Mitochondrial genome datasets for the sweetpotato weevil, Cylas formicarius elegantulus (Coleoptera: Brentidae), collected in the United States.

The sweetpotato weevil, Cylas formicarius elegantulus (Summers) (Coleoptera: Brentidae), is one of the most destructive pests of sweetpotato worldwide. Genomic analyses of sweetpotato weevils can provide insights into their genetic diversity, population structure, and dispersal as well as provide information to support management strategies. Adult sweetpotato weevils were collected by various methods from Ipomoea batatas L. (sweetpotato) or I. coccinea L. (red morning glory) in the U.S. states of Georgia, Hawaii, South Carolina, and Texas. Genomic DNA was extracted from individual weevil specimens and sequenced using Illumina NovaSeq. A total of 181 GB of 150 base pair (bp) paired-end reads were generated for 40 specimens. Mitochondrial genomes were assembled for each specimen via reference mapping and annotated using Geneious Prime. Full mitochondrial genome sequences range from 17,141 to 17,152 bp with an average GC content of 21.8% and average coverage of 3307 × . A maximum likelihood phylogenetic analysis considering the mitochondrial protein coding genes is provided. Mitochondrial genomes and assembled reads are deposited in NCBI GenBank, providing 40 mitogenomes of C. formicarius elegantulus collected in the U.S.

Optimization of 13-tetradecenyl acetate sex pheromone for trapping Melanotus communis (Coleoptera: Elateridae).

Corn wireworm, Melanotus communis Gyllenhal (Coleoptera: Elateridae), is an economically important larval pest of root and tuber crops in the United States. Previous work to estimate field-level abundance of M. communis has focused on grain-based larval baits placed in soil. However, this sampling method is labor intensive and may not estimate population size accurately. Recent discovery of the M. communis sex pheromone, 13-tetradecenyl acetate, provides a new method to monitor this pest during the adult stage. Early studies with this pheromone showed that different trapping methods might enhance catch and improve trap servicing. We hypothesized that placing lures on elevated traps would increase M. communis capture relative to the in-ground pitfall trapping that is currently used. We had 2 objectives for this study: (a) to compare pheromone captures among in-ground pitfall traps, on-ground pitfalls, elevated pitfalls (1 m), or elevated sticky cards (1 m) and (b) test lure longevity by aging the lures outdoors at 8-, 6-, 4-, 2-, and 0-wk intervals prior to trap deployment in the field. Experiments were conducted in North Carolina, Virginia, South Carolina, and Florida during the 2021 and 2022 field seasons. Results highlight large variation in M. communis abundance across the 4 states. We showed that 1 m elevated pheromone traps caught the most beetles. The age of the lure prior to deployment had a significant effect on trap catch. The lures that were aged for fewer weeks attracted significantly more beetles, with 0- and 2-wk-old lures capturing the greatest numbers.

Buckwheat Flower Volatiles Attract Peristenus spretus and Enhance Its Field-Level Parasitism of Apolygus lucorum.

Volatile compounds play indispensable roles in the interactions among host plants, herbivores and natural enemies. Previous studies showed that the addition of buckwheat strips in cotton fields could attract Peristenus spretus, the dominant parasitoid of Apolygus lucorum, and enhance its parasitic activity. Through the combined analysis of Y-tube olfactometer, solid-phase microextraction (SPME), gas chromatography-mass spectrometer (GC-MS) and electroantennography (EAG), we found that male and female P. spretus responded to compounds present in buckwheat flowers. The five major components of buckwheat flowers, cis-3-hexenyl acetate (Z3HA), 4-methylanisole, 4-oxoisophorone, p-methylphenol and 2-ethylhexyl salicylate, all had a significant attraction to P. spretus adults and led to positive electroantennogram responses, especially for 10 mg/mL 4-oxoisophorone, indicating the components played a key role in the selection behavior of P. spretus to buckwheat flowers. Additionally, field trials showed that the five volatiles could significantly increase the parasitism by P. spretus. Our study screened the key active components of buckwheat flower volatiles that have an attractive effect on P. spretus, revealing its behavioral selection mechanism and emphasizing the important role of plant volatiles on host selection and parasitism of parasitic wasps, providing a theoretical basis for the development of attractants for P. spretus and the reduction of pesticides in the field to promote conservation biological control (CBC) of A. lucorum.

Effect of Host Size on Susceptibility of Melanotus Communis (Coleoptera: Elateridae) Wireworms to Entomopathogens.

Wireworms, the soil-borne larvae of click beetles (Coleoptera: Elateridae), are important crop pests throughout the world. In the eastern U.S., Melanotus communis larvae attack grain, root/ tuber, and vegetable crops. Our objectives were to characterize the pathogenicity and virulence of fungal and nematode entomopathogens on M. communis wireworms, and determine if wireworm size affected virulence. Pathogens tested included five entomopathogenic nematodes, Steinernema carpocapsae (All strain), S. feltiae (SN strain), S. riobrave (355 strain), Heterorhabditis bacteriophora (VS strain), and H. indica (HiHom1 strain); and two entomopathogenic fungi, Beauveria bassiana (GHA strain) and Cordyceps javanica (WF-GA17 strain). None of the pathogens tested caused >15% mortality at 7 or 14 days post-inoculation. Mortality was highest in S. carpocapsae (All strain); the other entomopathogens did not cause higher mortality than the untreated control. Overall, smaller wireworms were more susceptible than larger wireworms. Our results suggested that M. communis wireworms have defenses that limit the ability of the entomopathogens we tested to infect the wireworms. Conceivably, other entomopathogen strains or species may be more effective. Natural populations of entomopathogens may contribute to wireworm population reduction, but further studies are warranted before entomopathogens can be used for M. communis management.

A Symmetrical Diester as the Sex Attractant Pheromone of the North American Click Beetle Parallelostethus attenuatus (Say) (Coleoptera: Elateridae).

Hexanoic acid, 1-octanol, 1,8-octanediol, octyl hexanoate, 1,8-octanediol monohexanoate, and 1,8-octanediol dihexanoate were identified in headspace volatiles collected from the crushed abdomen of a female click beetle of the species Parallelostethus attenuatus (Say) (Elaterinae, tribe Elaterini). In field trials carried out in Illinois, South Carolina, North Carolina, and Virginia, adult male beetles were strongly attracted to 1,8-octanediol dihexanoate alone. Blends of the dihexanoate with one or more of the other compounds proved to be less attractive than the dihexanoate alone, suggesting that the pheromone of this species may consist of a single compound. The symmetrical diester structure of the pheromone is a novel natural product and appears to be structurally unique among insect pheromones.

Volatiles from Cotton Plants Infested by Agrotis segetum (Lep.: Noctuidae) Attract the Larval Parasitoid Microplitis mediator (Hym.: Braconidae).

Herbivore-induced plant volatiles (HIPVs), chemicals produced by plants infested by herbivorous insects, can act as kairomones that recruit natural enemies of the pest herbivore. Agrotis segetum (Denis and Schiffermüller) is a common, important pest of seedling cotton in Xinjiang Province, China, and the braconid Microplitis mediator (Haliday) is an important mortality factor of this pest's larvae. In olfactometer tests, which included healthy foliage, infested foliage, or infested roots, M. mediator preferred A. segetum-infested cotton plants to healthy cotton plants. In GC-MS analyses of plant-emitted volatiles, we found that compounds emitted increased 14.9- and 13.3- fold after leaf infestation and root infestation, respectively, compared to healthy control plants. The volatiles were mainly p-xylene, nonanal, tetradecane, decanal, benzaldehyde, ß-caryophyllene, and humulene, while linalool was only present in the leaf-infestation treatment. In addition, principal component analysis indicated that all 18 compounds were associated with the infested plants, especially ß-caryophyllene, p-xylene, and decanal. Based on the above studies and previous functional evaluations of the volatile compounds, it can be demonstrated that these compounds play a crucial role in modulating the interactions between A. segetum and M. mediator and regulating parasitoid behavior. It may be possible to enhance the biological control of A. segetum by M. mediator through the application of HIPVs.

Evaluation of 13-Tetradecenyl Acetate Pheromone for Melanotus communis (Coleoptera: Elateridae) Detection in North Carolina Row Crop Agroecosystems.

Melanotus communis Gyllenhal (Coleoptera: Elateridae) larvae are a common soil-dwelling pest of many crops, including sweet potato, grains, and tobacco. Although many studies have focused on the larval stage of this pest, the seasonal activity and ecology of the adults (click beetles) are not well understood. The overarching goal of this study was to relate M. communis adult activity to host crops in the North Carolina row-crop agroecosystem. To do this, we conducted a two-year study documenting male M. communis activity, using a recently identified sex attractant pheromone, 13-tetradecenyl acetate. This project was divided into two parts: 1) a pheromone assessment study testing the efficacy and specificity of 13-tetradecenyl acetate, and two analogs, 13-tetradecenyl butyrate and 13-tetradecenyl hexanoate, and 2) a landscape survey using traps baited with 13-tetradecenyl acetate. Results of the efficacy study showed that 13-tetradecenyl acetate was the most effective M. communis lure when compared to non-baited control traps or traps baited with the two homologs. The landscape study documented a strong association between M. communis catch and the adjacent crop type. We found that adult M. communis abundance was greatest near corn, followed by sweet potato, and then cotton. Analysis of activity over time found that the peak activity occurred during July. Overall, this project demonstrates the usefulness of pheromone-baited traps in providing new information about M. communis activity.

Large-Scale Seedling Grow-Out Experiments Do Not Support Seed Transmission of Sweet Potato Leaf Curl Virus in Sweet Potato.

Sweet potato leaf curl virus (SPLCV) threatens global sweet potato production. SPLCV is transmitted by Bemisia tabaci or via infected vegetative planting materials; however, SPLCV was suggested to be seed transmissible, which is a characteristic that is disputed for geminiviruses. The objective of this study was to revisit the validity of seed transmission of SPLCV in sweet potato. Using large-scale grow-out of sweet potato seedlings from SPLCV-contaminated seeds over 4 consecutive years, approximately 23,034 sweet potato seedlings of 118 genotype entries were evaluated. All seedlings germinating in a greenhouse under insect-proof conditions or in a growth chamber were free of SPLCV; however, a few seedlings grown in an open bench greenhouse lacking insect exclusion tested positive for SPLCV. Inspection of these seedlings revealed that B. tabaci had infiltrated the greenhouse. Therefore, transmission experiments were conducted using B. tabaci MEAM1, demonstrating successful vector transmission of SPLCV to sweet potato. Additionally, tests on contaminated seed coats and germinating cotyledons demonstrated that SPLCV contaminated a high percentage of seed coats collected from infected maternal plants, but SPLCV was never detected in emerging cotyledons. Based on the results of grow-out experiments, seed coat and cotyledon tests, and vector transmission experiments, we conclude that SPLCV is not seed transmitted in sweet potato.

Plant Volatiles Modulate Seasonal Dynamics between Hosts of the Polyphagous Mirid Bug Apolygus lucorum.

Plant-derived volatiles play a significant role in host selection of phytophagous insects, but their role in seasonal host shifts remain unclear. The polyphagous mirid bug Apolygus lucorum displays marked seasonal host alternation. During summer, volatiles from flowering plants play a key role in A. lucorum foraging. Though A. lucorum adults deposit overwintering eggs on jujube (Ziziphus jujuba) and grape (Vitis vinifera) during autumn, it is unclear whether plant volatiles equally mediate this host selection behavior. During 2015 and 2016, we found that population densities of A. lucorum adults on cotton (Gossypium hirsutum) during August were higher than those in September, whereas the opposite pattern was observed on fruit trees (i.e., jujube and grape). The dispersal factor of the adult population that dispersed from cotton fields during September was higher than in August, whereas opposite patterns were observed in the neighboring jujube/grape orchard. In Y-tube olfactometer trials, A. lucorum adults preferred cotton plant volatiles over fruit tree odors in August, whereas the opposite patterns were found in September. Three electro-physiologically active volatiles (butyl acrylate, butyl propionate and butyl butyrate) were identified from jujube and grape plants. During September, active volatiles are emitted in considerably greater amounts by jujube and grape than in August, while the amount of volatile emissions in cotton decreases in September. Temporal shifts in plant volatile emission thus may modulate host plant foraging of A. lucorum, and appear to guide its colonization of different host plants. Our findings help understand the role of plant volatiles in the host plant selection and seasonal dynamics of polyphagous herbivores.

13-Tetradecenyl acetate, a female-produced sex pheromone component of the economically important click beetle Melanotus communis (Gyllenhal) (Coleoptera: Elateridae).

Species-specific behavior-modifying chemicals have been used for more than 50 years for monitoring and management of insect pests of agriculture and human health. Elaterid beetle larvae are among insect pests in soil that are increasingly problematic, in part due to the lack of effective management strategies. However, little is known about the insect-produced chemicals that mediate the reproductive behavior of these pests. We used chemical and behavioral studies to identify, synthesize, and field test the sex attractant pheromone of adults of Melanotus communis, commonly called the corn wireworm, the larvae of which are economically important pests of U.S. crops. Our results indicated that a single female-produced chemical, 13-tetradecenyl acetate, was strongly attractive to conspecific male beetles, and did not appear to attract other species. In field evaluations, male M. communis exhibited a dose-dependent response to this compound. In a trial comparing different slow-release dispensers, a small rubber septum impregnated with the chemical was as effective as and easier to use than a plastic bag dispenser. Given that the sex attractant of this insect consists of a single compound that can be readily synthesized, its development for monitoring and management of the corn wireworm may be economically feasible.

Perception of and Behavioral Responses to Host Plant Volatiles for Three Adelphocoris Species.

In China, the genus Adelphocoris (Hemiptera: Miridae) includes three dominant pest species (A. suturalis, A. lineolatus and A. fasciaticollis), which cause great damage to cotton, alfalfa and other crops. In this study, we examined the role of the major volatile organic compounds from plants in host location by these three insects. Gas chromatography-electroantennography and gas chromatography/mass spectrometry analyses identified seven electroantennogram (EAG)-active compounds from 11 host plants. Although the insects responded to all of these compounds in EAG trials, some compounds did not elicit behavioral responses in Y-tube olfactometer bioassays. Adelphocoris suturalis adults showed behavioral responses to four EAG-active compounds, n-butyl ether, butyl acrylate, butyl propionate and butyl butyrate. These four compounds, in addition to p-xylene, were also attractive to A. lineolatus adults. However, A. fasciaticollis adults were attracted only by butyl acrylate, butyl propionate and butyl butyrate. In field trials, A. suturalis and A. fasciaticollis were each attracted to five individual compounds (m-xylene, n-butyl ether, butyl acrylate, butyl butyrate and butyl propionate for A. suturalis and m-xylene, butyl acrylate, butyl butyrate, butyl propionate, and 1,8-cineole for A. fasciaticollis). By contrast, A. lineolatus aduts were attracted to six individual compounds, m-xylene, p-xylene, n-butyl ether, butyl acrylate, butyl butyrate, and butyl propionate. These compounds may be important in host plant location by the Adelphocoris species, and may be useful for developing attractants for adults of these species.

Method for DNA Isolation From Sweetpotato Weevil (Coleoptera: Curculionidae) Collected in Pheromone-Baited Traps.

This study provides a protocol for the isolation of high-quality DNA from sweetpotato weevils (Cylas formicarius elegantulus (Summers)) collected from pheromone-baited aerial funnel traps. This study was based on our discovery that a 2-wk collection interval of sweetpotato weevils from pheromone traps did not permit isolation of intact high-quality genomic DNA. To test the effect of collection methods, i.e., sample collection interval and preservation method, on quality of isolated DNA, we placed freshly killed male sweetpotato weevils into aerial funnel traps in the field and removed subsamples at several times thereafter. DNA yield from freshly isolated (day = 0) samples was significantly greater than samples preserved in 70% ethanol or at -20°C, whereas there was no difference between 70% ethanol and -20°C storage. Likewise, DNA yield from freshly isolated (day = 0) samples was significantly greater than for later sampling times. Quality assessment of genomic DNA through gel electrophoresis and polymerase chain reaction (PCR) indicated isolation of high molecular weight DNA for all samples collected at t ≤ 7 d, but that DNA quality was degraded by 14 d. Our goal was to develop a reliable method for isolation of genomic DNA from field-collected sweetpotato weevil suitable for direct use in PCR. We discovered that it is critical to collect specimens from traps at an interval of 1 wk or less. Our findings allow for scheduling of sampling at reasonable intervals without the need for special materials. This has the added benefit of allowing individuals without special training to collect and prepare sweetpotato weevil specimens for genetic studies.

Methyl jasmonate induction of cotton: a field test of the 'attract and reward' strategy of conservation biological control.

Natural or synthetic elicitors can affect plant physiology by stimulating direct and indirect defence responses to herbivores. For example, increased production of plant secondary metabolites, a direct response, can negatively affect herbivore survival, development and fecundity. Indirect responses include increased emission of plant volatiles that influence herbivore and natural enemy behaviour, and production of extrafloral nectar that serves as a food source for natural enemies after their arrival on induced plants. Therefore, the use of elicitors has potential for the study of basic aspects of tritrophic interactions, as well as application in biorational pest control, i.e. an 'attract and reward' strategy. We conducted a field study to investigate the effects of methyl jasmonate, an elicitor of plant defence responses, on three trophic levels: the plant, herbivores and natural enemies. We made exogenous applications of methyl jasmonate to transgenic cotton and measured volatile emission, extrafloral nectar production and plant performance (yield). We also assessed insect abundance, insect performance, and parasitism and predation of brown stink bug, Euschistus servus, eggs in methyl jasmonate-treated and untreated control plots. Application of methyl jasmonate increased emission of volatiles, in particular, (+)-limonene and (3E)-4,8-dimethyl-1,3,7-nonatriene, and production of extrafloral nectar, but not yield, compared with the control treatment. Despite increased volatile and extrafloral nectar production, methyl jasmonate application did not affect plant bug performance, or mortality of E. servus egg masses, and only marginally influenced insect abundance. Mortality of E. servus eggs varied over the course of the study. Overall, methyl jasmonate treatment affected cotton plant-induced responses, but not the insects that inhabit the plants. Our results were probably influenced by reduced natural enemy colonization of cotton from adjacent non-crop habitats, and subsequent low within-field population recruitment. Much remains to be learned about the effects of exogenous application of plant-produced 'enhancers' on the behaviour of natural enemies before crop physiology can be manipulated to enhance pest control.

Volatile fragrances associated with flowers mediate host plant alternation of a polyphagous mirid bug.

Apolygus lucorum (Hemiptera: Miridae) is an important insect pest of cotton and fruit trees in China. The adults prefer host plants at the flowering stage, and their populations track flowering plants both spatially and temporally. In this study, we examine whether flower preference of its adults is mediated by plant volatiles, and which volatile compositions play an important role in attracting them. In olfactometer tests with 18 key host species, the adults preferred flowering plants over non-flowering plants of each species. Coupled gas chromatography-electroantennography revealed the presence of seven electrophysiologically active compounds from flowering plants. Although the adults responded to all seven synthetic plant volatiles in electroantennography tests, only four (m-xylene, butyl acrylate, butyl propionate and butyl butyrate) elicited positive behavioral responses in Y-tube olfactometer bioassays. The adults were strongly attracted to these four active volatiles in multi-year laboratory and field trials. Our results suggest that these four fragrant volatiles, which are emitted in greater amounts once plants begin to flower, mediate A. lucorum's preference to flowering host plants. We proved that the use of commonly occurring plant volatiles to recognize a large range of plant species can facilitate host selection and preference of polyphagous insect herbivore.

Electrophysiological and behavioral characterization of bioactive compounds of the Thymus vulgaris, Cymbopogon winterianus, Cuminum cyminum and Cinnamomum zeylanicum essential oils against Anopheles gambiae and prospects for their use as bednet treatments.

BACKGROUND: Laboratory and field studies showed that repellent, irritant and toxic actions of common public health insecticides reduce human-vector contact and thereby interrupt disease transmission. One of the more effective strategies to reduce disease risk involves the use of long-lasting treated bednets. However, development of insecticide resistance in mosquito populations makes it imperative to find alternatives to these insecticides. Our previous study identified four essential oils as alternatives to pyrethroids: Thymus vulgaris, Cymbopogon winterianus, Cuminum cyminum, Cinnamomum zeylanicum. The objectives of this study were to identify active compounds of these essential oils, to characterize their biological activity, and to examine their potential as a treatment for bednets. METHODS: We evaluated the electrophysiological, behavioural (repellency, irritancy) and toxic effects of the major compounds of these oils against Anopheles gambiae strain 'Kisumu'. RESULTS: Aldehydes elicited the strongest responses and monoterpenes the weakest responses in electroantennogram (EAG) trials. However, EAG responses did not correlate consistently with results of behavioral assays. In behavioral and toxicity studies, several of the single compounds did exhibit repellency, irritancy or toxicity in An. gambiae; however, the activity of essential oils did not always correlate with activity expected from the major components. On the contrary, the biological activity of essential oils appeared complex, suggesting interactions between individual compounds and the insect under study. Data also indicated that the three effects appeared independent, suggesting that repellency mechanism(s) may differ from mechanisms of irritancy and toxicity. CONCLUSIONS: Based on the bioassays reported here, some of the compounds merit consideration as alternative bednet treatments.

Sugar concentration and timing of feeding affect feeding characteristics and survival of a parasitic wasp.

The availability of food sources is important for parasitoid survival, especially for those that inhabit ecosystems where nectar and honeydew are spatially or temporally scarce. Therefore, the value of even a single meal can be crucial for survival. Psyttalia lounsburyi is a parasitoid, and biological control agent, of the olive fruit fly, Bactrocera oleae. In order to improve our understanding of the basic nutritional ecology of P. lounsburyi and its role in survival we evaluated the effect of a single sucrose meal on the longevity of female and male wasps. We measured the duration of feeding, volume ingested, sucrose consumption, energy content, and longevity of wasps provided with different concentrations of sucrose (0.5, 1, and 2M) at different times after emergence (0, 1, 2 or 3 days after emergence). Our results showed that longevity was significantly influenced by sucrose concentration and timing of feeding. For females, feeding on sucrose increased the likelihood of survival to varying degrees, ranging from 32.3% to 95.4%, compared to water-only controls. The longest duration of feeding was observed for the highest sucrose concentrations and oldest wasps. The amount of sugar ingested and energy uptake increased, up to a point, as sugar concentration increased. Our results suggest that P. lounsburyi derived greatest benefit from the intermediate concentration (1M) of sucrose provided 2 or 3 days after emergence. Our study emphasizes the importance of finding balance between increasing longevity and limiting the duration of feeding, and concomitant uptake of nutrients, that is fundamental for survival of the wasp in nature.

Plant volatiles influence electrophysiological and behavioral responses of Lygus hesperus.

Previous laboratory studies have shown that the mirid Lygus hesperus is attracted to volatiles emitted from alfalfa; feeding damage increases the amounts of several of these volatiles, and visual cues can enhance attraction further. The present study tested single plant volatiles in electrophysiological and behavioral trials with L. hesperus. Electroantennogram (EAG) analyses indicated that antennae responded to most plant volatiles included in the test, and that when gender differences were observed, males usually were more responsive than females. Antennal responses to the alcohols ((E)-3-hexenol, (Z)-3-hexenol, 1-hexanol), the acetate (E)-2-hexenyl acetate, and the aldehyde (E)-2-hexenal were among the strongest. Moderate responses were observed for (E)-beta-ocimene, (E,E)-alpha-farnesene, (+/-)-linalool, and methyl salicylate. A dose dependent response was not observed for several terpenes (beta-myrcene, beta-caryophyllene, (+)-limonene, or both (R)-(+)- and (S)-(-)-alpha-pinenes). EAG responses, however, were not always consistent with behavioral assays. In Y-tube bioassays, males did not exhibit a positive behavioral response to any of the compounds tested. Instead, males were repelled by (E)-2-hexenyl acetate, (+/-)-linalool, (E,E)-alpha-farnesene, and methyl salicylate. In contrast, female L. hesperus moved upwind towards (R)-(+)-alpha-pinene, (E)-beta-ocimene, and (E,E)-alpha-farnesene, and showed a negative response towards (Z)-3-hexen-1-ol, (S)-(-)-alpha-pinene, and methyl salicylate. This study emphasizes the use of multiple approaches to better understand host plant finding in the generalist herbivore L. hesperus.

EAG-active herbivore-induced plant volatiles modify behavioral responses and host attack by an egg parasitoid.

Volatiles emitted by plants in response to feeding by Lygus species were tested in neurophysiological, behavioral, and parasitism trials with Anaphes iole, an egg parasitoid of Lygus. Electroantennogram analyses indicated that A. iole antennae responded to most herbivore-induced plant volatiles (HIPVs) tested and that females were usually more responsive than males. Antennal responses to (Z)-3-hexenyl acetate and methyl salicylate were among the strongest. Behavioral assays in a four-arm olfactometer demonstrated that response of female wasps to (Z)-3-hexenyl acetate varied greatly depending on preconditioning regime. Preconditioning wasps to complex host-plant odors led to stronger preference than did a single preconditioning stimulus, i.e., (Z)-3-hexenyl acetate. In a horizontal wind tunnel, female wasps were attracted by methyl salicylate and alpha-farnesene. Parasitism of Lygus lineolaris eggs by A. iole in a cotton field was greater when the eggs were associated with (Z)-3-hexenyl acetate or alpha-farnesene than with controls. Overall, the results of this study show that A. iole can perceive a variety of plant volatiles released after its host damages plants, that the degree of associative learning in A. iole can be manipulated based on preconditioning regime, and that single synthetic HIPVs are attractive to A. iole and can be used to increase attack rates on host eggs. Therefore, it appears that HIPVs have potential for use in suppression of Lygus population densities.

Nutritional ecology of a parasitic wasp: food source affects gustatory response, metabolic utilization, and survivorship.

The success of biological control is partly mediated by the longevity and reproductive success of beneficial insects. Availability of nectar and honeydew can improve the nutrition of parasitic insects, and thereby increase their longevity and realized fecundity. The egg parasitoid, Anaphes iole, showed strong gustatory perception of trehalulose, a carbohydrate found in homopteran honeydew. Chromatographic analysis demonstrated that enzymatic hydrolysis of sucrose, a common nectar sugar, proceeded at a faster rate than that of melezitose, a sugar common in aphid honeydew. A long-term bioassay showed that longevity was greater at 20 degrees C than at 27 degrees C, and at both temperatures survival was generally greatest for wasps provisioned with the three major nectar sugars, sucrose, glucose, and fructose. Patterns of food acceptance and utilization showed that A. iole accepted and utilized a broad range of sugars found in nature, including those found in nectar as well as honeydew. Glucose, fructose, and several oligosaccharides composed of these monosaccharide units appear to be more suitable for A. iole than other sugars tested. Evidence suggests that individual fitness benefits afforded by food sources are important for a time-limited parasitoid, and that continued investigations on the interface between nutrition and biological control are warranted for A. iole.

Incorporation of rhodamine B into male tobacco budworm moths Heliothis virescens to use as a marker for mating studies.

Rhodamine B, a dye commonly used in a variety of biological studies was incorporated into the bodies of male tobacco budworm moths, Heliothis virescens (Lepidoptera: Noctuidae), by allowing them to feed freely on 0.1% rhodamine dissolved in a 10% sucrose solution. After exposing males for one to three days to this pigment, rhodamine was clearly detectable in >82% of spermatophores extracted from untreated females. The intake of this dye did not affect the life span, the production of eggs or the capacity of moths to copulate when compared with moths fed only a sucrose solution or water. Rhodamine B was easily identifiable externally but was more apparent internally in males after only one day of exposure to the pigment. Even at this short feeding duration, rhodamine was detectable in >50% of males 5 days after feeding stopped. Longer exposure to the dye significantly increased the percentage stained. Detection of rhodamine was slightly enhanced by the use of ultraviolet light. The dye accumulation in internal abdominal organs was a better indicator of the presence of the pigment than external contamination of the moth. The use of the method described in this report can be a tool for the rapid incorporation of a low cost dye in the tobacco budworm for biological, behavioral and genetic studies.