Persistence of widespread moderate Spinosad resistance among wild melon fly (Zeugodacus cucurbitae) and oriental fruit fly (Bactrocera dorsalis) populations on the major Hawaiian islands.

BACKGROUND: Insecticide resistance among invasive tephritid fruit flies poses a great risk to national food security and has the potential to disrupt quarantine and eradication programs, which rely on the efficacy of Spinosad to prevent widespread establishment in North America. During 2022 to 2023 we surveyed the extent of Spinosad resistance of two key species, oriental fruit fly Bactrocera dorsalis, and melon fly Zeugodacus cucurbitae, from 20 sites across five Hawaiian Islands including Kaua'i, O'ahu, Maui, Molokai and the "Big Island" (Hawai'i). RESULTS: We used topical thoracic applications of eight concentrations of Spinosad ranging from 0.028 to 3.6 mg/mL to evaluate the lethal concentration (LC50 and LC99) required to kill wild-caught males. Resistance ratios (RR) were calculated by comparing the LC50 of wild flies to laboratory susceptible lines maintained in colony. Our results identified at least two new sites of concern for melon fly resistance on the Big Island, and at least four sites of concern for oriental fruit fly, all of which were located on the Big Island. At these locations RRs were >5. On O'ahu, melon fly RRs were >10. CONCLUSIONS: The persistence of Spinosad resistance is concerning, yet it is a reduction compared to the values reported previously and before changes to Spinosad use recommendations by local extension agents beginning in 2017. For oriental fruit fly, these RR values are the highest levels that have been detected in wild Hawai'i populations. These data suggest that expanded Spinosad reduction and replacement programs are warranted given the ongoing issues with Spinosad resistance in Hawai'i and expansion in the number of species affected. © 2024 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

Avocado cultivar and tree-to-tree leaf compositional differences affect infestation severity of Pseudocysta perseae (Hemiptera: Tingidae).

Avocado lace bug, Pseudocysta perseae (Heidemann) (Hemiptera: Tingidae), is a sap-feeding insect that feeds on the underside of avocado leaves. First observed in 2019, P. perseae has spread throughout the Hawaiian islands, causing premature leaf drop and decrease in avocado yield. Due to Hawai'i's approximately 200 cultivars comprised of all 3 avocado races with extensive racial hybrids, we were able to investigate whether certain cultivars were more prone to experiencing higher P. perseae abundances and infestations compared to others. We conducted longitudinal abundance surveys on Hawai'i Island across several common avocado varieties monitoring changes in P. perseae abundance. These surveys were supplemented with longitudinal infestation severity surveys across 4 avocado lineages (Mexican, Guatemalan, West Indian, and Guatemalan × West Indian hybrid). Additionally, we collected leaves of 'Sharwil', 'Hass', 'Kahalu'u', and 'Nishikawa' cultivars looking at associations between P. perseae abundance and cultivar, herbivory-related biomechanical traits, and soluble sugar content. We found that some cultivars, such as 'Malama', typically experience lower P. perseae abundances compared to cultivars such as 'Kahalu'u', 'Beshore', and 'Sharwil'. Guatemalan × West Indian hybrid trees were also shown to have a higher probability of experiencing more severe P. perseae infestations compared to other lineages. Lastly, soluble sugar content, specifically fructose content, had a positive effect on juvenile P. perseae abundance. These findings suggest that cultivar differences in P. perseae infestations may exist, but tree-to-tree leaf compositional differences, such as soluble sugar content, may be a large driver of variation in P. perseae abundance.

Understanding the movement and dispersal patterns of released Fopius arisanus (Hymenoptera: Braconidae) parasitoids in a papaya orchard.

Implementation of augmentative biological control requires estimates of parasitoid dispersal from the release point to determine appropriate release density, spacing, and timing. This study evaluated the movement patterns of Fopius arisanus Sonan (Hymenoptera: Braconidae) parasitoids, which have historically been used to control invasive tephritid fruit flies. The wasps were released from the central point, and dispersal was monitored over time using parasitism in sentinel fruit and trap captures at 40 points radiating out from the center (15-240 m). The releases were conducted 4 times during June, July, September, and November 2006. The data showed that there were large declines in dispersal by distance. Parasitism was greatest closest to the release point, within 30 m. Parasitism was also greatest within the first 24 h of the release. After 1 wk, parasitism decreased from 41% to 1.5% within 30 m. These data correlated strongly with trap capture data, which also showed that parasitoid movement favored the SE region of our release site, roughly corresponding to the overall prevailing winds. Wind speed, relative humidity, and temperature all affected parasitoid movement during our trial, indicating the complex environmental factors that can affect release success. This is the first report of dispersal metrics for F. arisanus. Our findings are in agreement with other similar studies on braconid movement generally and suggest that frequent, high-density releases are most effective since the dispersal of F. arisanus is limited and retention in the environment is low. We discuss our results in the context of international augmentative biological control release programs.

Previously introduced braconid parasitoids target recent olive fruit fly (Bactrocera oleae) invaders in Hawai'i.

The olive fruit fly Bactrocera oleae (Diptera: Tephritidae) was detected on Maui and Hawai'i Islands in 2019, affecting yields and quality of the state's emerging olive oil industry. Given previous parasitoid releases to control other invasive frugivorous tephritids in Hawai'i, we were interested in determining whether these parasitoids were naturally targeting recent olive fly invaders in field, if local olive cultivar differences affected parasitization rates, and if there was a seasonal pattern of parasitization that could inform future management decisions. To address these questions, we collected data from olive growing in Hawai'i during 2021 and 2022. During the fruiting season we collected monthly samples and reared out B. oleae in the lab. We detected two previously introduced braconid wasps: first Diachasmimorpha tryoni during 2021 and 2022 and later Fopius arisanus during the 2022 collection. Cultivar effects were limited to a single site in our study, where more D. tryoni were reared from 'Arbequina' olives. Seasonality of olive fruit fly and parasitoid activity was earlier in lower elevation sites, as expected based on tree phenology and temperature-dependent insect development. This represents the first report of D. tryoni parasitism activity against B. oleae and may reflect elevational effects combined with the ecological complexity in interactions between multiple invasive arthropod pests, their invasive and cultivated plant hosts, and introduced braconid parasitoids.

Physical, Chemical, and Sensory Properties of a Turmeric-Fortified Pineapple Juice Beverage.

Beverage mixtures based on pineapple (Ananas comosus) and turmeric (Curcuma longa) juice as a ready-to-drink product were developed, and their physicochemical, nutritional, and sensory properties were evaluated. Four different concentrations of turmeric juice (5%, 10%, 15%, and 20% (v/v)) were added to pineapple juice to make turmeric-fortified pineapple (TFP) juice samples. Pineapple juice without turmeric was the control. The L*, a*, b*, titratable acidity (TA), total antioxidant capacity, and %DPPH scavenging values, as well as the concentrations of the phenolic compounds curcumin and demethoxycurcumin, were significantly increased with increasing turmeric concentration. Thirty volatile compounds were detected in the mixed juice samples with turmeric. Most of the turmeric-specific compounds, including monoterpenes, sesquiterpenes and turmerones, were detected in the TFP juice samples. While the antioxidant activity of the juice samples increased with increasing turmeric concentration, the pineapple juice fortified with 10% turmeric (10%T) had the best overall quality as determined by panelists. Greater concentrations of turmeric were associated with decreased palatability due to reduced mouthfeel and sweetness and increased aftertaste and sourness. These results suggest that the 10%T juice could be developed into a commercial functional beverage with increased overall flavor and nutritional quality.

Susceptibility of 'Malama' Avocados to Bactrocera dorsalis (Diptera: Tephritidae) in Hawai'i and a Proposed Systems Approach for Market Access.

Avocados are generally poor hosts or nonhosts for tephritid fruit flies, which can influence the type of mitigation measures required by export protocols for trade. We conducted experiments to determine the susceptibility of the avocado, Persea americana, cv. 'Malama' to infestation by Oriental fruit fly, Bactrocera dorsalis (Hendel) (Diptera: Tephritidae), in Hawai'i using laboratory and field no-choice cage tests, together with field sampling. In no-choice cage tests, infestation rates increased with decreasing fruit firmness over time after harvest. Although risk of infestation was negatively correlated with fruit firmness, about 50% of mature hard fruit exposed immediately after harvest became infested. In field cage tests, mature hard fruit on the tree at the 25% purple color stage were not infested, whereas fruit at the 50, 75, and 100% purple color stages were infested at relatively low rates. Field sampling of fallen 'Malama' fruit over two seasons showed no natural infestation by B. dorsalis and trap captures in avocado orchards using McPhail traps containing protein bait lures averaged 0.13 ± 0.03 (mean ± SE) flies per trap per week, underscoring the low prevalence of fruit flies in avocado orchards and the poor host quality of this cultivar. A systems approach based on poor host status, low prevalence, and a limited harvest period (October to March) and distribution area (northern tier states) is proposed to export 'Malama' avocados from Hawai'i to the continental United States.

Molecular and behavioral studies reveal differences in olfaction between winter and summer morphs of Drosophila suzukii.

Spotted-wing drosophila, Drosophila suzukii (Matsumura), is a major economic pest of several fruit crops in Europe, North and South America, and other parts of the world because it oviposits in ripening thin-skinned fruits. This vinegar fly exhibits two distinct morphotypes: a summer and a winter morph. Although adaptations associated with the winter morph enhance this invasive pest's capacity to survive in cold climates, winter is still a natural population bottleneck. Since monitoring early spring populations is important for accurate population forecasts, understanding the winter morph's response to olfactory cues may improve current D. suzukii management programs. In this study, a comparative transcriptome analysis was conducted to assess gene expression differences between the female heads of the two D. suzukii morphs, which showed significant differences in 738 genes (p ≤ 0.0001). Out of twelve genes related to olfaction determined to be differentially expressed in the transcriptome, i.e., those related to location of food sources, chemosensory abilities, and mating behavior, nine genes were upregulated in the winter morph while three were downregulated. Three candidate olfactory-related genes that were most upregulated or downregulated in the winter morph were further validated using RT-qPCR. In addition, behavioral assays were performed at a range of temperatures to confirm a differing behavioral response of the two morphs to food odors. Our behavioral assays showed that, although winter morphs were more active at lower temperatures, the summer morphs were generally more attracted to food odors. This study provides new insights into the molecular and behavioral differences in response to olfactory cues between the two D. suzukii morphs that will assist in formulating more effective monitoring and physiological-based control tools.

Cold acclimation increases Asian citrus psyllid Diaphorina citri (Hemiptera: Liviidae) survival during exposure to freezing temperatures.

The Asian citrus psyllid, Diaphorina citri, is the vector of the pathogenic bacteria that causes Huanglongbing, the most devasting disease affecting citrus worldwide. As cultivation of citrus tends to expand northward, the tolerance of psyllids to freezing temperatures needs to be investigated. While mortality of D. citri to cold temperature has been previously studied, much less is known regarding the acclimation potential of psyllids to cold temperatures. We first evaluated cold resistance of D. citri depending on color morph and sex. Subsequently, we compared mortality of D. citri at -4 °C for 10 hours between unacclimated psyllids, psyllids that were gradually acclimated to cold over 1 or 2 weeks, and psyllids intermittently acclimated also for 1 or 2 weeks. We did not find major differences in cold resistance between color morphs and sex. We found that after 1 week of gradual acclimation, the survival of psyllids at -4 °C increased to 86%, compared to 20% in the control group. Survival did not increase after the second week of gradual acclimation. Although intermittent acclimation improved survival compared to the control group, it was less effective than gradual acclimation with a survival at 30% and 70% after 1 and 2 weeks of acclimation, respectively, although this difference with gradual acclimation was not significant at week 2. These data show that gradual cold acclimation allows D. citri to survive brief periods below freezing. It may serve as a mechanism responsible for increasing northern establishment of this pest.

Population genomics of Drosophila suzukii reveal longitudinal population structure and signals of migrations in and out of the continental United States.

Drosophila suzukii, or spotted-wing drosophila, is now an established pest in many parts of the world, causing significant damage to numerous fruit crop industries. Native to East Asia, D. suzukii infestations started in the United States a decade ago, occupying a wide range of climates. To better understand invasion ecology of this pest, knowledge of past migration events, population structure, and genetic diversity is needed. In this study, we sequenced whole genomes of 237 individual flies collected across the continental United States, as well as several sites in Europe, Brazil, and Asia, to identify and analyze hundreds of thousands of genetic markers. We observed strong population structure between Western and Eastern US populations, but no evidence of any population structure between different latitudes within the continental United States, suggesting that there are no broad-scale adaptations occurring in response to differences in winter climates. We detect admixture from Hawaii to the Western United States and from the Eastern United States to Europe, in agreement with previously identified introduction routes inferred from microsatellite analysis. We also detect potential signals of admixture from the Western United States back to Asia, which could have important implications for shipping and quarantine policies for exported agriculture. We anticipate this large genomic dataset will spur future research into the genomic adaptations underlying D. suzukii pest activity and development of novel control methods for this agricultural pest.

Does Habituation Affect the Efficacy of Semiochemical Oviposition Repellents Developed Against Drosophila suzukii?

The application of feeding and oviposition repellents is limited to arthropod systems in which habituation does not occur. Although several compounds appear to reduce Drosophila suzukii Matsumura (Dipetra: Drosophilidae) oviposition in berries, previous studies have yet to address whether habituation is a significant risk following preexposure. We tested the response of adult female D. suzukii to three previously identified semiochemical oviposition repellents, 1-octen-3-ol (octenol), ±-geosmin, and 2-n-pentylfuran, following adult and larval preexposure. Using a two-choice gated trap capture assay, we assessed captures in repellent-treated versus blank traps, female survival, and oviposition frequency in the selected trap. We did not find evidence of habituation to octenol or 2-pentylfuran in adult flies preexposed for 24, 48, or 72 hr. When exposed to each of the repellents as larvae, D. suzukii showed similar deterrence as those exposed as adults alone. However, mortality did decrease in F1 octenol treated flies. In contrast with previous investigations we did not observe repellent effects in response to geosmin. Our results suggest that neither exposure during the adult life stage nor during larval development inhibited the effectiveness of octenol and 2-pentylfuran. However, greater survivorship on octenol treated baits in F1 flies, combined with apparent neurotoxic effects of this compound, indicate that octenol may be less suited for field applications. For this reason, 2-pentylfuran appears to be a better candidate for ongoing research aimed at developing an effective push-pull system of behavioral management.

Drosophila suzukii (Diptera: Drosophilidae): A Decade of Research Towards a Sustainable Integrated Pest Management Program.

Drosophila suzukii (Matsumura) (Diptera: Drosophilidae) also known as spotted-wing drosophila (SWD), is a pest native to Southeast Asia. In the last few decades, the pest has expanded its range to affect all major European and American fruit production regions. SWD is a highly adaptive insect that is able to disperse, survive, and flourish under a range of environmental conditions. Infestation by SWD generates both direct and indirect economic impacts through yield losses, shorter shelf life of infested fruit, and increased production costs. Fresh markets, frozen berries, and fruit export programs have been impacted by the pest due to zero tolerance for fruit infestation. As SWD control programs rely heavily on insecticides, exceedance of maximum residue levels (MRLs) has also resulted in crop rejections. The economic impact of SWD has been particularly severe for organic operations, mainly due to the limited availability of effective insecticides. Integrated pest management (IPM) of SWD could significantly reduce chemical inputs but would require substantial changes to horticultural management practices. This review evaluates the most promising methods studied as part of an IPM strategy against SWD across the world. For each of the considered techniques, the effectiveness, impact, sustainability, and stage of development are discussed.

Winter warm-up frequency and the degree of temperature fluctuations affect survival outcomes of spotted-wing drosophila winter morphotypes.

Among overwintering Drosophila suzukii, discrete environmental changes in temperature and photoperiod induce a suite of biochemical changes conferring cold tolerance. However, little is known regarding how temperature fluctuations, which can influence metabolic and cellular repair activity, affect survival outcomes in this species. For that reason, we designed three experiments to test the effects of intermittent warm-up periods and the degree of temperature fluctuation on winter-morphotype (WM) D. suzukii survival. We found that at 5 °C, a temperature sufficient to induce reproductive diapause, but warm enough to allow foraging, increasing warm-up frequency (warmed to 25 °C at various interval schedules) was associated with decreased survival. In contrast, when the nightly low temperature was 0 °C, daily fluctuations that warmed the environment to temperatures above freezing (5, or 15 °C) appeared beneficial and resulted in improved survival compared to flies held at 0 °C during day and night. When we next evaluated cold tolerance using a 24-hour stress test assay (-5 °C), we found that again, thermal fluctuations improved survival compared to static freezing conditions. However, we also found that WM D. suzukii exposed to freezing temperatures during acclimation were less cold tolerant, regardless of the thermal fluctuation schedule, indicating that there may be tradeoffs between adequate acclimation temperature, which is required to induce cold tolerance, and the ensuing effects of incidental chill injury. Moving forward, these data, which account for the nuanced interactions between the thermal environment and in the internal physiology of D. suzukii, may help refine seasonal populations models, which aim to forecast pest pressure based on conditions the previous winter.

Automated aerosol puffers effectively deliver 1-OCTEN-3-OL, an oviposition antagonist useful against spotted-wing drosophila.

BACKGROUND: Automated aerosol puffers releasing behaviorally active volatile organic compounds can deter insect pests in crops. During 2016, we tested the efficacy of aerosol puffer arrays emitting 1-octen-3-ol at reducing Drosophila suzukii oviposition in fall-bearing raspberries in Western New York State. During 2017, we compared the performance of aerosol puffers with a passive diffusion release method (vial dispensers), as well as puffer timing and placement within the field. RESULTS: During 2016, we found that octenol application in the field via aerosol puffer systems resulted in a 20% decrease in D. suzukii oviposition compared to control plots. During 2017, we found that aerosol puffers releasing octenol were 42-55% more effective than vial dispensers at deterring oviposition. We also found that a discontinuous application of octenol during dawn and dusk was sufficient to deter D. suzukii oviposition equivalent to continuous applications throughout the day. Although we observed no differences in infestation depending on puffer placement, low fly populations at the time of the trial may have affected the data. CONCLUSIONS: Our data indicate that automated aerosol puffer systems may reduce D. suzukii infestation to a greater extent than vial dispensers. If adopted, a discontinuous puffer release schedule may protect both economic and ecological interests by reducing the amount of material required to achieve efficacy. Further research on puffer placement is needed to determine whether perimeter applications are effective in larger scale field research and in combination with attractants as part of a push-pull management system.

2-Pentylfuran: a novel repellent of Drosophila suzukii.

BACKGROUND: Drosophila suzukii (Matsumura), spotted-wing drosophila (SWD), is a major invasive pest of soft-skinned fruits in North America and Europe. Although insecticides are currently the primary method of SWD control, it is imperative to develop alternative management approaches, such as behavioral control through the use of repellents and attractants. This study explores the repellent properties of 2-pentylfuran as an oviposition deterrent on raspberries. RESULTS: 2-Pentylfuran was found to be aversive to SWD in laboratory multiple-choice tests. When co-released from a vial (loaded as neat compound) with a synthetic SWD lure, 2-pentylfuran reduced SWD attraction to the SWD lure by 98% and the effect appeared 17% stronger compared to 1-octen-3-ol, a known SWD repellent. Releasing 50% 2-pentylfuran mixed with mineral oil from a vial located near ripe raspberries resulted in 30% reduction in SWD oviposition in the field. In laboratory no-choice assays, 2-pentylfuran reduced SWD oviposition on raspberries above 2.5 mg h-1 with greater repellency achieved at higher release rates. A release rate of 10 mg h-1 from a polyethylene sachet reduced egg-laying on raspberries by 60% in a semifield cage choice experiment. In a field experiment using fruiting raspberry clusters, 14 mg h-1 release rate of 2-pentylfuran was effective at reducing SWD infestations by 56% compared to untreated plots. CONCLUSION: 2-Pentylfuran acts as a repellent for SWD and can significantly reduce fruit infestations under field conditions and high SWD pressure. Given that 2-pentylfuran is a registered food additive and generally regarded as safe, 2-pentylfuran has a potential use in behavioral control strategies against SWD. Published 2020. This article is a U.S. Government work and is in the public domain in the USA.

Seasonal polyphenism of spotted-wing Drosophila is affected by variation in local abiotic conditions within its invaded range, likely influencing survival and regional population dynamics.

Overwintering Drosophila often display adaptive phenotypic differences beneficial for survival at low temperatures. However, it is unclear which morphological traits are the best estimators of abiotic conditions, how those traits are correlated with functional outcomes in cold tolerance, and whether there are regional differences in trait expression.We used a combination of controlled laboratory assays, and collaborative field collections of invasive Drosophila suzukii in different areas of the United States, to study the factors affecting phenotype variability of this temperate fruit pest now found globally.Laboratory studies demonstrated that winter morph (WM) trait expression is continuous within the developmental temperature niche of this species (10-25°C) and that wing length and abdominal melanization are the best predictors of the larval abiotic environment.However, the duration and timing of cold exposure also produced significant variation in development time, morphology, and survival at cold temperatures. During a stress test assay conducted at -5°C, although cold tolerance was greater among WM flies, long-term exposure to cold temperatures as adults significantly improved summer morph (SM) survival, indicating that these traits are not controlled by a single mechanism.Among wild D. suzukii populations, we found that regional variation in abiotic conditions differentially affects the expression of morphological traits, although further research is needed to determine whether these differences are genetic or environmental in origin and whether thermal susceptibility thresholds differ among populations within its invaded range.

Interactions Between Biotic and Abiotic Factors Affect Survival in Overwintering Drosophila suzukii (Diptera: Drosophilidae).

Drosophila suzukii Matsumura is an invasive species affecting berry crops and cherries throughout North America, South America, and Europe. Previous research suggests that in temperate climates, the overwintering success of D. suzukii is likely dependent on access to food, shelter, and adequate cold hardening. We performed a multi-state study under field conditions for two winters to determine whether D. suzukii sex, phenotype (summer-morphotype, winter-morphotype), and life stage (adults, pupae) affected survival over time while recording naturally-occurring spatial and temporal variation in temperature. Access to food was provided and the flies were buried under leaf litter. Baited traps were deployed to determine whether local populations of D. suzukii were active throughout the winter season. The duration of exposure, mean daily temperature, and cumulative time below freezing significantly affected survival. Below freezing, D. suzukii survival was significantly reduced, particularly in northern locations. In contrast, we observed sustained survival up to 10 wk in southern locations among adults and pupae. Biotic factors also significantly affected survival outcomes: female survival was greater than male survival, winter-morphotype survival was greater than summer-morphotype survival, and adult survival was greater than pupal survival. In the north, wild D. suzukii were captured only in early winter, while in the south they were found throughout the winter. These data suggest that although adult D. suzukii may overwinter in sheltered microclimates, this ability may be limited in regions where the ground temperature, or site of overwintering, falls below freezing for extended durations.

Phenotypic Plasticity Promotes Overwintering Survival in A Globally Invasive Crop Pest, Drosophila suzukii.

Spotted wing drosophila, Drosophila suzukii Matsumura, is a major pest of small fruit worldwide in temperate and subtropical growing regions. In Northern climates, D. suzukii likely overwinters locally under leaf litter and snow pack, but our understanding of the factors affecting thermal susceptibility is limited. While previous investigations of thermal susceptibility in this species have employed conventional static acclimation protocols, we aimed to determine whether gradual cooling, or dynamic acclimation, may extend the limits of known thermal tolerance by more closely approximating naturally occurring shifts in temperature. First, we assessed survival among adult and pupal D. suzukii using static acclimation. Then, we re-assessed survival using a novel dynamic acclimation method. We found that while static acclimation was sufficient to induce cold tolerance, dynamic acclimation significantly improved survival at temperatures as low as -7.5 °C. Following static acclimation, the lower lethal limit of adult D. suzukii was -1.1 °C in winter morphotype (WM) adults compared to 1.7 °C in non-acclimated summer morphotype (SM) adults. Dynamic acclimation reduced the lower limit to -5 °C in SM flies. At the end of our study 50% of WM flies survived 72 h at -7.5 °C. Below 0 °C pupal survival declined significantly regardless of acclimation procedure. However, pupal acclimation improved survival outcomes significantly compared to non-acclimated pupae, suggesting that while juvenile diapause is unlikely, cold hardening likely benefits those flies which may develop into the overwintering WM population. These data suggest that the degree of cold hardening is proportional to the thermal environment, a finding previously unrecognized in this species. Given the economic impact of this pest, these data may have important implications for offseason population monitoring and management. We discuss how phenotypic plasticity may drive geographical range expansion, and the impact of climate change on the spread of this species.

Male Psyllids Differentially Learn in the Context of Copulation.

In the Asian citrus psyllid, Diaphorina citri Kuwayama, stimulatory cuticular hydrocarbons act as sex pheromone attractants. Male psyllids locate aggregations of females using those olfactory cues, as well as vibrational communication on the plant surface. Although previous research has indicated that learning plays a role in modulating female reproductive behaviors in psyllids, it is unknown whether males similarly use learning to increase the likelihood of copulatory success. We used an olfactometer-based bio-assay to study the effects of experience on male response to female odor. First, we compared male attraction to female odor in virgin and previously mated males. Second, we tested the effect of several modes of experience with a novel odor, vanillin, to determine whether mating, feeding, or general environmental exposure elicited a learned response. We found that male attraction to female odor significantly increased after mating experience. In addition, we found that males learn about odor specifically in the context of mating, rather than feeding or general exposure. Electrophysiological measurements of antennal response to odorants confirmed that mating status did not affect the sensitivity of the peripheral nervous system to volatile stimuli implicating learning at the level of the central nervous system. These results suggest that male response to female odor is not an entirely innate behavior. Males may require mating experience with female conspecifics to develop attraction to those olfactory cues produced by the female and in association with the female's habitat. This adaptive plasticity may allow males to detect females in an ever-changing environment and promote diversification and further specialization on different host genotypes.

The Influence of Learning on Host Plant Preference in a Significant Phytopathogen Vector, Diaphorina citri.

Although specialist herbivorous insects are guided by innate responses to host plant cues, host plant preference may be influenced by experience and is not dictated by instinct alone. The effect of learning on host plant preference was examined in the Asian citrus psyllid, Diaphorina citri; vector of the causal agent of citrus greening disease or huanglongbing. We investigated: a) whether development on specific host plant species influenced host plant preference in mature D. citri; and b) the extent of associative learning in D. citri in the form of simple and compound conditioning. Learning was measured by cue selection in a 2-choice behavioral assay and compared to naïve controls. Our results showed that learned responses in D. citri are complex and diverse. The developmental host plant species influenced adult host plant preference, with female psyllids preferring the species on which they were reared. However, such preferences were subject to change with the introduction of an alternative host plant within 24-48 hrs, indicating a large degree of experience-dependent response plasticity. Additionally, learning occurred for multiple sensory modalities where novel olfactory and visual environmental cues were associated with the host plant. However, males and females displayed differing discriminatory abilities. In compound conditioning tasks, males exhibited recognition of a compound stimulus alone while females were capable of learning the individual components. These findings suggest D. citri are dynamic animals that demonstrate host plant preference based on developmental and adult experience and can learn to recognize olfactory and visual host plant stimuli in ways that may be sex specific. These experience-based associations are likely used by adults to locate and select suitable host plants for feeding and reproduction and may suggest the need for more tailored lures and traps, which reflect region-specific cultivars or predominate Rutaceae in the area being monitored.

Innate and Conditioned Responses to Chemosensory and Visual Cues in Asian Citrus Psyllid, Diaphorina citri (Hemiptera: Liviidae), Vector of Huanglongbing Pathogens.

Asian citrus psyllid (Diaphorina citri) transmits Huanglongbing, a devastating disease that threatens citrus trees worldwide. A better understanding of the psyllid's host-plant selection process may lead to the development of more efficient means of monitoring it and predicting its movements. Since behavioral adaptations, such as associative learning, may facilitate recognition of suitable host-plants, we examined whether adult D. citri could be conditioned to visual and chemosensory stimuli from host and non-host-plant sources. Response was measured as the frequency of salivary sheaths, the residue of psyllid probing activity, in a line of emulsified wax on the surface of a test arena. The psyllids displayed both appetitive and aversive conditioning to two different chemosensory stimuli. They could also be conditioned to recognize a blue-colored probing substrate and their response to neutral visual cues was enhanced by chemosensory stimuli. Conditioned psyllids were sensitive to the proportion of chemosensory components present in binary mixtures. Naïve psyllids displayed strong to moderate innate biases to several of the test compounds. While innate responses are probably the psyllid's primary behavioral mechanism for selecting host-plants, conditioning may enhance its ability to select host-plants during seasonal transitions and dispersal.