","order_by":2,"name":"SAAD ALKAHTANI","email":"","orcid":"","institution":"King Faisal University","correspondingAuthor":false,"prefix":"","firstName":"SAAD","middleName":"","lastName":"ALKAHTANI","suffix":""}],"badges":[],"createdAt":"2024-10-21 19:38:12","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-5306734/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-5306734/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":69023638,"identity":"cfdfab10-5e1e-4c71-be11-448891460db4","added_by":"auto","created_at":"2024-11-14 16:36:15","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":198748,"visible":true,"origin":"","legend":"\u003cp\u003eOlfactometer System\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-5306734/v1/2cb71bb6fefe9ad661afc28f.png"},{"id":69023642,"identity":"cac366ba-3e02-43cd-aa65-ebbbd6c9f7f8","added_by":"auto","created_at":"2024-11-14 16:36:15","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":213788,"visible":true,"origin":"","legend":"\u003cp\u003eReactions of \u003cem\u003eCotesia marginiventris \u003c/em\u003efemales to 4 odors in a primary experiment, (A) Iasmonate or larvae-induced and control (B) Effect of days after application. Different letters indicate significant differences.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-5306734/v1/89bd701f8e5c2a04c8d7c4ec.png"},{"id":69023640,"identity":"5eca257a-b359-45f4-b123-21d6e42177d6","added_by":"auto","created_at":"2024-11-14 16:36:15","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":115402,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cem\u003eCotesia marginiventris \u003c/em\u003efemale’s reactions to four odors on pooled data of BT and non-BT-cotton plants, jasmonate- treated or larvae-damaged; different letters indicate significant differences.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-5306734/v1/7e660f614090fd117bbfe197.png"},{"id":69024504,"identity":"37494186-f866-48c5-a17a-b7a0bd211023","added_by":"auto","created_at":"2024-11-14 16:44:15","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":75375,"visible":true,"origin":"","legend":"\u003cp\u003eReactions of\u003cem\u003eCotesia marginiventris \u003c/em\u003efemale to four odors on GM compared to non-GM-cotton plants treated or non-treated with Jasmonate; different letters indicate significant differences.\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-5306734/v1/bac4befd2ef8d961fe4481b3.png"},{"id":75289881,"identity":"774a704f-c825-47c8-a607-8a9064724d74","added_by":"auto","created_at":"2025-02-03 05:26:55","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1288519,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-5306734/v1/9ef22925-56a0-4d4a-ac4f-235f7ed688f6.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Effectiveness of jasmonate stimulation on attraction of Cotesia marginiventris via olfactory cues on BT- and non-BT-cotton plants","fulltext":[{"header":"1. INTRODUCTION","content":"\u003cp\u003eThe transgenic cotton plants produce a type of BT toxin that helps providing high levels of resistance to insect groups such as Lepidoptera and Coleoptera that feed on important economic crops, and may have side effect on natural enemies (Schuler 1999 a,b; El-Wakeil et al. 2003). The expected effect of these genetic modified plant groups handled with jasmonate on biocontrol agents in general and on parasitoid wasps in particular, has not yet acknowledged the sufficient attention (Gu and Dorn 2000; Turlings et al. 2005). There is a gab to understand effectiveness of Bt cotton with effect of Jasmonic acid and their induced volatiles for attracting the biological control agents.\u003c/p\u003e \u003cp\u003eIn the case of plant volatiles, little is known about their beneficial biological roles for plants. While it is clear that their function is not limited to attracting parasitoids or predators (Mumm and Dicke 2010; Clavijo McCormick et al. 2012); there are key components that are common across plant species. Single compounds may attract natural enemies to foliage damaged by herbivores. (Bruce et al. 2005; Clavijo McCormick et al. 2012; Ali et al. 2022, 2023). In another case, the mixture of small compounds might tell several parasitoids where their host insect pests are located and help killing them biologically (Clavijo McCormick et al. 2014; Ali et al. 2021a).\u003c/p\u003e \u003cp\u003eIt is scientifically proven that plants produce jasmonic acid after exposure to insect infestations, which leads to increased production of some compounds that participate in resistance against insect pests (de Boer et al. 2008). Jasmonic acid has also been known to be involved in the production of plants to release volatile compounds; insect pests and applying of jasmonic acid to leaves trigger volatile terpenoids that have been specifically linked to attracting natural enemies (Dicke et al. 1999; Mumm and Dicke 2010; Liu et al. 2017). Jasmonic acid applications on plants produce proteinase inhibitors and polyphenol oxidases, which helps reducing the number of insect pests and increasing number of biological control agents in the field. (Thaler 1999; D'Alessandro and Turlings 2005, 2006; Pinto-Zevallos et al. 2016 Ali et al. 2021b).\u003c/p\u003e \u003cp\u003eInsect feeding on the plant helps to modify the levels of volatile substances by producing chemical combinations associated with damage to plant tissues, which increases the induced plant defenses (Selig et al. 2016). These volatile substances increase the attraction of the natural enemies of these insect pests (Kessler and Baldwin 2001; Pickett and Glinwood 2007; Pinto-Zevallos et al. 2016; Turlings and Erb 2018; Xiu et al. 2019). The volatiles apparently released constitutively by plants might attract many natural enemy species (Loughrin et al. 1995; Halitschke et al. 2000; Snoeren et al. 2010; Takemoto and Takabayashi 2015; Piesik et al. 2022). The plant's release of chemical compounds that help stimulate the attraction of natural enemies to insects, therefore, this mechanism is considered an indirect means of defense against plants potentially infested by insects and may lead to insect resistance (Price et al. 1980; Mumm and Dicke 2010; Liu et al. 2017), It is known that insect repellent chemical compounds are a direct mechanism for plant defense, or in other words plant resistance to antitoxins against other insect pests. (Dicke et al. 1990 a,b; Wei et al. 2011; Schuman et al. 2012).\u003c/p\u003e \u003cp\u003e \u003cem\u003eCotesia marginiventris\u003c/em\u003e (Cresson) it is an endo larval parasitoid that parasitizes many Lepidopteran insect pests. It is certain that this parasitoid responds to contact chiromones present in the host's by-products like silk and saliva. It also responds strongly to the host's feces, as well as the damage to the plant host's feeding caused by insect larvae (Turlings et al. 1990, 1991; Ngumbi et al. 2005; P\u0026aring;lsson et al. 2021). A volatile or called volicitin; it is a key source of natural enemy's attraction to infested plants (17-hydroxlinolenoyl-l-Gln) (Liu et al. 2017). This volatile substance (volicitin) was separated, identified, and attempted to be created from the oral secretions of the beet armyworm \u003cem\u003eSpodoptera exigua\u003c/em\u003e (Turlings et al. 2000; El-Wakeil 2003). \u003cem\u003eC. marginiventris\u003c/em\u003e, it is hypothesized that this parasitoid responds normally to volatile substances found in some plants, particularly if fed by \u003cem\u003eS. exigua\u003c/em\u003e larvae. During some foraging experiments, females of these parasitoids seem to learn how to respond to a combination of volatiles and odors specific to a plant compound of a particular foraging host (Turlings et al. 1999; Tam\u0026ograve; et al. 2006).\u003c/p\u003e \u003cp\u003eOlfactometers have been practiced for more than a century. Olfactory measurement is ordinarily used in examinations of parasitoid behavior and responses to olfactory stimuli by some treatments (Pham-D\u0026eacute;legue et al. 1991). A parasitoid that could walk into one of the four arms was supposed to have a predilection for the odour introduced through that arm (Sullivan et al. 2000). The four-arm olfactometer was improved by Vet et al. (1983). In this Olfactometer, parasitoids are introduced into a space in which four diverse scent zones are formed. Four-arm olfactometers are well suited for studying the direct behavioural clarifications (Ruther and Steidle 2000).\u003c/p\u003e \u003cp\u003eManaging the attraction of these parasitoids is likely to have significant consequences for biocontrol, so jasmonate was applied in this research to investigate whether it affects the attractiveness of Cotesia species. We hypothesize that there are differences in induced volatile emissions between different cotton plant genotypes as well as jasmonate- and host damaged- larvae may influence the parasitoid's attractiveness. Infested cotton plants are known to release amounts of induced volatiles. We had compared the emissions of cotton species and jasmonate treatments in terms of their attractiveness to \u003cem\u003eC. marginiventris\u003c/em\u003e and behavior of parasitoids to choose among odors of the three species as well as the control group.\u003c/p\u003e"},{"header":"2. MARTIALS \u0026 METHODS","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1. Cotton plants\u003c/h2\u003e \u003cp\u003eVariety DPL 422 B/R (GM) and 420R (non-GM) (provided by Deltapine company, Mississippi, USA) had been grown in 8cm diameter pots, with a suitable potting soil and positioned in a microclimate chamber (controlled on 25\u0026deg;C, 60% RH, 16:8 h L/D). Plants used for the olfactometer experiments were 3\u0026ndash;4 weeks old and had 3\u0026ndash;5 fully developed leaves. Four experimental plants were used in four-arm-olfactometer tubes, for each replication, the plants were renewed and replicated 8 times.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2. Experiments\u003c/h2\u003e \u003cp\u003eThe experimental approach and stimuli contrasts were carried out to assess different jasmonate as well damaged- larvae on GM and non GM cotton types. To study evaluating the attraction of female \u003cem\u003eC. marginiventris\u003c/em\u003e to four different scent source species (Bt with Ja, BT without Ja, Non Bt with Ja and Non Bt without Ja) and to study the parasitoids' preference for a specific odor source; Then the treatments were compared, a-priority contrasts, to the main effects (Jasmonate and cotton type) and make directed comparisons (BtJ vs BtnJ; nBtJa vs nBtnJ) were done.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e2.3. Odor sources\u003c/h2\u003e \u003cp\u003eIn this experiment; the treated plants were sprayed with jasmonate (jasmonic acid, purchased from Aldrich Chemical Company, USA). Cotton plants \u0026minus;\u0026thinsp;4-5-week old were treated with Jasmonate. The spray mixture was prepared by mixing 3 mL of Jasmonate in 8 L of water by adding 3 mL of acetone. Approximately, each plant received about 2.5 micron of jasmonate, according to Thaler (1999). Post three days of application, the plants used in 4-arms olfactometer experiments were to study the attractiveness of female wasps C. \u003cem\u003emarginiventris\u003c/em\u003e.\u003c/p\u003e \u003cp\u003e \u003cem\u003eSpodoptera exigua\u003c/em\u003e used in these experiments were gotten from a laboratory culture sustained in growing chambers (25\u0026thinsp;\u0026plusmn;\u0026thinsp;2\u0026deg;C, 60\u0026ndash;70% RH and a 14: 10 (L: D)) reared according to Perkins (1979). One night before the experiments (ca. 12 h), plants were relocated to glass vessels (Turlings et al. 2004), then infested with one \u003cem\u003eS. exigua\u003c/em\u003e larva (2nd instar). For the other treatment, cotton plants were inoculated with 2 larvae before the Olfactometer assay. For control plants; solution contained 3 ml of acetone mixed in 8 L of distilled water; these control plants renewed and replicated 8 times.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003e2.4. \u003cem\u003eCotesia marginiventris\u003c/em\u003e parasitoids\u003c/h2\u003e \u003cp\u003eAt the Biocontrol Laboratory at Texas A\u0026amp;M University, a primary colony of C. \u003cem\u003emarginiventris\u003c/em\u003e was obtained from the USDA, Arizona. As these parasitoids were started to mass reared on the larvae of the beet armyworm (Wiedenmann, 1992), there are adequate wasp numbers to conduct these experiments and to maintain the rearing of the parasitoid in the laboratory for subsequent experiments. Just the beet armyworm larvae have been parasitized by female \u003cem\u003eC. marginiventris\u003c/em\u003e; parasitized larvae were continued in growth chambers as described above. After 10\u0026ndash;12 days, parasitoid pupae were collected and kept at 25\u0026deg;C in glass vials until parasitoid adults emerged. The emerging adults were placed in small vials and separated into females and males. Then, females that were 1\u0026ndash;2 days old were used in the planned experiments.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003e2.5. Olfactometer system\u003c/h2\u003e \u003cp\u003eThe four-arm device chambers are designed to be symmetrical, with equal airflow and passage rates through all four channels, it is expected that an even airflow field will prevent air mixing among channels (Vet et al. 1983; Jembere et al. 2003). A four-armed olfactory meter made of 4 Perspex crescents (900 arcs, diameter 27 cm), according to that of Vet et al (1983), was evaluated in these trials. To maintain the air humidity at 80\u0026ndash;90% relative humidity, a container filled with distilled water was used (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). For the temperature, it was fixed at 22\u0026ndash;25 degrees Celsius. The air flow is set to 0.02 L/sec in each arm of the four arms. A female wasp of \u003cem\u003eC. marginiventris\u003c/em\u003e was introduced individually into the olfactometer area. Every five replicates, the treated cotton plants were replaced. Forty replicates of 40 \u003cem\u003eCotesia\u003c/em\u003e females per run were used as replicates. The performance of the Olfactometer was evaluated qualitatively and this method relies on visual evaluation connected to a computer screen; where parasitoid movement was monitored with a camera connected to PC. It was video directly and the wasp movement was observed by an author.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003e2.6. Data collection\u003c/h2\u003e \u003cp\u003eA primary experiment had been conducted to select the day, when the wasps were more responsive after jasmonate application, to measure the attractiveness of the parasite \u003cem\u003eCotisia\u003c/em\u003e after the first, third or seventh day of treatments after spraying jasmonate. The third day was chosen because the \u003cem\u003eC. marginiventris\u003c/em\u003e female responded higher and faster at that day. The attractiveness of \u003cem\u003eCotisia\u003c/em\u003e parasitoids to 4 different scent sources was assessed using the following factors: Searching time (the time spent on one arm showing greater preference for a particular arm over others), and selection time (the time spent on a particular arm compared to the time spent on a particular arm over others) and number of visits (the number of visits to the selected area). Four different odor sources were studied and evaluated in cotton and non-BT cotton experiments as the following treatments: jasmonate, one and two caterpillar infestations, and comparison (untreated or non-insect-infested plants). For comparison between GM and non-GM plants treated with and without jasmonate, 4 odorants were examined as the followings: BtJ vs BtnJ; nBtJa vs nBtnJ.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003e2.7. Data analysis\u003c/h2\u003e \u003cp\u003eOne-way Friedman statistical analysis of variance was used to calculate variance, based on search time, time selection and number of visits for each zone/odor or one of the four arms, to test significance of odor zone preference by female \u003cem\u003eC. marginiventris\u003c/em\u003e in olfactometer trials (Zar 1999). Measurement criteria were separated and determined using procedure of Student Newman-Keuls for arranged data (Zar 1999). Data analyzing were done separately for Bt and non-Bt cotton. The analysis of variance of these experiments were done with Tukey's to compare among mean of the treatments, and a-priority contrasts, to the main effects (Jasmonate and cotton type) and make directed comparisons (BtJ vs BtnJ; nBtJa vs nBtnJ) using Statistix 9 (Thomas and Maurice 2008).\u003c/p\u003e \u003c/div\u003e"},{"header":"3. RESULTS","content":"\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e\n \u003ch2\u003e3.1. Preliminary experiment\u003c/h2\u003e\n \u003cp\u003e\u003cem\u003eCotesia marginiventris\u003c/em\u003e females spent 53.3% of the search time in the area where the plant was treated with jasmonate, 68.8% for choosing the time, and 64.3% for the number of visits. While the parasitoid females spent about 26.1% (search), 20.5% (selection) and 23.2% (number of visits) in the arm of plants infested with 2 larvae, while the wasps spent the least time (17.6% for search, 10.7% for selection and 12.5% of visit numbers) in plant areas infested with one larva. In control areas, \u003cem\u003eC. marginiventris\u003c/em\u003e females spent only 3.0% searching, while selecting and visits are 0.0% (Fig. 2a). Analyses of data to compare among different treatments; JA, damaged- larvae and control plants showed that there was a significant difference (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.004).\u003c/p\u003e\n \u003cp\u003e\u003cem\u003eC. marginiventris\u003c/em\u003e females took time longer for choosing the suitable arm () on the third day than they did on the first and seventh days. The wasps spent 63.5% of this time in jasmonate areas on the third day, compared to 13.4 and 23.1% on the first and seventh days, respectively. \u003cem\u003eC. marginiventris\u003c/em\u003e females spent 47.0% of the time in the area of infested plant by a single larva on the third day, compared to 18.0 and 35.0% on the first and seventh days, respectively; Whereas it spent 51.1% of this time on the third day, compared to 20.8 and 28.1% on the first and seventh days, respectively, in the arms of the plants infested with two larvae zones. There was a significant difference (\u003cspan type=\"SmallCaps\" class=\"SmallCaps\" name=\"Emphasis\"\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.002\u003c/span\u003e) in different application dates post JA treatments (Fig.\u0026nbsp;2b).\u003c/p\u003e\n \u003cp\u003e\u003cem\u003eCotesia\u003c/em\u003e attractiveness on the 3rd day post application responded higher proportion and faster. The volatile profiles analyzed indicated that plants infested with two \u003cem\u003eS. exigua\u003c/em\u003e larvae per plant emitted a half or third amount of volatiles as seedlings treated with JA. Therefore, the overall induction resulting from the 2 herbivores was comparable with jasmonate.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e\n \u003ch2\u003e3.2. Reactions of \u003cem\u003eCotesia\u003c/em\u003e females to different odors on GM and non-GM-cotton (pooled data)\u003c/h2\u003e\n \u003cp\u003eAs indicated from the preliminary experiment before \u003cem\u003eC. marginiventris\u003c/em\u003e wasps were capable of distinguishing among volatile blends from plants infested with either one or two \u003cem\u003eS. exigua\u003c/em\u003e larvae and volatiles released by plants treated with jasmonate. \u003cem\u003eC. marginiventris\u003c/em\u003e revealed a strong preference for odors from JA treatment were more attractive than host-infested control plants.\u003c/p\u003e\n \u003cp\u003eIn this experiment, female \u003cem\u003eC. marginiventris\u003c/em\u003e spent expressively more time searching areas of jasmonate than areas of larval-infested plant, particularly the arms of the two larvae (Fig. 3). \u003cem\u003eC. marginiventris\u003c/em\u003e females spent 20.6 and 6.4% in the search area searching the larval infestation arms and control area, respectively. After numerous attempts by the parasitoid females to search in the different four zones, the females chose their favorite odors, and spent different periods of time according to their preferences for these favorite scents in those zones of olfactometer. While \u003cem\u003eC. marginiventris\u003c/em\u003e females spent 76.5% of the time selecting in the jasmonate area compared to 17.1% in the larval damage area. Analyze cumulative data using ANOVA to compare JA. Infested caterpillars and control plants showed a significant difference (P\u0026thinsp;=\u0026thinsp;0.0031). In contrast, wasps spent 6.4% more time selecting into a single larval damage area; 15.2% in the two larvae damage areas, while it was 0.0% in the control area. The number of visits was 71.3% of the complete numbers in the jasmonate zone, 20.4% in the two larval damage region, 8.3% in the one larval damage zone and 0.0% control (Fig. 3).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv id=\"Sec13\" class=\"Section2\"\u003e\n \u003ch2\u003e3.3. Reaction of \u003cem\u003eC. marginiventris\u003c/em\u003e females to \u003cem\u003eB. thurigiensis\u003c/em\u003e modification in Jasmonate-treated plants\u003c/h2\u003e\n \u003cp\u003e\u003cem\u003eCotesia marginiventris\u003c/em\u003e attractiveness was tested for transgenic cotton plants (BT Cotton), where females spent more time foraging in areas that had been sprayed with jasmonate than in areas not treated with jasmonate (Fig. 4). \u003cem\u003eC. marginiventris\u003c/em\u003e females expended more time foraging in the arms of the GM cotton than in the non-GM cotton zones (42.0 and 27.0% of the whole time, respectively). On the other hand, female wasps spent 24.0 and 4.9% searching time in non-jasmonate-treated plant areas in GM plants and non-GM plants, respectively. Females of \u003cem\u003eC. marginiventris\u003c/em\u003e spent the longest selection time (72.2%) in GM cotton treated with jasmonate zone (GM/JA) compared to (20.5%) in non-BT cotton with jasmonate (NGM/JA) zone. Even though \u003cem\u003eCotesia\u003c/em\u003e wasps spent time in selection (7.3%) in the GM-non-jasmonate cotton zone (GM/NJA) compared to (0.0%) in the non-GM-non-jasmonate cotton zone (NGM/NJA). Similar trend was also observed in the number of visits; whereas \u003cem\u003eC. marginiventris\u003c/em\u003e wasps visited the GM cotton areas more than the non-GM areas and visited the arms that had plants treated with jasmonate more than the areas that did not treat by jasmonate (Fig. 4). The analysis of observed data suggests that jasmonate treatments with or without GM plants enhance parasitoid activity compared to non-JA plants and showing significant differences (P\u0026thinsp;\u0026lt;\u0026thinsp;0.023) between treatments.\u003c/p\u003e\n\u003c/div\u003e"},{"header":"4. DISSCUSSION","content":"\u003cp\u003eThe results of these experiments indicate that \u003cem\u003eC. marginiventris\u003c/em\u003e females have been significantly attracted to odors from jasmonate, followed by plants that were fed to \u003cem\u003eS. exigua\u003c/em\u003e larvae on both BT and non-BT cotton varieties. The obtained results are analogous to those found by Ozawa et al. (2000, 2004), who reported that volatiles from jasmonate or larvae infestations may influence the success of female \u003cem\u003eC. marginiventris\u003c/em\u003e as an active biocontrol agent in controlling these insect pests. Thus, it may be possible to assist the behavior of the parasitoid and support it with plant cynomones and enhance its effectiveness in controlling this insect pest. It has been shown that by identifying the components of plants that attract parasitoid wasps, it could be promising to use this information, for example, to increase the level of these active compounds throughout the cotton breeding and thus increase the efficiency of the parasitoids as mentioned by other authors (Turlings et al. 1991, 2000). Who confirmed that analysis of those volatiles collected from plants, beet armyworm and beet armyworm plant complexes is necessary to define the comparative significance of these informational chemicals in the process of finding insect host by \u003cem\u003eC. marginiventris\u003c/em\u003e wasps.\u003c/p\u003e \u003cp\u003eThere are plant odors that enable natural enemies (parasitoids) to find their insect hosts, by locating the larvae through these odors that originate from the plants after jasmonate application via induced plant volatiles. The same tendency of results was cited by Thaler (1999, 2002); Xiu et al (2019) who itemized that jasmonate is responsible for the induction of many changes in plant resistance that occurs following insect pest attack as well attracts more natural enemies such larval parasitoids.\u003c/p\u003e \u003cp\u003e \u003cem\u003eCotesia marginiventris\u003c/em\u003e wasps spent meaningfully more time in areas with jasmonate-treated cotton plants than in areas without jasmonate, as well as in arms containing transgenic cotton plants compared to non-transgenic cotton areas. Similar trend was noticed in the selection of time and number of visits. The present results are consistent with those obtained by El-Wakeil et al. (2003) who stated that experimental plots treated with jasmonate in cotton fields were more attractive to biological control agents with non-jasmonate plots, as well as in GM cotton plots versus non-GM cotton plots. Allied results were confirmed by Ozawa et al (2004), who described that corn plants treated with jasmonic acid attract more parasitoid wasps (\u003cem\u003eCotesia kariyai\u003c/em\u003e, and \u003cem\u003ePhytoseiulus persimilis\u003c/em\u003e) than control plants.\u003c/p\u003e \u003cp\u003eIt has been suggested that odorous volatiles caused by herbivores benefit plants in different ways as stated by Vet and Dicke (1992); Dicke and Van Loon (2000); Hob Allah et al. (2002). As many scientists have reported, it has become clear that these odors are important for attracting natural enemies to the microenvironment of their prey and eliminating them biologically (Dicke et al. 1990a; Turlings et al. 1990; Steinberg et al. 1993; Mumm and Dicke 2010; Williams et al. 2017; Ayelo et al. 2021; Ali et al. 2022). This vigorous role of the plant in significant the success rate of host or prey site has been predictable as one aspect that needs to be included in the calculation of possible changes in genetic modified plants as reported by Turlings et al. (2005). Other studies conducted by Schuler et al (1999a, 2003); they revealed that volatiles, which released from caterpillar-damaged leaves, help for finding the host location by parasitoid wasp \u003cem\u003eCotesia plutellae\u003c/em\u003e, especially on the transgenic plants.\u003c/p\u003e \u003cp\u003eIn other similar studies by Erb et al. (2010), they reported that plant defense responses to insect infestations can be stronger than assumed, in self-defenses, and that volatiles released from infested plants convey specific information about the type of insect attacking, even in the different conditions are complicated with the presence of many herbivores. Thus, these results supported the idea that volatiles caused by herbivores may be one of the indirect plant protection policies from these insect infestations. Kos et al. (2009); \u0026Aring;hman et al (2011); Huang et al (2022) mentioned that transgenic plants enhance the volatile induction and cause a drastic change in volatile structures as reported by Schnee et al (2006), who stated that after infesting transgenic maize relief volatile mixtures that is exceedingly attractive to parasitic wasps. This study offers an informative sample of the value of genetic modified plants in dividing natural interactions facilitated by volatile indicators as reported by Ye et al (2013).\u003c/p\u003e \u003cp\u003eThis study proposed an evidence for several strategies that induced plants by either jasmonate or larval damage affecting the efficiency of parasitoid wasps to controlling the serious insect pests; as confirmed by Thaler (2002), who confirmed that induced plants could act as defenses against some insect pests by increasing the efficiency of the parasitoid wasps.\u003c/p\u003e"},{"header":"5. CONCLUSIONS","content":"\u003cp\u003eIt is expected, after this knowledge and experiments, that the induction of jasmonate to volatiles is a very useful and effective tool to increase the attractiveness of parasitoids, particularly \u003cem\u003eC. marginiventris\u003c/em\u003e, to their insect hosts to benefit from these effective biocontrol agents in organic farms of cotton to sustain product quality and environmental cleanliness. The genetic modification approach to modifying cotton plants to increase volatiles, which enhances attraction to natural enemies, also allows for fundamental changes in the composition of volatiles that play an effective role in activating biological control agents. Furthermore, to make transgenic resistance programs successful, it will be possible to select inducible promoters that release these substances with specific expression in plant tissues infested or treated by jasmonate. Further understanding of signaling compounds and recognition of their molecular structure by recipient plants will allow the development of specific stimuli; promoters used in gene constructs to modify volatile emissions. While developing new transgenic plants in plant breeding programs, it will be necessary to study the main as well as side effects on other organisms in the food web as confirmed by various studies around the world and to ensure that the changes introduced do not reduce the quality of the crops as food or feed.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cp\u003e\u003cstrong\u003eJa:\u003c/strong\u003e Jasmonate/ Jasmonic acid \u003cstrong\u003eBT:\u003c/strong\u003e Cotton plants are modified with \u003cem\u003eBacillus thuringiensis\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eNBT:\u003c/strong\u003e Cotton plants aren’t modified with \u003cem\u003eBacillus thuringiensis \u003c/em\u003e GM: genetic modified\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eBtJ:\u003c/strong\u003e modified BT plants treated with Jasmonate\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003enBtJ:\u003c/strong\u003e not modified BT plants treated with Jasmonate\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSearching time: \u003c/strong\u003eParasitoids walk into one of 4 armes show a significant preference for a precise arm.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSelecting time\u003c/strong\u003e: A parasitoid selects an arm with an odour is more specific than in the other choices.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSelecttm:\u003c/strong\u003e Number of visits to the selected zone \u003cstrong\u003eTmt:\u003c/strong\u003e Treatment\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAvailability of Data and Material\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eDatasets generated for this study can be obtained from the corresponding author.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no competing interests.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research was funded by\u0026nbsp;Deanship of Scientific Research, King Faisal University, Saudi Arabia.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors details\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003csup\u003e1\u0026nbsp;\u003c/sup\u003eArid Land Agriculture Department, College of Agricultural and Food Sciences, King Faisal University, Al-Ehsa, Saudi Arabia\u003c/p\u003e\n\u003cp\u003e\u003csup\u003e2\u0026nbsp;\u003c/sup\u003ePests and Plant Protection Department, National Research Centre, Dokki, Cairo, Egypt\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate\u003c/strong\u003e\u003cstrong\u003e:\u0026nbsp;\u003c/strong\u003eNot applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication:\u0026nbsp;\u003c/strong\u003eNot applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eACKNOWLEDGMENTS\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors would like to thank the Deanship of Scientific Research, King Faisal University, Saudi Arabia for the financial support provided to conduct and publish the disclosed research. We would like to thank Prof. J. Bernal (Texas A\u0026amp;M University) for providing us place and facilities to conduct a part of this research in his institute and for useful comments on this MS. The authors are grateful to Dr. Bokonon-Ganta for his procedural help in Olfactometer system, Dr. Setamou for his assistance in the statistical analysis are appreciated.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003e\u0026Aring;hman I, Glinwood R, Ninkovic V (2011) The potential for modifying plant volatile composition to enhance resistance to arthropod pests. CAB Rev: Perspect in Agric, Vet Sci, Nutri \u0026amp; Nat Res, 5 (6):1-10. https://doi.org/10.1079/PAVSNNR20105006\u003c/li\u003e\n\u003cli\u003eAli J, Covaci AD, Roberts JM, Sobhy IS, Kirk WDJ, Bruce TJA (2021a) Effects of Cis-Jasmone treatment of Brassicas on interactions with \u003cem\u003eMyzus Persicae\u003c/em\u003e aphids and their parasitoid \u003cem\u003eDiaeretiella Rapae. \u003c/em\u003eFront Plant Sci 12, 711896. https://doi.org/10.3389/fpls.2021.711896\u003c/li\u003e\n\u003cli\u003eAli MY, Naseem T, Arshad M, Ashraf I, Rizwan M, Tahir M, Rizwan M, Sayed S, Ullah MI, Khan RR, et al. (2021b) Host-Plant variations affect the biotic potential, survival, and population projection of \u003cem\u003eMyzus Persicae\u003c/em\u003e (Hemiptera: Aphididae). Insects 12, 375. https://doi.org/10.3390/insects12050375\u003c/li\u003e\n\u003cli\u003eAli MY, Naseem T, Holopainen JK, Liu T, Zhang J, Zhang F (2023) Tritrophic interactions among arthropod natural enemies, herbivores and plants considering volatile blends at different scale levels. Cells 12, 251. https://doi.org/10.3390/cells12020251\u003c/li\u003e\n\u003cli\u003eAli MY, Naseem,T, Zhang,J, Pan M, Zhang F, Liu T (2022) Plant volatiles and herbivore induced plant volatiles from Chili Pepper act as attractant of the aphid parasitoid. Plants, 11, 1350. https://doi.org/10.3390/plants11101350\u003c/li\u003e\n\u003cli\u003eAyelo PM, Pirk CWW, Yusuf AA, Chailleux A, Mohamed SA, Deletre E (2021) Exploring the kairomone-based foraging behaviour of natural enemies to enhance biological control: A Review. Front Ecol Evol 9, 641974. https://doi.org/10.3389/fevo.2021.641974\u003c/li\u003e\n\u003cli\u003eClavijo Mccormick A, Gershenzon J, Unsicker SB (2014) Little peaks with big effects: establishing the role of minor plant volatiles in plant\u0026ndash;insect interactions. Plant, Cell \u0026amp; Enviro (Special Issue: Plant Volatiles) 37:1836-1844. https://doi.org/10.1111/pce.12357\u003c/li\u003e\n\u003cli\u003eClavijo McCormick A, Unsicker SB, Gershenzon J (2012) The specificity of herbivore-induced plant volatiles in attracting herbivore enemies. Trends in Plant Sci17:303\u0026ndash;310. https://doi.org/10.1016/j.tplants.2012.03.012\u003c/li\u003e\n\u003cli\u003eD\u0026apos;Alessandro M, Turlings TCJ (2005) In Situ modification of herbivore-induced plant odours: A novel approach to study the attractiveness of volatile organic compounds to parasitic wasps. Chem Sen30:739-753. https://doi.org/10.1093/chemse/bji066\u003c/li\u003e\n\u003cli\u003eD\u0026apos;Alessandro M, Turlings TCJ (2006) Advances and challenges in the identification of volatiles that mediate interactions among plants and arthropods. The Analyst131:24-32. https://pubs.rsc.org/en/content/articlehtml/2006/an/b507589k\u003c/li\u003e\n\u003cli\u003ede Boer JG, Hordijk CA, Posthumus MA, Dicke M (2008) Prey and non-prey arthropods sharing a host plant: Effects on induced volatile emission and predator attraction. J Chem Ecol34:281\u0026ndash;290. https://link.springer.com/article/10.1007/s10886-007-9405-z\u003c/li\u003e\n\u003cli\u003eDicke M, Van Loon JJA (2000) Multitrophic effects of herbivore-induced plant volatiles in an evolutionary context. Ento Exp App 97:237\u0026ndash;249. https://doi.org/10.1046/j.1570-7458.2000.00736.x\u003c/li\u003e\n\u003cli\u003eDicke M, Gols R, Ludeking D, Posthumus MA (1999) Jasmonic acid and herbivory differentially induce carnivore attracting plant volatiles in lima bean plants. J Chem Ecol 25:1907\u0026ndash;1922. https://link.springer.com/article/10.1023/A:1020942102181\u003c/li\u003e\n\u003cli\u003eDicke M, Sabelis MW, Takabayashi J, Bruin J, Posthumus MA (1990a) Plant strategies of manipulating predator-prey interactions through allelochemicals: prospects for application in pest control. J Chem Ecol 16:3091-3118. https://link.springer.com/article/10.1007/BF00979614\u003c/li\u003e\n\u003cli\u003eDicke M, van Beek TA, Posthumus MA, Ben Dom N, van Bokhoven H, de Groot AE (1990b) Isolation and identification of volatile kairomone that affects acarine predator-prey interactions - involvement of host plant in its production. J Chem Ecol 16:381-396. DOI: 10.1007/BF01021772\u003c/li\u003e\n\u003cli\u003eElbehery H, Saleh MME, El-Wakeil NE (2020) Propagation and application of larval parasitoids. Published in book entitled Cottage Industry of Biocontrol Agents and Their Applications (Elwakeil et al. (eds.)) ISBN 978-3-030-33160-3, \u0026copy; Springer Nature Switzerland AG 2020, 49-71 pp. \u003cu\u003ehttps://doi.org/10.1007/978-3-030-33161-0_2\u003c/u\u003e\u003c/li\u003e\n\u003cli\u003eEl-Wakeil NE (2003) New aspects of biological control of \u003cem\u003eHelicoverpa armigera\u003c/em\u003e in organic cotton production. Ph.D. dissertation, G\u0026ouml;ttingen University, Germany, 140 pp https://ediss.uni-goettingen.de/handle/11858/00-1735-0000-0006-AB51-6\u003c/li\u003e\n\u003cli\u003eEl-Wakeil NE, Bernal J, Vidal S (2003) Effects of jasmonate applications on pest and natural enemy recruitment in cotton fields. Proc the World Cotton Research Conference-3, from 9 to 13 March 2003. Cape Town, South Africa pp; 1239-1248.\u003c/li\u003e\n\u003cli\u003eErb M, Foresti N, Turlings TCJ (2010) A tritrophic signal that attracts parasitoids to host-damaged plants withstands disruption by non-host herbivores. BMC Plant Biology 10:247- \u003cu\u003edoi:10.1186/1471-2229-10-247\u003c/u\u003e\u003c/li\u003e\n\u003cli\u003eGu, H, Dorn S (2000) Genetic variation in behavioural response to herbivore-infested plants in the parasitic wasp, \u003cem\u003eCotesia glomerata\u003c/em\u003e (L.). J Insect Behav 13:141-156. https://link.springer.com/article/10.1023/A:1007723811573\u003c/li\u003e\n\u003cli\u003eHalitschke R, Kessler A, Kahl J, Lorenz A, Baldwin IT (2000) Ecophysiological comparison of direct and indirect defenses in \u003cem\u003eNicotiana attenuata\u003c/em\u003e. Oecologia 124: 408-417.\u003c/li\u003e\n\u003cli\u003eHoballah MEF, Tamo C, Turlings TCJ (2002) Differential attractiveness of induced odors emitted by eight maize varieties for the parasitoid \u003cem\u003eCotesia marginiventris\u003c/em\u003e: is quality or quantity important? J Chem Ecol 28:951-968. https://link.springer.com/article/10.1023/A:1015253600083\u003c/li\u003e\n\u003cli\u003eHuang X, Kou J, Jing W, Han X, Liu D, Ghasemzadeh S, Sun P, Shi W, Zhang Y (2022) Transcriptomic and metabolomic reprogramming in cotton after \u003cem\u003eApolygus lucorum\u003c/em\u003e feeding implicated in enhancing recruitment of the parasitoid \u003cem\u003ePeristenus spretus\u003c/em\u003e. J Pest Sci, 95:249\u0026ndash;262. https://doi.org/10.1007/s10340-021-01369-0\u003c/li\u003e\n\u003cli\u003eJembere B, Ngi-Song AJ\u003csup\u003e,\u003c/sup\u003e Overholt W (2003) Olfactory responses of \u003cem\u003eCotesia flavipes\u003c/em\u003e (Hymenoptera: Braconidae) to target and non-target Lepidoptera and their host plants. Biol Cont 28:360-367. https://doi.org/10.1016/S1049-9644(03)00092-6\u003c/li\u003e\n\u003cli\u003eKessler A, Baldwin IT (2001) Defensive function of herbivore-induced plant volatile emissions in nature. Science 291:2141-2144. https://www.science.org/doi/abs/10.1126/science.291.5511.2141\u003c/li\u003e\n\u003cli\u003eKos M, van Loon JJA, Dicke M, Vet LEM (2009) Transgenic plants as vital components of integrated pest management. Trends in Biotech 27:621-27. https://doi.org/10.1016/j.tibtech.2009.08.002\u003c/li\u003e\n\u003cli\u003eLiu J, Zhu J, Zhang P, Han L, Reynolds OL, Zeng R, Wu J, Shao Y, You M, Gurr GM (2017) Silicon supplementation alters the composition of herbivore induced plant volatiles and enhances attraction of parasitoids to infested rice plants. Frontiers Plant Sci 8:1265. https://doi.org/10.3389/fpls.2017.01265\u003c/li\u003e\n\u003cli\u003eLoughrin JH, Manukian A, Heath RR, Tumlinson JH (1995) Volatiles emitted by different cotton varieties damaged by feeding beet armyworm larvae. J Chem Ecol 21:1217-1227. https://link.springer.com/article/10.1007/BF02228321\u003c/li\u003e\n\u003cli\u003eMumm R, Dicke M (2010) Variation in natural plant products and the attraction of bodyguards involved in indirect plant defense. Can J Zool\u003cem\u003e \u003c/em\u003e88:628\u0026ndash;667. https://doi.org/10.1139/Z10-032\u003c/li\u003e\n\u003cli\u003eNgumbi EN, Ngi-Song AJ, Njagi ENM, Torto R, Wadhams LJ, Birkett MA, Pickett JA, Overholt WA, Torto B (2005) Responses of the stem borer larval endoparasitoid \u003cem\u003eCotesia flavipes\u003c/em\u003e to plant derived synomones: Laboratory and field cage experiments. Biocont Sci, Technol 15:271-279. https://doi.org/10.1080/09583150400016985\u003c/li\u003e\n\u003cli\u003eOzawa R, Arimura G, Takabayashi J, Shimoda T, Nishioka T (2000) Involvement of jasmonate- and salicylate-related signaling pathways for the production of specific herbivore-induced volatiles in plants. Plant, Cell Physiol 41:391-398. https://doi.org/10.1093/pcp/41.4.391\u003c/li\u003e\n\u003cli\u003eOzawa R, Shiojiri K, Sabelis MW (2004) Corn plants treated with jasmonic acid attract more specialist parasitoids, Thereby increasing parasitization of the common armyworm. J Chem Ecol 30:1797-1808. https://link.springer.com/article/10.1023/B:JOEC.0000042402.04012.c7\u003c/li\u003e\n\u003cli\u003eP\u0026aring;lsson J, Porcel M, Dekker T, Tasin M (2022) Attract, reward and disrupt: Responses of pests and natural enemies to combinations of habitat manipulation and semiochemicals in organic apple. J Pest Sci 95:619\u0026ndash;631. https://link.springer.com/article/10.1007/s10340-021-01379-y\u003c/li\u003e\n\u003cli\u003ePerkins WD (1979) Laboratory rearing of the fall armyworm. The Florida Entomol 62: 81-92. https://www.jstor.org/stable/3494084\u003c/li\u003e\n\u003cli\u003ePham-D\u0026eacute;legue MH, Trouiller J, Bakchine B, Roger B, Masson C (1991) Age dependency of worker bee response to queen pheromone in a four-armed olfactometer. Insects Sociaux38:283\u0026ndash;292. https://link.springer.com/article/10.1007/BF01314914\u003c/li\u003e\n\u003cli\u003ePickett JA, Glinwood RT (2007) Chemical ecology. In: van Emden HF, Harrington R, editors. Aphids as crop pests. Trowbridge, UK: CAB Internat pp 235-260. \u003c/li\u003e\n\u003cli\u003ePiesik D. Bocianowski J. Kotwica K. Lema G. Piesik M. Ruzsanyi V. Mayhew CA (2022) Responses of adult \u003cem\u003eHypera rumicis\u003c/em\u003e L. to synthetic plant volatile blends. Molecules 27, 6290. https://doi.org/10.3390/molecules27196290\u003c/li\u003e\n\u003cli\u003ePinto-Zevallos D M, Strapasson P, Zarbin PHG (2016) Herbivore induced volatile organic compounds emitted by maize: electrophysiological responses in \u003cem\u003eSpodoptera frugiperda \u003c/em\u003efemales. Phytochem Lett 16:70\u0026ndash;74. \u003cu\u003edoi: 10.1016/j.phytol.2016.03.005\u003c/u\u003e\u003c/li\u003e\n\u003cli\u003ePrice PW, Bouton CE, Gross P, McPheron BA, Thompson JN, Weis AE (1980) Interactions among three trophic levels: Influence of plants on interactions between insect herbivores and natural enemies. Ann Rev Ecol, System 11:41\u0026ndash;65. https://doi.org/10.1146/annurev.es.11.110180.000353\u003c/li\u003e\n\u003cli\u003eRuther J, Steidle JLM (2000) Mites as matchmakers: semiochemicals from host-associated mites attract both sexes of the parasitoid Lariophagus distinguendus. J Chem Ecol 26:1205\u0026ndash;1217. https://link.springer.com/article/10.1023/A:1005484027559\u003c/li\u003e\n\u003cli\u003eSchnee C, Tobias K\u0026ouml;llner G, Held M, Turlings TCJ, Gershenzon J, Degenhardt J (2006) The products of a single maize sesquiterpene synthase form a volatile defense signal that attracts natural enemies of maize herbivores. PNAS 103:1129\u0026ndash;1134. https://doi.org/10.1073/pnas.0508027103\u003c/li\u003e\n\u003cli\u003eSchuler TH, Poppy GM, Kerry BR, Deholm I (1999a) Potential side effects of insect -resistant transgenic plants on arthropod natural enemies. Trends in Biotechnol 17:210-216. https://doi.org/10.1016/S0167-7799(98)01298-0\u003c/li\u003e\n\u003cli\u003eSchuler TH, Potting RPJ, Denholm I, Poppy GM (1999b) Parasitoid behaviour and Bt plants. Nature 400: 825\u0026ndash;826. https://www.nature.com/articles/23605\u003c/li\u003e\n\u003cli\u003eSchuler TH, Potting RPJ, Denholm I, Clark SJ, Clark AJ, Stewart CN et al. (2003) Tritrophic choice experiments with Bt plants, the diamondback moth (\u003cem\u003ePlutella xylostella\u003c/em\u003e) and the parasitoid \u003cem\u003eCotesia plutellae\u003c/em\u003e. Trans Res 12:351\u0026ndash;361. https://link.springer.com/article/10.1023/A:1023342027192\u003c/li\u003e\n\u003cli\u003eSchuman MC, Barthel K, Baldwin IT (2012) Herbivory-induced volatiles\u003cbr\u003efunction as defenses increasing fitness of the native plant \u003cem\u003eNicotiana attenuata \u003c/em\u003ein nature. \u003cem\u003eElife \u003c/em\u003e1:e00007. https://elifesciences.org/articles/00007\u003c/li\u003e\n\u003cli\u003eSelig P, Keough S, Nalam VJ, Nachappa P (2016) Jasmonate-dependent plant defenses mediate soybean thrips and soybean aphid performance on soybean. Arthropod-Plant Interact 10:273\u0026ndash;282. https://link.springer.com/article/10.1007/s11829-016-9437-9\u003c/li\u003e\n\u003cli\u003eSnoeren TA L, Mumm R, Poelman EH , Yang Y, Pichersky E, Dicke M (2010) The Herbivore-Induced plant volatile methyl salicylate negatively affects attraction of the Parasitoid \u003cem\u003eDiadegma semiclausum\u003c/em\u003e. J Chem Ecol 36:479\u0026ndash;489. https://link.springer.com/article/10.1007/s10886-010-9787-1\u003c/li\u003e\n\u003cli\u003eSteinberg S, Dicke M, Vet LEM (1993) Relative importance of infochemicals from first and second trophic level in long-range host location by the larval parasitoid \u003cem\u003eCotesia glomerata\u003c/em\u003e. J Chem Ecol 19:47\u0026ndash;59. https://link.springer.com/article/10.1007/BF00987470\u003c/li\u003e\n\u003cli\u003eSullivan BT, Pettersson EM, Seltmann KC, Berisford CW (2000) Attraction of the bark beetle parasitoid \u003cem\u003eRoptrocerus xylophagorum\u003c/em\u003e (Hymenoptera: Pteromalidae) to host associated olfactory cues. Environ Entomol 29:1138\u0026ndash;1151.https://doi.org/10.1603/0046-225X-29.6.1138\u003c/li\u003e\n\u003cli\u003eTakemoto H, Takabayashi J (2015) Parasitic wasps \u003cem\u003eAphidius ervi \u003c/em\u003eare more attracted to a blend of host-induced plant volatiles than to the independent compounds. J Chem Ecol\u003cem\u003e \u003c/em\u003e41: 801\u0026ndash;807. https://link.springer.com/article/10.1007/s10886-015-0615-5\u003c/li\u003e\n\u003cli\u003eTam\u0026ograve; C, Ricard I, Held M, Davison AC, Turlings TCJ (2006) A comparison of na\u0026iuml;ve and conditioned responses of three generalist endoparasitoids of lepidopteran larvae to host-induced plant odours. Animal Biol 56:205-220. https://brill.com/view/journals/ab/56/2/article-p205_7.xml\u003c/li\u003e\n\u003cli\u003eThaler JS (1999) Induced resistance in agricultural crops: Effects of jasmonic acid on herbivory and yield in tomato plants. Environ Entomol 28:30-37. https://doi.org/10.1093/ee/28.1.30\u003c/li\u003e\n\u003cli\u003eThaler JS (2002) Effect of jasmonate-induced plant responses on the natural enemies of herbivores. J Anim Ecol\u003cstrong\u003e\u003cem\u003e \u003c/em\u003e\u003c/strong\u003e71:141\u0026ndash;150. https://doi.org/10.1046/j.0021-8790.2001.00586.x\u003c/li\u003e\n\u003cli\u003eThomas C, Maurice SC (2008) Statistix 9, Ninth Edition, Managerial Economics McGraw- Hill /Irwin. (ISBN: 0073402818).\u003c/li\u003e\n\u003cli\u003eTurlings TCJ, Erb M (2018) Tritrophic interactions mediated by herbivore-induced plant volatiles: Mechanisms, ecological relevance, and application potential. Ann Rev Entomol 63:433\u0026ndash;452. https://doi.org/10.1146/annurev-ento-020117-043507\u003c/li\u003e\n\u003cli\u003eTurlings TCJ, Scheepmaker JWA, Vet LEM, Tumlinson JH, Lewis WJ (1990) How contact foraging experiences affect the preferences for host \u0026ndash;related odors in the larval parasitoid \u003cem\u003eCotesia marginiventris\u003c/em\u003e. J Chem Ecol 16:1577-1589. https://link.springer.com/article/10.1007/BF01014091\u003c/li\u003e\n\u003cli\u003eTurlings TCJ, Tumlinson JH, Eller FJ, Lewis WJ (1991) Larval-damaged plants: source of volatile synomes that guide the parasitoid \u003cem\u003eCotesia marginiventris\u003c/em\u003e to the microhabitat of its host. Ent Exp App 58:75-82. https://doi.org/10.1111/j.1570-7458.1991.tb01454.x\u003c/li\u003e\n\u003cli\u003eTurlings TCJ, Alborn HT, Loughrin JH, Tumlinson JH (2000) Volicitin, an elicitor of maize volatiles in oral secretion of \u003cem\u003eSpodoptera exigua\u003c/em\u003e: isolation and bioactivity. J Chem Ecol 26:189-201. https://link.springer.com/article/10.1023/A:1005449730052\u003c/li\u003e\n\u003cli\u003eTurlings TCJ, Davison AC, Tamo C (2004) A six-arm olfactometer permitting simultaneous observation of insect attraction and odour trapping. Physiol Entomol 29:45-55. https://doi.org/10.1111/j.1365-3032.2004.0362.x\u003c/li\u003e\n\u003cli\u003eTurlings TCJ, Jeanbourquin PM, Held M, Degen T (2005) Evaluating the induced-odour emission of a Bt maize and its attractiveness to parasitic wasps. Transg Res 14:807\u0026ndash;816. https://link.springer.com/article/10.1007/s11248-005-0008-6\u003c/li\u003e\n\u003cli\u003eVet LEM, Dicke M (1992) The ecology of infochemical use by natural enemies of herbivores in a tritrophic context. Ann Rev Entomol 37:141\u0026ndash;172.\u003c/li\u003e\n\u003cli\u003eVet LEM, Van Lenteren JC, Heymans M, Meelis E (1983) An airflow olfactometer for measuring olfactory responses of hymenopterous parasitoids and other small insects. Physiol Entomol 8:97\u0026ndash;106. https://doi.org/10.1111/j.1365-3032.1983.tb00338.x\u003c/li\u003e\n\u003cli\u003eWei J, Wang L, Zhao J, Li C, Ge F, Kang L (2011) Ecological tradeoffs between jasmonic acid-dependent direct and indirect plant defences in tritrophic interactions. New Phytolist Trust 189:557\u0026ndash;567. https://nph.onlinelibrary.wiley.com/doi/10.1111/j.1469-8137.2010.03491.x\u003c/li\u003e\n\u003cli\u003eWiedenmann RN, Jr. Smith JW, Darnell PW (1992) Laboratory rearing and biology of the parasite \u003cem\u003eCotesia flavipes\u003c/em\u003e using \u003cem\u003eDiatraea saccharalis\u003c/em\u003e as a host. Environ Entomol 21:1160-1167. https://doi.org/10.1093/ee/21.5.1160\u003c/li\u003e\n\u003cli\u003eWilliams L, Rodriguez-Saona C, Castle del Conte SC (2017) Methyl jasmonate induction of cotton: a field test of the \u0026lsquo;attract and reward\u0026rsquo; strategy of conservation biological control. AoB PLANTS, 9 (5), plx032. https://doi.org/10.1093/aobpla/plx032\u003c/li\u003e\n\u003cli\u003eXiu C, Dai W, Pan H et al (2019) Herbivore-induced plant volatiles enhance field-level parasitism of the mirid bug \u003cem\u003eApolygus lucorum\u003c/em\u003e. Biol Control 135:41\u0026ndash;47. https://doi.org/10.1016/j.biocontrol.2019.05.004\u003c/li\u003e\n\u003cli\u003eYe M, Song Y, Long J, Wang R, Baerson SR, Pan Z, et al. (2013) Priming of jasmonate-mediated antiherbivore defense responses in rice by silicon. PNAS- USA, 110:E3631\u0026ndash;E3639. \u003cu\u003edoi: 10.1073/pnas.1305848110\u003c/u\u003e.\u003c/li\u003e\n\u003cli\u003eYe M, Veyrat N, Xu H, Hu L, Turlings TCJ, Erb M (2018) An herbivore-induced plant volatile reduces parasitoid attraction by changing the smell of caterpillars. Sci. Adv. 4, eaar4767. https://www.science.org/doi/full/10.1126/sciadv.aar4767\u003c/li\u003e\n\u003cli\u003eZar JH: Biostatistical analysis. Fourth edition, Prentice Hall, Upper Saddle River, New Jersey, USA, 663 pp (1999).\u003c/li\u003e\n\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":"Cotesia marginiventris, Olfactometer, Jasmonate, Cotton, Beet armyworm","lastPublishedDoi":"10.21203/rs.3.rs-5306734/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-5306734/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eIt is well documented that plants damaged with insect species can attract specific biocontrol agents throughout the emission of insect-induced volatiles. A four-armed olfactometer was used to inspect olfactory reactions of \u003cem\u003eCotesia marginiventris\u003c/em\u003e. Behavioural tests for studying parasitoid attraction to specific odours are assessed. Flying tunnel investigations could be evaluated with three odour sources plus control plants at a time. The attractiveness of female parasitoid \u003cem\u003eC. marginiventris\u003c/em\u003e was tested by applying several treatments: jasmonate induced plants and plants fed by \u003cem\u003eSpodoptera exigua\u003c/em\u003e on \u003cem\u003eBacillus thuringiensis\u003c/em\u003e (BT)(genetic modified GM) and non- GM cotton plants. Three criteria were compared and studied. Search time and choose time and visit numbers for each zone.\u003c/p\u003e\n\u003cp\u003eThe obtained outcomes showed that the plants induced with jasmonate were more attractive than those plants fed by two larvae, while those plants fed by one larva were the least attractive. Whereas, it was found that females of \u003cem\u003eC. marginiventris \u003c/em\u003esignificantly chose more jasmonate feeding areas than larval feeding areas on GM cotton plants, and the equivalent effect was detected on non-GM cotton plants. Whereas \u003cem\u003eC.\u003c/em\u003e \u003cem\u003emarginiventris\u003c/em\u003e females spent more time in the jasmonate areas than the rest of the treatments; While the time spent on transgenic cotton plants was longer than the time spent on non-transgenic cotton plants. Whereas in the non-Jasmonate treatments, females spent more time using the GM cotton than the non-GM cotton, the same trend of effects had been detected in the choice of time and visit frequency for that treatment.\u003c/p\u003e\n\u003cp\u003eJasmonate stimulation of volatile substances is a good way to increase the attractiveness of parasitoids to their insect hosts to maximize the efficacy of the role of biological control agents. It is proposed that a genetic modification approach to modify the volatiles in these plants could enhance the attractiveness and increase the role assigned to natural enemies. This procedure will cause a more drastic change in the volatiles of those plants. This scientific direction is a complementary role, with transgenic resistance, which will lead to selection of inducible promoters with tissue-specific expression.\u003c/p\u003e","manuscriptTitle":"Effectiveness of jasmonate stimulation on attraction of Cotesia marginiventris via olfactory cues on BT- and non-BT-cotton plants","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-11-14 16:36:10","doi":"10.21203/rs.3.rs-5306734/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":"67a4ca58-5cfa-43e3-8423-3c8b7785ccc2","owner":[],"postedDate":"November 14th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-02-03T05:26:41+00:00","versionOfRecord":[],"versionCreatedAt":"2024-11-14 16:36:10","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-5306734","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-5306734","identity":"rs-5306734","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}