This study explored the flower visiting behaviors and pollination abilities of mason bees (Osmia excavata Alfken (Hymenoptera: Megachilidae)), bumble bees (Bombus terrestris (Linnaeus, 1758) (Hymenoptera: Apidae)), and Italian honey bees (Apis mellifera ligustica Spinola (Hymenoptera: Apidae)) in apple orchards in early spring in Jinan (located in the central region of Shandong) and Yantai (located in the Peninsula of Shandong). We compared the pollen collection patterns, flower visiting behavior, flying speed, and effects on apple pollination of the 3 types of bees. The frequencies of flower visits were significantly higher for mason bees (12.89/min in Jinan and 10.63/min in Yantai) than bumble bees and Italian honey bees in the 2 regions. The single flower residence times were significantly higher for Italian honey bees (8.22 s in Jinan and 9.43 s in Yantai), but Italian honey bees were most affected by the climate. The 3 bees differed significantly in terms of the amount of apple pollen collected and their effects on the fruit setting rate in apples (mason bees > bumble bees > Italian honey bees). The results showed that the mason bee was the most suitable pollinating species for spring apple orchards; Bumble bees were more suitable as alternative pollinators during cloudy and low temperatures; Italian honey bees were able to take advantage of their large number of worker bees in sunny and warm weather. Compared to individual bee species, a combination of 2 or 3 species of bees might be more advantageous in dealing with complex and variable weather conditions.

Cacopsylla pyricola (Förster) (Hemiptera: Psyllidae) is the most expensive and challenging insect pest of commercial pear trees in the Pacific Northwest. Integrated pest management (IPM) programs are working toward relying more heavily on natural enemies to reduce insecticide use. Trechnites insidiosus (Crawford) (Hymenoptera: Encyrtidae) is the main parasitoid of C. pyricola, but little is known about its biology in the region. Developing sampling tools is important for the deployment of IPM programs, including monitoring of natural enemies. In this study, we examined 2 conventional monitoring methods: beat trays and yellow sticky cards, in addition to screened sticky cards and 3D-printed cylinder traps. Additionally, we tested an overwintering trap for the collection of parasitized C. pyricola.The trapping methods were tested in orchards in Oregon and Washington. Unscreened cards caught the most T. insidiosus and C. pyricola, followed by screened cards, cylinder traps, and then beat trays. Beat trays sometimes failed to catch any T. insidiosus, even when it was found in abundance via other methods. Screened cards and cylinder traps reduced bycatch and increased ease of identifying T. insidiosus. Specimens from the cylinder traps were also more suitable for use in molecular analysis. The overwintering traps were effective at capturing parasitized C. pyricola, but were highly variable year to year. The ideal trapping method will vary based on research needs (e.g., DNA preservation, reducing bycatch, catching higher numbers), but both screened sticky cards and cylinder traps were viable methods for monitoring T. insidiosus and its host.

Agrilus mali stands as a significant wood-boring pest prevalent in Northeast Asia. Identifying this pest beetle is often hindered by insufficient efficient, rapid, on-site discrimination methods beyond examining adult morphological features. As a result, an urgent need arises for developing and implementing a rapid and accurate molecular technique to distinguish and manage the beetle. This study presents a straightforward, swift, highly specific, and sensitive method built upon recombinase polymerase amplification combined with a lateral flow dipstick (RPA-LFD). This method demonstrates the capability to promptly identify the beetle, even during its larval stage. RPA primers and probes were designed using the internal transcribed spacer 1 region. Through probe optimization, false-positive signals were successfully eliminated, with an accompanying discussion on the underlying causes of such signals. The RPA-LFD assays exhibited remarkable specificity and sensitivity, requiring as little as 10^–3 ng of purified DNA. Furthermore, the extraction of crude DNA was achieved through immersion in sterile distilled water, thus streamlining the assay process. Achievable at temperatures ranging from 30 to 50 °C, the RPA-LFD assay can be executed manually without specialized equipment. By merging the RPA-LFD assay with DNA coarse extraction, A. mali can be detected within just 30 min. This current study effectively demonstrates the immense potential of RPA-LFD in quarantine and pest management. Additionally, it presents a universal technique for the rapid on-site diagnosis of insects, showcasing the wide applicability of this method.

Graphical Abstract

Ceratitis is an economically important genus of fruit flies that originated in Africa, has a wide host range, and causes serious economic losses due to its invasive damage. As a result, it is critical to identify them accurately and quickly in the world. Loop-mediated isothermal amplification (LAMP), as one of the representatives of isothermal amplification technology, has been widely used in the rapid nucleic acid detection of human pathogens and has shown its advantages in the identification of insect agricultural pests. In this study, using the mitochondrial cox1 and cob genes as target genes, the rapid molecular identification of the Ceratitis FARQ complex, C. cosyra, and C. capitata was realized based on LAMP. The experimental conditions optimization results showed that F3/B3:FIP/BIP = 1:8 was the optimal primer concentration ratio and 63 °C was the optimal reaction temperature. The sensitivity of the primers obtained in this study can reach up to 0.01 ng/µl DNA. A loop-mediated isothermal amplification identification technology system was established based on rapid, rough DNA extraction and visual detection of Ceratitis economically important fruit flies.The positive reaction system changed from pink to khaki by visual detection.The identification flow can be completed within 1 hour, including sample processing, DNA extraction, and LAMP visual detection.

Trading partners often require phytosanitary or quarantine treatments for fresh horticultural produce to ensure no economically important pest species are moved with the imported product. When developing such treatments, it is essential that the level of treatment efficacy can be determined.This is often based on the mortality of the total number of target pests exposed to treatment, but in naturally infested products this number is not always known. In such cases, the infestation rate and subsequently an estimate of the number of pests are obtained directly from a set of untreated control samples of the host product.The International Plant Protection Convention (IPPC) Secretariat has provided 2 formulas for these situations that place an interval around the point estimate obtained from the control samples to obtain an estimate of the infestation rate. However, these formulas do not allow a confidence level to be assigned to the estimate, and there are concerns with the assumptions regarding the distribution and the measure of variability used in the formulas. In this article, we propose 2 alternative formulas. We propose that the lower one-sided confidence limit should be applied to all infestation datasets that are approximately normally distributed. As infestation data are sometimes skewed, it is proposed the lower one-sided modified Cox confidence limit is applied to data approximately log-normal distributed.These well-recognized formulas are compared to the formulas recommended by the IPPC and applied to 3 datasets involving natural infestation.

Wireworms are primary pests of potatoes in Canada with relatively few effective control options. Recently, a new meta-diamide insecticide, broflanilide, was registered in Canada and the United States as an in-furrow spray applied at planting that provides protection of tubers from feeding damage and dramatically reduces wireworm populations. As part of our routine screening of wireworm response to novel insecticides, we exposed wireworms (predominantly Agriotes obscurus; N=2320) to field soil collected from plots to which either bifenthrin (Capture 2EC) or broflanilide (Cimegra) had been applied at registered rates 124–145 and 314–335 days previously in 2018, 2019, and 2021. Wireworm behavioral responses were assessed using a previously developed numerical scale, and indicated residues were present in sufficient quantity in all broflanilide and most bifenthrin-treated plots to induce morbidity. Transfer of affected wireworms to untreated soil indicated morbidity was generally reversible after exposure to bifenthrin, but not after exposure to broflanilide. There was an inverse relation between wireworm size and the degree of morbidity induced by exposure to broflanilide, but not bifenthrin. Analyses of soil residues indicated readily quantifiable levels of broflanilide still present in undisturbed (not harvested) field soil 314–335 days after application. Insecticide residues in soil samples from disturbed (harvested) sections of the potato plots were lower, as was the degree of morbidity of wireworms exposed to this soil. The use of wireworms as bioindicators of insecticide residues, and the implications of insecticide persistence for wireworm management are discussed.

Soybean gall midge, Resseliella maxima Gagné, was recently identified as a new species causing significant injury to soybean and is currently found in 164 counties across 7 midwestern states (NE, IA, SD, MN, MO, ND, and KS). Infestation of soybean begins in late spring, when adults emerge from last year's soybean field. Infestation of a new soybean crop depends on the presence of fissures which start to form at the base of the soybean plant around the V2 stage. Field observations indicate that these fissures are only present below the cotyledonary nodes or in the area within 3–5 cm above the soil surface.To determine the importance of these fissures for R. maxima infestation and plant injury, hilling or the movement of the soil to cover the base of soybean plants at the V2–V3 stage was compared with the standard practice (no-hilling). Field studies were conducted at 3 sites in east-central Nebraska during the 2021 growing season.The results showed a significant reduction in the frequency of infested plants, larval number per plant, and plant injury for hilled compared to no-hill treatment. This reduction in the presence of larvae and plant injury corresponded with a significantly greater yield for hilled compared to the no-hill treatment. These results highlight the importance of fissures on soybean for R. maxima adult infestation as well as the potential for hilling to be used as a management strategy for R. maxima.

This study investigated the seasonal occurrence of bark and woodboring Coleoptera in Pinus densiflora (Pinales: Pinaceae), and Larix kaempferi (Pinales: Pinaceae) stands using multifunnel traps baited with pine volatiles in Korea. The number and species of bark and woodboring beetles caught in traps baited with ethanol, α-pinene, and ethanol+α-pinene were compared to determine the effective attractants. In addition, the effects of other pine volatiles, such as (-)-β-pinene, β-caryophyllene, (±)-limonene, β-myrcene, and 3-carene, were investigated. A total of 13,134 woodboring beetles from 150 species were collected from pine and larch stands from 2019 to 2020. Tomicus minor (Hartig) (Coleoptera: Curculionidae) adults were more attracted to traps baited with α-pinene, whereas Xyleborinus saxesenii (Ratzeburg) (Coleoptera: Curculionidae), Cyclorhipidion pelliculosum (Eichhoff) (Coleoptera: Curculionidae), and Phloeosinus pulchellus (Blandford) (Coleoptera: Curculionidae) adults were more attracted to traps baited with ethanol. Hylurgops interstitialis (Chapuis) (Coleoptera: Curculionidae), Shirahoshizo genus group, Rhagium inquisitor (Linne) (Coleoptera: Cerambycidae), and Rhadinomerus maebarai (Voss & Chûjô) (Coleoptera: Curculionidae) were more frequently attracted to traps baited with ethanol+α-pinene than to traps baited with other attractants.The addition of 3-carene to ethanol+α-pinene enhanced the capture of H. interstitialis, R. inquisitor, and Hylobius (Callirus) haroldi (Faust) (Coleoptera: Curculionidae).

Reticulitermes speratus (Kolbe) is economically important pest in East Asia including Korea, Japan, and China where they infest wooden structures in urban areas. Previously, it has been reported that R. speratus consists of 5 subspecies, R. speratus kyushuensis Morimoto, R. speratus speratus Kolbe, R. speratus leptolabralis Morimoto, R. speratus okinawanus Morimoto, R. speratus yaeyamanus Morimoto, while only R. speratus kyushuensis was recorded in Korea in the past. However, it remains elusive if different subspecies of R. speratus other than R. speratus kyushuensis are present in Korea. In this study, we report the first record of R. speratus speratus from Korea, which was verified using soldier morphology and molecular characteristics obtained from a mitochondrial gene. R. speratus speratus Kolbe, 1885 (Blattodea: Rhinotermitidae) are found in several provinces, mainly southern regions in Korea, whereas R. speratus kyushuensis are distributed throughout the country. Our morphological comparison showed that R. speratus speratus can be distinguishable from R. speratus kyushuensis by the ratio of the posterior postmentum width to length. In the molecular comparison, R. speratus speratus revealed genetic differences of 3.06% (range 2.60–4.10%) from R. speratus kyushuensis using cytochrome oxidase subunit II gene sequences.

Adult male cerambycid beetles of the subfamilies Cerambycinae and Lamiinae emit aggregation-sex pheromones that attract both sexes, and these chemicals can be utilized for quarantine surveillance for related exotic species which produce the same or similar pheromones. Here, we assess how attraction of 7 cerambycid species to pheromone-baited traps was influenced by the release rates of synthesized pheromones from polyethylene sachet emitters. Compounds tested included racemic 3-hydroxyhexan-2-one, the (R)-enantiomer of which is the sole or major pheromone component of numerous cerambycine species, and 2 compounds that are pheromone components of many lamiine species: (E)-6,10-dimethyl-5,9-undecadien-2-yl acetate (fuscumol acetate) and 6-methylhept-5-en-2-ol (sulcatol). We confirmed release rates of these compounds could be manipulated by varying the doses loaded into emitters. Various doses and concentrations, ranging from very dilute to the neat compounds, were then tested in field trials. Several species were most strongly attracted to the highest dose of a tested compound, one species to an intermediate dose, while another species was equally attracted regardless of dose. Our results demonstrate the importance of characterizing dose–response relationships for cerambycid species targeted by pheromone-based trapping.

The thrips Megalurothrips usitatus Bagnall and Frankliniella intonsaTrybom (Thysanoptera:Thripidae) are important pests in cowpea-growing areas of south China. Parthenogenesis is an important reproductive form of thysanopterans, and plays an important role in maintaining population growth. To understand the developmental and reproductive characteristics of these 2 thrips species during parthenogenesis, we compared the age–stage life tables of M. usitatus and F. intonsa on cowpea pods under natural regimes during the summer and winter.The results showed that the total preadult period and total preoviposition period of M. usitatus were significantly longer than those of F. intonsa in both seasons. Moreover, longevity of adult M. usitatus (29.53 days) was shorter compared with adult F. intonsa (34.00 days) in summer, whereas higher fecundity (220.8 eggs/female) and more oviposition days (37.83 days) were observed in M. usitatus compared with F. intonsa in winter (fecundity = 179.83 eggs/female, oviposition days = 33.03 days).The net and gross reproductive rates of M. usitatus were significantly greater than those of F. intonsa during winter. In addition, the intrinsic and finite rates of increase of M. usitatus were significantly lower than those of F. intonsa, and the mean generation time of M. usitatus was significantly longer than that of F. intonsa both in summer and winter.These results indicated that parthenogenesis has species specificity among thrips, which in turn affects population development, especially under changing environments.

Understanding orchard floor management is critical to organic tree-fruit production systems given its impact on weeds, soil fertility, tree health, and crop yield. Several viable options are available to producers for weed management and promotion of organic fertility, including use of turf and broadleaf alleyway covers and living and nonliving tree-row mulches. While these measures can be effective, little is known about how these strategies influence arthropod pests, which cause fruit injury. Here, we assessed 6 organic orchard floor management strategies for their impact on arthropod abundance and diversity in an organic peach production system in northern Utah from 2010 to 2014, using sweep netting and pitfall collections along with observed peach fruit damage. Generally, we found that alleyway and tree-row treatments had no impact on total arthropod diversity, species richness, or community diversity. However, earwig (Forficula auricularia) abundance was significantly impacted by alleyway and tree-row treatments that resulted in increased fruit injury. Trefoil alleyway treatments consistently increased earwig abundance across life-history stages, while mulch or Alyssum (straw) tree-row treatments harbored more earwigs and, as a result, increased earwig fruit injury. Since earwigs are especially prone to damaging young, developing fruits, it is imperative that more work is done to assess earwig abundances and life-history traits. Our results demonstrate that detrimental arthropods are sensitive to orchard floor management and can further inform integrated pest management approaches that complement sustainability goals.

Lycorma delicatula White, commonly called spotted lanternfly, is an invasive fulgorid that was first detected in the United States in Pennsylvania in 2014 and has spread across Eastern states. Lycorma delicatula is a phloem feeder, and feeding of persistent populations can cause wilt and dieback in host plants. Vineyards in infested regions have experienced vine loss, where damage has been attributed to L. delicatula. Nymphal and adult lifestages have been observed on cultivated apple and peach trees, but the feeding effects of this pest on these crops have not been quantified. Here, we caged young potted apple and peach trees and infested them with 50 nymphal or 25 adult L. delicatula per plant for 3 months and tracked plant health. Horticultural measurements including trunk diameter, height, leaf drop, and photosynthesis rates were recorded before, during, and after the infestation. To evaluate the long-term effects of infestation, all apple and peach trees were removed from pots and planted in an orchard block where their phenology, growth, and physiology could be evaluated the following growing season. Short- and long-term measurements showed no significant differences in apple tree health between infested and noninfested trees. There was a significant reduction in the growth of trunk diameter in peach trees during exposure to early-instar nymphs; infested peach trees also had significantly increased frost damage to buds the following spring. However, there were no long-term impacts on peach growth and physiology. These results indicate that L. delicatula likely will not be a major threat to these deciduous fruit crops.

The hala scale, Thysanococcus pandani Stickney, is an invasive insect that infests hala trees, Pandanus tectorius, and has recently arrived in the Hawaiian Islands of Maui, Oahu, and Molokai. Hala scale is native to Thailand, Indonesia, and Singapore and was accidentally introduced to Maui, with its first detection in Hana in 1995. The hala tree is a frequently planted ornamental plant throughout Hawaii's urban landscape and is also a native tree found in coastal areas of ecological suitability throughout the state. These trees are integral to native Hawaiian weaving traditions, as the leaves were used to make items such as sails, hats, or mats.The spread of T. pandani threatens the continuation of these practices, as the plant material may not be sufficient for weaving. Hala trees infested with T. pandani can have negative effects such as deformation, discoloration, and early tree death. Experimental field trials were set up to evaluate the efficacy of organic and chemical insecticides as potential treatments to mitigate the damage of T. pandani infestations. This was done on the Island of Maui, where T. pandani is widespread. These trees were artificially inoculated with T. pandani and evaluated throughout the trials for changes in plant height, canopy width, scale removal, and visual infestation ratings. The results of 2 repeated field trials indicated that flupyradifurone and buprofezin were effective treatments in the reduction of the hala scale infestation and improving visual aspects related to plant health.

The invasive Halyomorpha halys (Hemiptera: Pentatomidae) has threatened Mid-Atlantic tree fruit since 2010. To identify factors underlying observed differences in H. halys pest pressure among individual orchards within a geographically proximate area, a 3-yr study was conducted across 10 apple orchard and 8 nonorchard sites bordered by unmanaged woodlots. At each site, 3 pheromone traps were monitored weekly for H. halys captures from late April to mid-October. Apple injury was assessed at harvest at orchard sites annually, and a survey of woody plants found in woodlots adjacent to all sites was conducted. There were no significant differences in captures between orchard and nonorchard site types, but captures were significantly different among individual orchard sites and among individual nonorchard sites. A significant positive relationship between the amount of stink bug injury on apple at harvest and late season captures was detected at orchard sites in 2018 and 2019. Among woodlots adjacent to all sites, a significant positive relationship between the proportion of Lonicera spp. and mid- and late-season nymphal captures was identified. Season-long nymphal captures were positively related to the proportion of Lonicera and Elaeagnus and negatively with Sassafras. For adults, captures were negatively related to the proportion of Ailanthus and positively related to the proportion of Fraxinus in the early and mid-season, respectively. Our results indicate that orchard presence was not driving the relative abundance of localized H. halys populations and that differences in relative densities among sites point to other factors, such as abundance of specific uncultivated woody hosts in unmanaged areas.

Knowledge of specialty crop cultivars with resistance against insect pests is limited, and this may serve as a barrier to implementing host-plant resistance as part of an integrated pest management strategy. Carrot (Daucus carota L.) (Apiaels: Apiaceae)is a valuable specialty crop with a diversity of insect pests and cultivars that differ in physical and chemical qualities that influence insect pest preferences. To investigate the role of cultivar as a tool to reduce insect pest damage, we evaluated 7 carrot cultivars in replicated laboratory and field trials in IN and OH, USA in 2021. During June and July, we documented oviposition and feeding damage by the carrot weevil (Listronotus oregonenesis LeConte) (Coleoptera: Curculionidae) and used faunistic analysis to measure the abundance and diversity of foliar insect assemblages on each cultivar. We found no significant differences in oviposition and root damage across cultivars in the field, with mean cumulative egg scars ranging from 1.83 ± 1.40 in “Red Core Chantenay” to 5.17 ± 2.62 in “Cosmic Purple”. However, there was a positive correlation between the cumulative number of egg scars and number of trichomes on petioles. Similarly, no-choice laboratory bioassays revealed no significant differences in mean cumulative egg scars, ranging from 5.00 ± 1.15 in “Red Core Chantenay” to 10.63 ± 1.02 in “Danvers 126”. Predominant insect pests differed across cultivars, but Cicadellidae was common across all cultivars. Interestingly, only 1 beneficial insect family, Pteromalidae, was predominant across cultivars.This research highlights the impact of cultivar selection on the diversity and damage potential of insect pests in carrot production.

The production of almonds and pistachios in California's Central Valley employs insecticides for the management of their primary pest, navel orangeworm.The pyrethroid Bifenthrin is commonly used, and now a strain of Amyelois transitella Walker (Lepidoptera: Pyralidae) (R347) obtained from Kern County almond orchards with a history of Bifenthrin use has acquired >110-fold resistance toward pyrethroids. One method to improve control is to use additives and spray adjuvants, which are applied simultaneously with an insecticide to increase coverage and/or duration of control. We tested 2 levels of the naturally occurring clay Kaolin as an additive, alone and in combination with either Bifenthrin or the diamide Chlorantraniliprole, to determine if it could reduce feeding damage and decrease survival of pyrethroid-resistant A. transitella on almonds in the laboratory and improve the efficacy of Chlorantraniliprole in the field. Larval performance was measured for the strains R347 and ALM (34.7-fold resistance compared to susceptible lab strain) reared on treated almonds. Strain R347 had 1.9-fold greater survival and caused 1.3-fold more feeding damage than strain ALM across all treatments, although both strains were susceptible to the combination of Kaolin + insecticide. Kaolin synergized Bifenthrin for R347, decreasing survival by 10.0%. Kaolin did not reduce feeding damage for either strain. When combined with insecticide, feeding damage was similar to insecticide alone, but the addition of Kaolin to the insecticide generally decreased survival more than the insecticide alone. In the field, the addition of Kaolin to Chlorantraniprole during application helped retain activity against this challenging pest.

Exclusion netting in some European and North American apple (Malus domestica Borkhausen, Rosaceae, Rosales) orchards has been documented to be an effective method of control for multiple insect pest species. By minimizing reliance on insecticides, these orchards have reduced costs, risks to the environment and nontarget species, and reduced the risk of insecticide resistance. This study examined the use of commercially available hail netting (DrapeNet®; Prosser, WA) as a pest exclusion strategy under conditions in Minnesota, USA. In 2021 and 2022, we assessed the efficacy of hail netting as a tool for pest suppression in orchards by monitoring pest species in netted and open plots crossed with and without insecticide applications. Our findings show that both of the major pest species in Minnesota, the codling moth (Cydia pomonella L.; Lepidoptera:Tortricidae) and the apple maggot (Rhagoletis pomonellaWalsh; Diptera:Tephritidae), were significantly reduced inside the netting compared to open plots by 94% and 96%, respectively. For a secondary pest, the red-banded leafroller (Argyrotaenia velutinana Walker; Lepidoptera: Tortricidae), moth populations were reduced by 56%. We also found that insecticide application alone did not significantly reduce pest pressure in these species. Additionally, we investigated the subsequent effects of hail netting on fruit quality and yield. The use of hail netting and insecticide application resulted in significantly higher proportions of high-quality fruit at harvest. However, netting did not significantly influence yield.These findings suggest that hail netting can be used to control Midwest apple insect pests with limited insecticide applications while maintaining high fruit quality.

Onion thrips (Thrips tabaci Lindeman,Thysanoptera:Thripidae) is a significant insect pest of onions (Allium cepa L., Asparagales: Amaryllidaceae). In addition to feeding on onion foliage, they may spread plant pathogens. Currently, onion thrips and pathogens are managed as separate pests with insecticides and fungicides. It may be beneficial to manage these pests simultaneously as limiting onion thrips may reduce pathogen damage. We tested combinations of bio- and conventional pesticides in a season-long management program in Michigan onion fields. From 2020 to 2022, we counted onion thrips weekly and visually estimated plant foliage necrotic damage (%) in experimental plots each year. In 2020, we tested 6 treatment programs including: azadirachtin, spinosad, a copper-based fungicide, azadirachtin + copper-based fungicide, spinosad + copper-based fungicide, and untreated control. The thrips populations were not significantly reduced compared to the control, but necrotic damage was reduced significantly in spinosad-treated plots. In 2021, we tested a combination of 8 bio- and conventional pesticide programs. Compared to the control, the bioinsecticides did not reduce onion thrips populations, but the conventional pesticide programs reduced both onion thrips numbers and necrotic damage. In 2022, we tested only conventional insecticide programs but included 3 different action thresholds for initiation and applied them with or without a fungicide, for 8 treatments. All insecticide programs reduced onion thrips compared to the control, the action threshold did not impact thrips numbers significantly. Overall, the use of action thresholds can lead to fewer insecticide applications and a lower incidence of leaf damage.

Severe bermudagrass mite (Aceria cynodoniensis Sayed) infestation stunts turfgrass growth and reduces the aesthetic and recreational value of managed bermudagrass. Management practices, such as fertilization, mowing, and irrigation, may impact bermudagrass mite infestation and damage, but empirical evidence is lacking. Two 20 wk experiments were conducted with potted bermudagrass in a greenhouse or nursery to evaluate the effect of varying nitrogen rates (0, 24.5, or 49 kg N/ha), mowing heights (1.3, 2.5, 3.8, or 5 cm), and irrigation rates (60%, 100%, or 140% evapotranspiration [ET] rate) on the densities of witch's brooms (i.e., stunted and deformed terminals symptomatic of infestation) and bermudagrass mites. Increasing nitrogen fertility from 0 to 49 kg N/ha increased witch's broom and bermudagrass mite densities by 292% and 339%, respectively. Bermudagrass fertilized with nitrogen maintained higher turf quality than unfertilized grass despite greater mite damage. Decreasing irrigation from 140% to 60% of the ET rate also increased witch's broom densities by 124%. Mowing height did not consistently affect witch's broom or mite densities. Witch's broom and mite densities were positively correlated and followed a general trend with greater densities in April–August and a decline in densities in August–October. These findings suggest that nitrogen fertilization and water stress influence bermudagrass mite damage.Thus, limiting nitrogen fertilization to a level necessary to maintain turfgrass health and quality (0.5 kg N/ha) and minimizing turfgrass water stress can complement current chemical control strategies as part of an integrated pest management program.

Subterranean termites in the genus Reticulitermes Holmgren 1913 are among the most economically important wood-destroying pests in the western United States. Yet, there remains uncertainty regarding the taxonomy and biology of the species in this genus. The 2 species described as having distributions in this region are the western subterranean termite, Reticulitermes hesperus Banks, and the arid land subterranean termite, Reticulitermes tibialis Banks. Taxonomic studies utilizing cuticular hydrocarbon (CHC) profiles, agonistic behavior, flight phenology, and mitochondrial DNA (mtDNA) suggested that R. hesperus is a species complex comprised of 2 or more sympatric, yet reproductively isolated species. To further delineate these taxa, we examined multiple genes from samples of Reticulitermes collected in the western United States. Alates collected after recent spring and fall mating flights, as well as previously collected workers, were subjected to CHC phenotyping and DNA sequence analyses that targeted mitochondrial cytochrome oxidase subunit II (COII), mitochondrial 16S rRNA, and nuclear Internal Transcribed Spacer 1 and 2 (ITS1 and 2). Phylogenetic analyses conducted also included published sequences of other putative western Reticulitermes species. Results suggest that at least 5 species of Reticulitermes may be present in California and that Reticulitermes in Arizona consistently group into multiple clades, including samples previously identified as R. tibialis in a sister clade. These analyses further support the species status of qualitatively different CHC phenotypes and that alates swarming in spring vs. fall are reproductively isolated species.

Lepidopteran pests have been successfully managed by the adoption of insect resistant transgenic plants expressing Cry and/or Vip insecticidal proteins derived from Bacillus thuringiensis (Bt plants). Among such pests, Spodoptera frugiperda (Smith, 1797) (Lepidoptera: Noctuidae) is highlighted for its destructive potential in maize crops and for cases of field-evolved resistance to Bt plants. Cry insecticidal proteins expressed in Bt plants are known for their interaction with insect midgut receptors and subsequent midgut cell disruption that leads to target pest death. In the midgut of lepidopteran larval pests such as S. frugiperda, serine proteases are important in dietary protein digestion and activation or degradation of insecticidal proteins. This work was conducted to evaluate if the use of a soybean trypsin inhibitor (SBTI) could disrupt the development of a Bt-susceptible and a Bt-resistant population of S. frugiperda ingesting Bt (expressing Cry1F, Cry1A.105, and Cry2Ab2 Cry proteins) and non-Bt maize plants. The SBTI was produced and purified using recombinant expression in E. coli followed by purification in Ni-Sepharose. Bioassays using non-Bt maize leaves indicated that the development of susceptible and resistant populations of S. frugiperda was not influenced by the ingestion of SBTI. However, when the resistant population consumed Bt maize plants amended with SBTI, high mortality along with a reduction in larval weight and reduced activity of digestive trypsins were observed. Although the mode of action was not elucidated, it is possible that the consumption of SBTI increased susceptibility to Bt maize in the resistant population of S. frugiperda.

Annual indoxacarb resistance in Helicoverpa armigera (Hübner) populations collected from various crops in Australia was monitored between 2013 and 2023. Resistance frequency determined by F₂ screening using a predetermined discriminating dose of indoxacarb, was lowest in the 2013–2014 and 2015–2016 seasons at 0.0164 and 0.0246, respectively. Resistance then increased significantly to a ten-year high of 0.0869 in 2018–2019 but declined to 0.0557 in 2019–2020 during a severe drought, remaining relatively stable thereafter to 2023. Indoxacarb resistance was first detected in H. armigera collected from maize in the Gwydir valley, New South Wales, in 2013 (strain GY7-39). In 2017, a second indoxacarb resistant H. armigera strain (UN1U3-10) was isolated from a population collected in chickpeas in the Liverpool Plains, New South Wales. Indoxacarb resistance of this strain was characterized to evaluate its potential to compromise the ongoing effectiveness of insecticide resistance management strategies in Australian farming systems. Survival at the discriminating dose of indoxacarb in UN1U3-10 was 28.9, 52.6, 86.7, and 92.9% in the F₂, F₃, F₄, and F₅, respectively. Following introgression with a susceptible strain and reselection with the discriminating dose of indoxacarb, the resistance ratio of UN1U3-10 was approximately 800-fold. Resistance was autosomal, incompletely dominant and conferred by more than 1 locus. While indoxacarb resistance in UN1U3-10 did not confer to emamectin benzoate or spinetoram and there was no evidence of major cross-resistance to the Bt toxins Cry1A, Cry2A or Vip3A, there was 5-fold reduced sensitivity to chlorantraniliprole. Indoxacarb resistance was suppressed by approximately 10-fold by PBO with no synergism byTPP or DEM, suggesting the involvement of cytochrome P450 enzymes. A stability analysis indicated a fitness cost may be associated with the genes that confer resistance in the UN1U3-10 strain.The potential risk for diverse indoxacarb resistance in the Australian H. armigera population is discussed.

The Gill's mealybug, Ferrisia gilli Gullan, (Hemiptera: Pseudococcidae) has emerged as a major pest of pistachio in California. Because F. gilli is only relatively recently described, there are no validated reference genes to normalize the expression data from real-time quantitative reverse transcription–polymerase chain reaction (qRT-PCR) in this species. We selected and validated 8 commonly used reference genes (RPS8, TBP, UBQE2, RPL7, RPL5, RPL40, RPLP1, and HEL) for expression stability in F. gilli. These genes were evaluated in 5 different geographical populations of F. gilli collected from organic and conventionally grown pistachio orchards. Candidate reference genes were also evaluated in F. gilli fed with 4 plant hosts: pistachio, almond, grapes, and lima beans.The stability of candidate genes was analyzed using 4 software algorithms: geNorm, NormFinder, BestKeeper, and RefFinder.Three genes RPS8, RPL40, and RPL7encoding for ribosomal proteins were identified as the most stable across the treatments and thus were recommended for normalizing the qRT-PCR data.These findings will support resistance monitoring, molecular toxicology, and functional genomics research in F. gilli.

Piezodorus guildinii (Westwood, 1837) (Hemiptera: Pentatomidae) is an important arthropod pest of soybean (Glycine max (L.) Merr.) throughout American continents. However, the historical events associated with its dispersion are poorly understood. In this study, we employed a phylogeographic approach to investigate the origin and demographic history of P. guildinii in Brazil. We analyzed the cytochrome c oxidase subunit I and Cytb gene sequences of P. guildinii individuals collected in Brazil's 5 soybean production macro-regions and cross-referenced this information with sequences available in public databases. Our findings support an older Caribbean basin establishment for the current genealogical strains of P. guildinii, with subsequent dispersion to Brazil around 0.97 Mya. No secondary dispersion of this species from the Caribbean region to soybean areas in Brazil was identified. The Brazilian populations of P. guildinii are genetically structured across the country's soybean macro-regions and show strong signals of continuous demographic and spatial expansion in Brazil, which may be accelerated by the soybean cropping landscape in the country.The populations from the northern region (MR5) are older than the Central and South populations.The signs of demographic expansion indicate that P. guildinii populations are increasing their effective size in soybean regions, which could reflect its importance as a soybean pest in the coming years.

Graphical Abstract

Western corn rootworm, Diabrotica virgifera virgifera (LeConte) (Coleoptera: Chrysomelidae), is the most serious economic pest of maize, Zea mays (L.) (Poales: Poaceae), in the U.S. Corn Belt and also threatens production in Europe. Traditional management options have repeatedly failed over time as western corn rootworm rapidly develops resistance to insecticides, transgenic maize and even crop rotation. Traits that improve host plant resistance and tolerance are highly sought after by plant breeders for crop protection and pest management. However, maize resistance to western corn rootworm appears to be highly complex and despite over 75 yr of breeding efforts, there are no naturally resistant hybrids available commercially. Using phenotypic data from field and greenhouse experiments on a highly diverse collection of 282 inbred lines, we screened and genetically mapped western corn rootworm-related traits to identify genetic loci which may be useful for future breeding or genetic engineering efforts. Our results confirmed that western corn rootworm resistance is complex with relatively low heritability due in part to strong genotype by environment impacts and the inherent difficulties of phenotyping below ground root traits. The results of the Genome Wide Associated Study identified 29 loci that are potentially associated with resistance to western corn rootworm. Of these loci, 16 overlap with those found in previous transcription or mapping studies indicating a higher likelihood they are truly involved in maize western corn rootworm resistance. Taken together with previous studies, these results indicate that breeding for natural western corn rootworm resistance will likely require the stacking of multiple small effect loci.

The potential for population genomics to elucidate invasion pathways of a species is limited by taxonomic identification issues. The Oriental fruit fly pest, Bactrocera dorsalis (Hendel) belongs to a complex in which several sympatric species are attracted to the same lure used in trapping and are morphologically cryptic and/or reported to hybridize. In this study, we evaluated the taxonomic ambiguity between B. dorsalis and 2 major cryptic species, based on morphological expertise and 289 target specimens sampled across the whole distribution range. Specimens were then subjected to DNA sequence analyses of the COI mitochondrial barcode and the EIF3L nuclear marker to evaluate the potential for molecular identification, in particular for specimens for which morphological identification was inconclusive. To this aim, we produced reference datasets with DNA sequences from target specimens whose morphological identification was unambiguous, which we complemented with 56 new DNA sequences from closest relatives and 76 published and curated DNA sequences of different species in the complex. After the necessary morphological observation, about 3.5% of the target dataset and 47.6% of the specimens from Southeast Asian islands displayed ambiguous character states shared with B. carambolae and/or B. occipitalis. Critical interpretation of DNA sequence data solved morphological ambiguities only when combining both mitochondrial and nuclear markers. COI discriminated B. dorsalis from 5 species; EIF3L and ITS from another species. We recommend this procedure to ensure correct identification of B. dorsalis specimens in population genetics studies and surveillance programs.

The leaffooted bug, Leptoglossus clypealis Heidemann, is a seed-feeding economic pest of crops including almonds and pistachios. The historical distribution of L. clypealis has been considered to be West of the Mississippi in the United States. L. clypealis was recently found in sorghum in the Coastal Bend of Texas, representing a new host record and new collection locality. This study investigated the genetic diversity of L. clypealis museum voucher samples from the Western United States (i.e., Texas, California, and Idaho) collected from 1994 to 2019, including the L. clypealis samples from the Coastal Bend. Eleven new sequences were obtained. Sample sequences were compared with public sequences of L. clypealis from the Western United States. The mitochondrial DNA cytochrome oxidase 1 (mtDNA COI) barcode gene region revealed differences among and within the collection regions. Texas, Idaho, and California all had samples with unique genotypes, and the combined dataset had a haplotype diversity of 1.0. The Texas specimens recently collected in the Coastal Bend did not match genotypes from California or Idaho, and it is unlikely they were recently introduced. Overall, L. clypealis from Texas, Idaho, and California have a high level of genetic diversity, and the 3 regions appear to be within the native range of the species.

As Lycorma delicatula (White) continues to spread across the United States, more winegrapes are potentially susceptible to damage from this pest. Lycorma delicatula, spotted lanternfly, is primarily associated with Ailanthus altissima (Mill.) Swingle, a tree from its native range that is now globally distributed. While L. delicatula is a known pest of cultivated Vitis spp. in South Korea, its relationship with the specific grape species grown in the United States is unclear. This study assessed L. delicatula survivorship and development on 5 Vitis species, including 2 winegrape V. vinifera L. varieties, ‘Pinot Noir’ and ‘Chardonnay’, Concord grape, Vitis labrusca L., River grape, Vitis riparia Michx., and muscadine grape, Vitis rotundifolia Michx. var. ‘Carlos’. A diet of A. altissima served as a positive control. Lycorma delicatula provided with a diet of V. riparia or V. vinifera ‘Pinot Noir’ yielded the highest survivorship and fastest rates of development among grape diets and were statistically equivalent to those provided with A. altissima. Vitis rotundifolia did not support L. delicatula growth past the third-instar life stage, indicating this species is a poor host for the early development of this pest. Our results indicate that both V. riparia and V. vinifera are favorable hosts for L. delicatula and may provide the means for this insect to invade and establish in new regions.