CRISPR/Cas9 mediated validation of spermatogenesis-related gene, tssk2 as a component of genetic pest management of fall armyworm, Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae).

Invasive insect pests, currently, pose a serious economic threat to several staple crops all over the world, one such being the fall armyworm, Spodoptera frugiperda. It was first observed in Africa since 2016, outside of its natural habitat in the Americas. Subsequently, it invaded several countries in South and South East Asia and also very recently in Australia. In all the newly invaded regions, maize is the principal crop attacked causing a serious economic concern to the poor farmers, particularly in the developing countries. Owing to the innate genetic ability, it defies many of the management options that include insecticides, Bt transgenics, and so forth. This is due to its high mobility, polyphagy and ability for quick development of resistance to several classes of insecticides. At this critical juncture, CRISPR/Cas9 mediated genome editing has shown a lot of promise in developing a novel area-wide pest management strategy called precision-guided sterile insect technique (pgSIT). pgSIT was initially demonstrated in Drosophila melanogaster which holds a greater promise for the environmentally friendly management of several globally significant agricultural pests such as S. frugiperda. Therefore, before developing both sgRNA and Cas9 transgenic lines, we have validated the target gene such as tssk2 through a non-transgenic approach by microinjecting ribo nucleo protein complex (Cas9 protein and tssk2 sgRNA) into G0 eggs of S. frugiperda. In the current investigation, we have obtained five edited males with distinct mutations which were further used for crossing studies to ascertain the effect of tssk2 editing affecting egg hatchability.

Efficacy of conventional and organic pesticides following ingestion by Delia radicum (Diptera: Anthomyiidae).

Cabbage maggot (CM) (Delia radicum L.) is a devastating pest of Brassicaceae crops throughout the world, including the Willamette Valley in western Oregon, USA. Chemical control methods for this pest are limited, with reduction or elimination of chlorpyrifos tolerances and expensive alternative chemistries; therefore, there is an increasing need for novel chemical control options. Adult feeding, a strategy used with insecticide-treated baits for other fly species, has yet to be tested as an option for a chemical control delivery for cabbage maggot. Treated bait can exploit the feeding behavior of CM and expose them to insecticides in a field setting. In this study, the efficacy of 5 organic and 5 conventional insecticides was compared in laboratory bioassays of treated bait stations in Aurora, Oregon, USA. The mortality of adult female cabbage maggot flies was assessed over time following ingestion of insecticides. Among organic insecticides tested, spinosad was highly effective 4 h after exposure, while pyrethrins + azadirachtin was moderately effective following 18 h after exposure. Flies exposed to conventional-use pesticides zeta-cypermethrin and bifenthrin had high mortality 1.75 h after exposure, while spinetoram had moderate efficacy 2 h after exposure. Insecticides identified with high or moderate efficacy may have the potential for use in baits or lure formulations that could be used to augment the control of cabbage maggots in field settings.

Image detection model construction of Apolygus lucorum and Empoasca spp. based on improved YOLOv5.

BACKGROUND: The polyphagous mirid bug Apolygus lucorum (Meyer-Dür) and the green leafhopper Empoasca spp. Walsh are small pests that are widely distributed and important pests of many economically important crops, especially kiwis. Conventional monitoring methods are expensive, laborious and error-prone. Currently, deep learning methods are ineffective at recognizing them. This study proposes a new deep-learning-based YOLOv5s_HSSE model to automatically detect and count them on sticky card traps. RESULTS: Based on a database of 1502 images, all images were collected from kiwi orchards at multiple locations and times. We trained the YOLOv5s model to detect and count them and then changed the activation function to Hard swish in YOLOv5s, introduced the SIoU Loss function, and added the squeeze-and-excitation attention mechanism to form a new YOLOv5s_HSSE model. Mean average precision of this model in the test dataset was 95.9%, the recall rate was 93.9% and the frames per second was 155, which are higher than those of other single-stage deep-learning models, such as SSD, YOLOv3 and YOLOv4. CONCLUSION: The proposed YOLOv5s_HSSE model can be used to identify and count A. lucorum and Empoasca spp., and it is a new, efficient and accurate monitoring method. Pest detection will benefit from the broader applications of deep learning. © 2024 Society of Chemical Industry.

Potential of Ocotea indecora Essential Oil for Controlling Drosophila suzukii: Molecular Predictions for Toxicity and Selectivity to Beneficial Arthropods.

The protection of soft-skinned fruits against Drosophila suzukii has relied primarily on the efficacy of a few synthetic molecules. Despite their short-term efficacy, these molecules can cause environmental pollution, unintendedly affect non-target organisms, and fail to provide sustainable control. The shortfalls of using synthetic pesticides warrant the search for alternatives, such as essential oils extracted from plants, with greater eco-friendlier properties. Here, we chemically characterized and evaluated the toxicity of the essential oil extracted from leaves of Ocotea indecora (Schott) Mez (Lauraceae) against D. suzukii via two exposure pathways (ingestion and contact). We also assessed the selectivity of the essential oil to two predatory natural enemies, Eriopis connexa and Chrysoperla externa and two pollinator bees, Apis mellifera and Partamona helleri. In addition, we conducted in silico predictions to investigate potential interactions between the major compound of the essential oil and the insects' transient receptor potential (TRP) channels. Our chromatographic analysis revealed sesquirosefuran (87%) as the major compound. Higher toxicity to adults of D. suzukii was observed in contact exposure (LC50 = 0.43 µL mL-1) compared to ingestion (LC50 = 0.72 µL mL-1). However, the essential oil did not cause mortality to the non-target organisms tested here, even when applied at 2.20 µL mL-1. Molecular predictions demonstrated that sesquirosefuran binds more stably to the TRP channels of D. suzukii than to those expressed in beneficial arthropods. Collectively, our findings provide the initial framework for the potential use of O. indecora essential oil as a sustainable alternative for managing D. suzukii infestations.

Natural Oil Lure Outperforms Trimedlure in Capturing Males of the Mediterranean Fruit Fly, Ceratitis capitata (Diptera: Tephritidae).

Females of certain tephritid fruit fly species (Diptera: Tephritidae) pose an enormous agricultural threat, as they oviposit in commercially important fruits and vegetables. Trapping networks are often operated in fruit fly-free areas to detect incipient infestations. Trapping relies largely on male attractants, so-called male lures, with trimedlure (TML) being used to detect invasive Ceratitis spp. Operating large-scale surveillance programs incurs substantial costs for both supplies and labor, and the problem is exacerbated by the fact that trimedlure (as well as other male lures) is effective for relatively short intervals in the field (6-8 weeks). Because frequent servicing increases costs, there is considerable interest in modifying existing lures or developing new formulations to extend their effective field longevity. Here, we present results of a field study in Hawaii on a wild population of the Mediterranean fruit fly, Ceratitis capitata (Wiedemann), that compared male captures in traps baited with (i) fresh liquid TML, (ii) TML plugs, (iii) a novel controlled-release TML sachet, and (iv) a novel natural oil blend dispensed from a sachet. Catch was recorded weekly for 12 weeks and then at 16 and 20 weeks, with 12 traps deployed per treatment. The natural oil formulation, which contains the natural plant product α-copaene, was as effective as the fresh liquid TML even after weathering for 20 weeks. Future work will focus on developing a dispenser for this formulation that is compatible with standard trap design and deployment.

Comparison of different traps and attractants in 3 food processing facilities in Greece on the capture of stored product insects.

Certain lures are marketed toward particular pests or classes of pests, while others might be multi-species lures. Investigative aims for this study included both which trap was most sensitive and whether different combinations of traps and attractants were delivering novel information about the stored product insect community. Comparisons were made for all combinations of 3 commercial traps and 4 different attractants plus an untreated control on the capture of stored-product insects for 2 consecutive years in 3 food processing facilities in Central Greece. The traps used in the experiments were Dome Trap (Trécé Inc., USA), Wall Trap (Trécé) and Box Trap (Insects Limited, Ltd., USA). The attractants that were evaluated were 0.13 g of (i) PantryPatrol gel (Insects Limited), (ii) Storgard kairomone food attractant oil (Trécé), (iii) wheat germ (Honeyville, USA), and (iv) Dermestid tablet attractant (Insects Limited). The traps were inspected approximately every 15 days and rotated. A total of 34,000+ individuals were captured belonging to 26 families and at least 48 species. The results indicated that Indian meal moth, Plodia interpunctella (Hübner), red flour beetle, Tribolium castaneum (Herbst), and cigarette beetle, Lasioderma serricorne (F.) were the most abundant. Although there were noticeable differences among the different traps and attractants for specific species, all combinations provided similar information on population dynamics. Generally, Dome traps baited with either the oil or the gel, were found to be the most sensitive. The results of the present study demonstrate the importance of long-term trapping protocols, as a keystone in IPM-based control strategies in food processing facilities.

Gas exchange patterns for a small, stored-grain insect pest, Tribolium castaneum.

Insects breathe using one or a combination of three gas exchange patterns; continuous, cyclic and discontinuous, which vary in their rates of exchange of oxygen, carbon dioxide and water. In general, there is a trade-off between lowering gas exchange using discontinuous exchange that limits water loss at the cost of lower metabolic rate. These patterns and hypotheses for the evolution of discontinuous exchange have been examined for relatively large insects (>20 mg) over relatively short periods (<4 h), but smaller insects and longer time periods have yet to be examined. We measured gas exchange patterns and metabolic rates for adults of a small insect pest of grain, the red flour beetle, Tribolium castaneum (Coleoptera: Tenebrionidae), using flow-through respirometry in dry air for 48 h. All adults survived the desiccating measurement period; initially they used continuous gas exchange, then after 24 h switched to cyclic gas exchange with a 27% decrease in metabolic rate, and then after 48 h switched to discontinuous gas exchange with increased interburst duration and further decrease in metabolic rate. The successful use of the Qubit, a lower cost and so more common gas analyser, to measure respiration in the very small T. castaneum, may prompt more flow-through respirometry studies of small insects. Running such studies over long durations may help to better understand the evolution of respiration physiology and thus suggest new methods of pest management.

Effects of Color Attributes on Trap Capture Rates of Chrysobothris femorata (Coleoptera: Buprestidae) and Related Species.

Chrysobothris spp. (Coleoptera: Buprestidae) and other closely related buprestids are common pests of fruit, shade, and nut trees in the United States. Many Chrysobothris spp., including Chrysobothris femorata, are polyphagous herbivores. Their wide host range leads to the destruction of numerous tree species in nurseries and orchards. Although problems caused by Chrysobothris are well known, there are no reliable monitoring methods to estimate local populations before substantial damage occurs. Other buprestid populations have been effectively estimated using colored sticky traps to capture beetles. However, the attraction of Chrysobothris to specific color attributes has not been directly assessed. A multi-color trapping system was utilized to determine color attraction of Chrysobothris spp. Specific color attributes (lightness [L*], red to green [a*], blue to yellow [b*], chroma [C*], hue [h*], and peak reflectance [PR]) were then evaluated to determine beetle responses. In initial experiments with mostly primary colors, Chrysobothris were most attracted to traps with red coloration. Thus, additional experiments were performed using a range of trap colors with red reflectance values. Among these red reflectance colors, it was determined that the violet range of the electromagnetic spectrum had greater attractance to Chrysobothris. Additionally, Chrysobothris attraction correlated with hue and b*, suggesting a preference for traps with hues between red to blue. However, males and females of some Chrysobothris species showed differentiated responses. These findings provide information on visual stimulants that can be used in Chrysobothris trapping and management. Furthermore, this information can be used in conjunction with ecological theory to understand host-location methods of Chrysobothris.

Drosophila suzukii Management in Latin America: Current Status and Perspectives.

Spotted-wing drosophila, Drosophila suzukii Matsumura, was first established in Latin America in Mexico in 2011. The vinegar fly has since been detected in 296 municipalities in Argentina, Brazil, Chile, Mexico, and Uruguay. Drosophila suzukii is polyphagous and is found on 64 host plants in 25 families in Latin America, with most hosts also exotic species. In Latin America, D. suzukii is attacked by 14 species of parasitoid wasps in the families Diapriidae, Figitidae, and Pteromalidae, which are promising native parasitoids for control of the pest. This article analyzes results from studies on monitoring, biological, chemical, and cultural control, and sterile insect techniques to provide a basis for the development of area-wide and sustainable D. suzukii management programs in Latin America. The review examines how D. suzukii has been managed in Latin America and how research conducted in this region can contribute to management of the species in other parts of the world.

Prevalence of Cockroaches, Bed Bugs, and House Mice in Low-Income Housing and Evaluation of Baits for Monitoring House Mouse Infestations.

Pests are frequently found in homes, especially in apartment buildings in low-income communities. We investigated the prevalence and patterns of pest infestations in low-income communities in four cities (Jersey City, Linden, Paterson, Trenton) in New Jersey, USA. Resident interviews, visual inspections, and the placement of monitors were used to identify pest infestations. A total of 1,753 apartments from 19 buildings or building complexes were accessed. The infestation rates of cockroaches, bed bugs (Cimex lectularius L.), and house mice (Mus musculus domesticus Schwarz and Schwarz) were 37, 9, and 20%, respectively. Among apartments with cockroaches, 97.8, 2.5, and 0.8% had German cockroach [Blattella germanica (L.)], American cockroach [Periplaneta americana (L.)], and Oriental cockroach (Blatta orientalis L.), respectively. The percentage of residents who were aware of the presence of cockroaches, bed bugs, and house mice was 70.8, 55.3, and 56.8%, respectively. The prevalence of pest infestation was associated with resident ethnicity and gender. Among 856 interview responses, 78% implemented methods themselves to control pests in their homes in the past six months. Chocolate spread detected 99% of house mouse infestations and was much more sensitive than three commercial blank baits. Significant differences were also observed in the feeding preference of the three commercial blank baits.

Insight into the evolutionary profile of radio-resistance among insects having intrinsically evolved defence against radiation toxicity.

Ionizing radiation (IR) has wide-ranging applications in various fields. In agriculture, pest control is one of the important applications, because insect pests have become a threat to the global agriculture industry. IR are used routinely to prevent crop loss and to protect stored food commodities. Radio-sterilization and disinfestation treatments are commonly used procedures for insect pest control. From various studies on insect radio-sterilization and disinfestation, it has been established that compared to vertebrates' insects have high levels of radiation resistance. Therefore, to achieve adequate radio-sterilization/disinfestation; exposure to high doses of IR is necessary. However, studies over decades made a presumption that radiation resistance is general among insects. Recent studies have shown that some insect orders are having high IR resistance and some insect orders are sensitive to IR. These studies have clarified that radiation resistance is not uniform throughout insect class. The present review is an attempt to insight at the evolutionary profile of insect species studied for radio-sterilization and disinfestation treatment and are having the trait of radio-resistance. From various studies on insect radiation resistance and after phylogenetic analysis of insect species it appears that the evolutionary near species have drastically different levels of radio-resistance and trait of radiation resistance appears to be independent of insect evolution.

Liquid preparation of diatomaceous earth against the lesser mealworm, Alphitobius diaperinus (Panzer, 1797) (Coleoptera: Tenebrionidae)

The lesser mealworm (LMW) is the most important pest in poultry production. Insects are associated to avian pathogens, mainly Salmonella. Its control is based on chemical insecticide, with limited efficacy in population reduction. Also, insect populations are resistant to the main active ingredients used. Diatomaceous earth (DE) is a mineral dust that has activity against the LMW when used in dust application. No information is available about DE liquid preparation against this insect. Thus, bioassays were conducted aiming to develop a strategy for DE liquid preparation to this insect control. In laboratory the ideal concentration for poultry house experiment was estimated and the effect of insect contact with poultry litter or chicken feed after application was checked. In the poultry house, DE liquid preparation (10% in water, 1 L·m­2) was applied on the dirt soil of a cleaned and empty poultry house. In the control poultry house, a chemical insecticide was used (cypermethrin 15 g + chlorpyriphos 25 g + piperonyl butoxide 15 g) (1 L·m­2). DE at all concentrations was efficient mainly when insects were dried for 24 h before contact with a substrate. In poultry house, both treatments obtained similar results at 15 days after treatment (94% of reduction of insect population). At 45 days after treatment, the insect population with DE treatment increased 39% while the insect population remained 17% smaller than the initial population with a chemical insecticide. DE liquid preparation has potential to be used as a safe treatment in LMW population management, as a shock treatment.

60Co-γ Radiation Alters Developmental Stages of Zeugodacus cucurbitae (Diptera: Tephritidae) Through Apoptosis Pathways Gene Expression.

Radiation is considered as a promising insect pest control strategy for minimizing postharvest yield losses. Among various techniques, irradiation is a method of choice as it induces lethal biochemical or molecular changes that cause a downstream cascade of abrupt physiological abnormalities at the cellular level. In this study, we evaluated the effect of 60Co-γ radiation on various developmental stages of Zeugodacus cucurbitae Coquillett and subsequent carry-over effects on the progeny. For this purpose, we treated eggs with 30- and 50-Gy radiation doses of 60Co-γ. We found that radiation significantly affected cellular antioxidants, insect morphology, and gene expression profiles. Our results indicate that in response to various doses of irradiation reactive oxygen species, catalase, peroxidase, and superoxide dismutase activities were increased along with a significant increase in the malondialdehyde (MDA) content. We observed higher mortality rates during the pupal stage of the insects that hatched from irradiated eggs (50 Gy). Furthermore, the life span of the adults was reduced in response to 50 Gy radiation. The negative effects carried over to the next generation were marked by significantly lower fecundity in the F1 generation of the irradiation groups as compared to control. The radiation induced morphological abnormalities at the pupal, as well as the adult, stages. Furthermore, variations in the gene expression following irradiation are discussed. Taken together, our results signify the utility of 60Co-γ radiation for fruit fly postharvest management.

Pesticides Misused for Bed Bug Control: Comparing Professional and Nonprofessional Applications Reported to the National Pesticide Information Center, 2013-2017.

Objectives. To compare outcomes when pesticides are used to control bed bugs by professionals and nonprofessionals. Methods. All US National Pesticide Information Center inquiries from 2013 to 2017 were assessed to identify scenarios involving bed bugs and pesticide applications. Cases were evaluated with respect to types of applicators, misapplications, and human pesticide exposures. Results. Misapplications were more than twice as likely to be reported in cases involving nonprofessional applications (14%) as in cases involving professional applications (5%). Human exposures to pesticides were reported more often when pesticides were misapplied (70%) than when there were no apparent misapplications (31%). Conclusions. Both professionals and nonprofessionals may misuse pesticides to control bed bugs, which may increase the risks of exposure and adverse outcomes. Policy interventions may reduce pesticide incidents related to bed bug control by promoting professional involvement and adherence to product label instructions.

Pest and disease management by red light.

Light is essential for plant life. It provides a source of energy through photosynthesis and regulates plant growth and development and other cellular processes, such as by controlling the endogenous circadian clock. Light intensity, quality, duration and timing are all important determinants of plant responses, especially to biotic stress. Red light can positively influence plant defence mechanisms against different pathogens, but the molecular mechanism behind this phenomenon is not fully understood. Therefore, we reviewed the impact of red light on plant biotic stress responses against viruses, bacteria, fungi and nematodes, with a focus on the physiological effects of red light treatment and hormonal crosstalk under biotic stress in plants. We found evidence suggesting that exposing plants to red light increases levels of salicylic acid (SA) and induces SA signalling mediating the production of reactive oxygen species, with substantial differences between species and plant organs. Such changes in SA levels could be vital for plants to survive infections. Therefore, the application of red light provides a multidimensional aspect to developing innovative and environmentally friendly approaches to plant and crop disease management.

DNA screening of Drosophila suzukii predators in berry field orchards shows new predatory taxonomical groups.

Drosophila suzukii (spotted wing drosophila, SWD) is a pandemic quarantine pest that attacks mostly red fruits. The high number of life cycles per year, its ability to rapidly invade and spread across new habitats, and highly polyphagous nature, makes this a particularly aggressive invasive species, for which efficient control methods are currently lacking. The use of native natural predators is particularly promising to anchor sustainable and efficient measures to control SWD. While several field studies have suggested the presence of potential predatory species in infested orchards, only a few confirmed the presence of SWD DNA in predators' gut content. Here, we use a DNA-based approach to identify SWD predators among the arthropod diversity in South Europe, by examining the gut content of potential predator specimens collected in SWD-infested berry fields in North Portugal. These specimens were morphologically identified to the family/order, and their gut content was screened for the presence of SWD DNA using PCR. New SWD predatory taxonomical groups were identified, as Opiliones and Hemerobiidae, in addition to known SWD predators, such as Hemerobiidae, Chrysopidae, Miridae, Carabidae, Formicidae and Araneae. Additionally, the presence of a spider family, Uloboridae, in the orchards was recorded for the first time, posing this family as another SWD-candidate predator. This study sets important bases to further investigate the potential large-scale use of some of these confirmed predator taxa for SWD control in South Europe.

Induced Hatching of Quiescent Aedes aegypti (Diptera: Culicidae) Eggs by Labile Glutathione-Stabilizable Compounds From Yeast Extract.

Aedes aegypti (Linnaeus, 1762) is the insect vector that transmits several deadly human diseases. Although the egg stage is an important phase of its life cycle, the biology of mosquito egg remains poorly understood. Here, we report our investigations on the chemical factors that induced hatching of Ae. aegypti eggs. Commercial yeast extracts were able to increase egg hatching rate in a dose-dependent manner, with a hatching rate that ranged from approximately 10% with 1 g/liter to 80% with 20 g/liter of yeast extract. Notably, the addition of glutathione, a reducing agent that showed no significant effect on egg hatching by itself, enhanced and stabilized the activity of yeast extract for at least 70 h. Because dissolved oxygen in different treatments was maintained at high levels in a narrow range (92-95%), we proposed that yeast extract contains hatching inducing compound (HIC) which is able to trigger egg hatching independent of dissolved oxygen level. The HIC in yeast extract could prove to be a potential starting point to design an effective tool to forcefully induce mosquito eggs to hatch under unfavorable conditions, functioning as a novel method for vector control.

Biotechnological interventions for the sustainable management of a global pest, whitefly (Bemisia tabaci).

Whiteflies (Bemisia tabaci) are polyphagous invasive hemipteran insects that cause serious losses of important crops by directly feeding on phloem sap and transmitting pathogenic viruses. These insects have emerged as a major threat to global agriculture and food security. Chemically synthesized insecticides are currently the only option to control whiteflies, but the ability of whiteflies to evolve resistance against insecticides has made the management of these insects very difficult. Natural host-plant resistance against whiteflies identified in some crop plants has not been exploited to a great extent. Genetic engineering approaches, such as transgenics and RNA interference (RNAi), are potentially useful for the control of whiteflies. Transgenic plants harboring insecticidal toxins/lectins developed via nuclear or chloroplast transformation are a promising vehicle for whitefly control. Double-stranded RNAs (dsRNAs) of several insect genes, delivered either through microinjection into the insect body cavity or orally via an artificial diet and transiently or stably expressed in transgenic plants, have controlled whiteflies in model plants and in some crops at the laboratory level, but not at the field level. In this review, we highlight the merits and demerits of each delivery method along with strategies for sustained delivery of dsRNAs via fungal entomopathogen/endosymbiont or nontransgenic RNAi approaches, foliar sprays, root absorption or nanocarriers as well as the factors affecting efficient RNAi and their biosafety issues. Genome sequencing and transcriptome studies of whitefly species are facilitating the selection of appropriate genes for RNAi and gene-editing technology for the efficient and resilient management of whiteflies and their transmitted viruses.

Volatile Organic Compounds as Insect Repellents and Plant Elicitors: an Integrated Pest Management (IPM) Strategy for Glasshouse Whitefly (Trialeurodes vaporariorum).

The glasshouse whitefly (Trialeurodes vaporariorum Westwood) is a polyphagous arthropod pest that is of particular detriment to glasshouse grown tomato (Solanum lycopersicum) across temperate regions of the world. Control of whiteflies with synthetic pesticides has resulted in the evolution of resistant genotypes and a reduction in natural enemies, thus highlighting the need for environmentally sound control strategies. Volatile organic compounds (VOCs) offer an environmentally benign alternative to synthetic chemical sprays and this study explored the use of VOCs as insect repellents and plant defence elicitors to control whiteflies on tomato in a commercial glasshouse setting. Limonene in the form of a volatile dispenser system was found to successfully repel whitefly from the target crop and increased fruit yield by 32% during a heavy whitefly infestation. Analysis of tomato herbivore induced plant volatiles (HIPVs) led us to select methyl salicylate (MeSA) as the plant elicitor and application of MeSA to un-infested tomato plants was found to successfully reduce whitefly population development and increase yield by 11%, although this difference was marginally statistically significant. Combination of these two methods was also effective but whitefly abundance in combined plots was similar to the standalone limonene treatment across the course of the experiment. All of the VOC based control methods we used had a negative impact on whitefly performance, with more pronounced effects during the first few weeks of infestation. In subsequent laboratory experiments, we found elevated peroxidase (POD) activity and a significant increase in TPX1 and PR1 transcripts in MeSA treated plants. This led us to deduce that MeSA immediately induced plant defences, rather than priming them. We did however see evidence for residual priming, as plants treated with MeSA and infested with whiteflies produced significantly higher levels of POD activity than whitefly infestation alone. Despite the fact that our treatments failed to synergise, our methods can be optimised further, and the effectiveness of the standalone treatments is promising for future studies. In particular, our repellent limonene dispensers were extremely effective at deterring whiteflies and offer a low economic cost and easy to implement whitefly control option. The methods we have used here could be incorporated into current integrated pest management (IPM) systems, a sustainable approach to pest control which will be central to our efforts to manage whitefly populations under glass in the future.