Acibenzolar-S-methyl induces resistance against ambrosia beetle attacks in dogwoods exposed to simulated flood stress.

Xylosandrus spp. ambrosia beetles (Coleoptera: Curculionidae: Scolytinae) are important wood-boring pests of nursery trees weakened by abiotic and biotic stressors. Acibenzolar-S-methyl (ASM), a plant defense elicitor, was tested for inhibiting Xylosandrus spp. tunneling (i.e., attacks) into flood-stressed flowering dogwoods (Cornus florida L. (Cornales: Cornaceae)). Container-grown dogwoods were treated with ASM substrate drench + flooding, ASM foliar spray + flooding, ASM drench + no flooding, ASM foliar + no flooding, no ASM + flooding, or no ASM + no flooding at 3 days before flood stress in a completely randomized design under field conditions. Trees were flooded for 14 days and then drained and watered as needed. Attacks were counted every 2 days for 28 days. Plant tissue samples were collected at 7 and 14 days after flooding to determine ethanol content using solid-phase microextraction-gas chromatography-mass spectrometry. Trees were dissected to determine gallery formation and depth, fungal colonization, and the presence of eggs, larvae, and adults. The highest number of Xylosandrus beetle species attacks were recorded from plants exposed to no ASM + flooding, but attacks were reduced in ASM treated trees (drench or foliar) + flooding. Trees treated with drenches had fewer attacks than foliar sprays. Plants assigned to no flood had the fewest beetle attacks. Moreover, ASM reduced Xylosandrus spp. gallery formation and depth, fungal colonization, and presence of eggs, larvae, and adults. All flooded trees produced ethanol. In conclusion, ASM induced a plant defense response to Xylosandrus spp. tunneling in dogwoods under flood stress conditions.

Characterization of Solenopsis invicta virus 4, a polycipivirus infecting the red imported fire ant Solenopsis invicta.

Solenopsis invicta virus 4 (SINV-4), a new polycipivirus, was characterized in the host in which it was discovered, Solenopsis invicta. SINV-4 was detected in the worker and larval stages of S. invicta, but not in pupae, male or female alates, or queens. The SINV-4 titer was highest in worker ants, with a mean of 1.14 × 107 ± 5.84 ×107 SINV-4 genome equivalents/ng RNA. Electron microscopic examination of negatively stained samples from particles purified from SINV-4-infected fire ant workers revealed isometric particles with a mean diameter of 47.3 ± 1.4 nm. The mean inter-colony SINV-4 infection rate among S. invicta worker ants was 45.8 ± 38.6 in Alachua County, Florida. In S. invicta collected in Argentina, SINV-4 was detected in 22% of 54 colonies surveyed from across the Formosa region. There did not appear to be any seasonality associated with the SINV-4 infection rate among S. invicta nests. SINV-4 was successfully transmitted to uninfected S. invicta colonies by feeding. Among three colonies of S. invicta inoculated with SINV-4, two retained the infection for up to 72 days. The replicative genome strand of SINV-4 was detected in 18% (n = 11) of SINV-4-infected S. invicta colonies. Among 33 ant species examined, the plus genome strand of SINV-4 was detected in undetermined species of Dorymyrmex and Pheidole, Cyphomyrmex rimosus, Monomorium pharaonis, Pheidole obscurithorax, Solenopsis geminata, Solenopsis richteri, Solenopsis xyloni, and Solenopsis invicta. However, the replicative (minus) genome strand was only detected in S. invicta. SINV-4 infection did not impact brood production or queen fecundity in S. invicta. The mean brood rating (63.3% ± 8.8) after 31 days for SINV-4-infected colonies was not statistically different from that of uninfected colonies (48.3 ± 25.5). At the end of the 31-day test period, mean egg production was not significantly different between SINV-4-infected S. invicta colonies (287.7 ± 45.2 eggs laid/24 hours) and uninfected control colonies (193.0 ± 43.6 eggs laid/24 hours).

Field evaluation of Solenopsis invicta virus 3 against its host Solenopsis invicta.

Viruses have been used successfully as biocontrol agents against several insect pests but not ants. Laboratory tests have shown that Solenopsis invicta virus 3 (SINV-3) may be an effective natural control agent against its host, the red imported fire ant (Solenopsis invicta Buren). In this field trial, SINV-3 was released into 12 active S. invicta nests within a 0.088-hectare area in Florida and the impact on the ants monitored. SINV-3 was successfully transmitted, established, and multiplied within treated colonies reaching a maximum mean value of 8.71 × 108 ± 8.26 × 108 SINV-3 genome equivalents/worker ant 77 days after inoculation. SINV-3 was not detected in any of the nests in the control group. A 7-fold decrease in nests was observed in the SINV-3-treated group compared with the untreated control. A correspondingly significant decrease in S. invicta nest size also was observed over the course of the evaluation. Based on the nest rating scale, nest size among those treated with SINV-3 decreased from 3.92 ± 1.24 on day 0 to 1.67 ± 2.06 on day 77, which represents a 57.4% decrease in size. Conversely, the nest rating for the control group increased 9.3%, from 4.42 ± 1.24 on day 0 to 4.83 ± 2.12 on day 77. A follow-up survey of SINV-3-treated and -untreated plots conducted 9 months after initial treatment revealed that fire ant populations rebounded, but at a different rate. A total of 11 and 19 nests were detected in the SINV-3-treated and -untreated areas, respectively. SINV-3 was still detected in the treated area 1.8 years after the initial virus treatment and the virus had spread into the adjacent control plot. Results demonstrate that SINV-3 is an effective natural control agent against the invasive ant, S. invicta; the virus causes no known detrimental ecological impacts, is host specific, and sustained in the environment.

Comparative Performance of Fungicides, Biofungicides, and Host Plant Defense Inducers in Suppression of Phytophthora Root Rot in Flowering Dogwood During Simulated Root Flooding Events.

During flooding events in nurseries, Phytophthora root rot caused by Phytophthora cinnamomi Rands often causes damage that leads to complete crop loss. In this study, we evaluated the efficacy of fungicides, biofungicides, and host plant defense inducers for preventive and curative control of Phytophthora root rot on flowering dogwood (Cornus florida L.) seedlings exposed to a simulated flood event of 1, 3, or 7 days. In two greenhouse trials, preventive (7 days before flooding) or curative (1 day after flooding) drench treatments were applied to dogwood seedlings artificially inoculated with P. cinnamomi. The plants were flooded by maintaining standing water for 1, 3, or 7 days. After the trials, plant growth data (total plant weight, root weight, plant height, and plant width) were recorded, and root systems were assessed for disease severity using a scale of 0 to 100% of roots affected, and subsamples were plated on PARPH-V8 medium to determine the percent recovery of the Phytophthora pathogen. Plants preventively treated with Subdue MAXX had reduced disease severity relative to the nontreated, inoculated plants (positive control) flooded 1, 3, or 7 days in both trials. Pageant Intrinsic and Segovis treatments also had lower disease severity than the positive control at all flooding durations in trial two, but not trial one. In trial one, preventive and curative treatments of Orkestra Intrinsic had reduced disease severity compared with the positive control at 1 and 3 days of flooding, whereas curative treatments of Empress Intrinsic and Tartan Stressgard also were effective at 1 and 3 days of flooding in trial one. The host plant defense inducers (Aliette 80 WDG, Signature Xtra, and Actigard) were inconsistent and ineffective at reducing disease severity when applied as preventive or curative treatments. Preventive treatments of the biofungicides RootShield Plus+ and MBI-110 had consistently lower disease severity than the positive control at 1 day of flooding but not 3 or 7 days of flooding. Potentially, growers can use information from this study to manage Phytophthora root rot during flooding or in areas of the nursery that often experience high soil moisture levels.

Movement of Xylosandrus germanus (Coleoptera: Curculionidae) in Ornamental Nurseries and Surrounding Habitats.

Some exotic ambrosia beetles are damaging pests in ornamental nurseries. Xylosandrus germanus (Blandford) is the most problematic ambrosia beetle in Ohio nurseries. Movement of X. germanus in nurseries has not been characterized, and knowledge is lacking on whether infestations originate from within nurseries or surrounding habitats. Flight activity of X. germanus was monitored in nurseries and adjacent wooded areas to determine the source of beetles infesting nurseries, and characterize their movement within nurseries. Ethanol-baited bottle traps were positioned within wooded areas adjacent to commercial nurseries and within nurseries at various distances from the nursery woodlot interface. Flight activity of overwintered X. germanus occurred in wooded areas adjacent to nurseries before occurrence within nurseries. There was a direct relationship between degree-days and the distance from woodlots when X. germanus were first found in traps in spring, with earlier captures closest to wooded areas and latest ones furthest away into the nursery. X. germanus appeared to move into nurseries from adjacent wooded areas, with numbers trapped within nurseries decreasing with distance away from wooded areas. Trees in the interior of nurseries would appear to be subjected to less attack pressure than trees near the nursery border. Intercepting beetles as they move into nurseries might be an effective strategy to reduce attack pressure on valuable trees.

Monitoring attack and flight activity of Xylosandrus spp. (Coleoptera: Curculionidae: Scolytinae): the influence of temperature on activity.

Wood-boring ambrosia beetles (Coleoptera: Curculionidae: Scolytinae), including Xylosandrus spp., are key pests in ornamental nurseries. Knowledge of their activity in spring is important for nursery growers to effectively time their protective sprays. We measured the reliability of ethanol-baited bottle traps for monitoring emergence of overwintered Xylosandrus spp. in ornamental nurseries. Detection of initial flight activity by traps was compared with initial attacks on ethanol-injected trap trees. To develop tools for forecasting Xylosandrus germanus (Blandford) activity, the relationships between temperature and their attack and flight activity were examined, and the bloom sequence of ornamental plants was examined as phenological indicators of X. germanus emergence in Ohio. Captures of X. germanus coincided with attacks on trap trees on seven of eight occasions over 2 yr in four nurseries. Xylosandrus crassiusculus (Motshulsky) were detected in only one nursery and captures coincided with attacks each year. There was a strong relationship between maximum daily temperatures 20 and 21degrees C and X. germanus attack and flight activity. No attack or flight activity were detected in a monitoring period unless there were 1 or 2 d of at least 20 degrees C. Emergence of X. germanus always began after and within 6 d of full bloom on Cornelian cherry dogwood, and usually after and within 4 d of first bloom on Norway maple and full bloom on border forsythia. The traps or phenological indicators can be used by growers to monitor emergence of X. germanus to time their initial protective sprays. The relationship between X. germanus activity and temperature can be used by growers to make decisions on timing subsequent treatments.

Ethanol injection of ornamental trees facilitates testing insecticide efficacy against ambrosia beetles (Coleoptera: Curculionidae: Scolytinae).

Exotic ambrosia beetles are damaging pests in ornamental tree nurseries in North America. The species Xylosandrus crassiusculus (Motshulsky) and Xylosandrus germanus (Blandford) are especially problematic. Management of these pests relies on preventive treatments of insecticides. However, field tests of recommended materials on nursery trees have been limited because of unreliable attacks by ambrosia beetles on experimental trees. Ethanol-injection of trees was used to induce colonization by ambrosia beetles to evaluate insecticides and botanical formulations for preventing attacks by ambrosia beetles. Experiments were conducted in Ohio, Tennessee, and Virginia. Experimental trees injected with ethanol had more attacks by ambrosia beetles than uninjected control trees in all but one experiment. Xylosandrus crassiusculus and X. germanus colonized trees injected with ethanol. In most experiments, attack rates declined 8 d after ethanol-injection. Ethanol-injection induced sufficient pressure from ambrosia beetles to evaluate the efficacy of insecticides for preventing attacks. Trunk sprays of permethrin suppressed cumulative total attacks by ambrosia beetles in most tests. Trunk sprays of the botanical formulations Armorex and Veggie Pharm suppressed cumulative total attacks in Ohio. Armorex, Armorex + Permethrin, and Veggie Pharm + Permethrin suppressed attacks in Tennessee. The bifenthrin product Onyx suppressed establishment of X. germanus in one Ohio experiment, and cumulative total ambrosia beetle attacks in Virginia. Substrate drenches and trunk sprays of neonicotinoids, or trunk sprays of anthranilic diamides or tolfenpyrad were not effective. Ethanol-injection is effective for inducing attacks and ensuring pressure by ambrosia beetles for testing insecticide efficacy on ornamental trees.

Acute toxicity of plant essential oils to scarab larvae (Coleoptera: Scarabaeidae) and their analysis by gas chromatography-mass spectrometry.

Larvae of scarab beetles (Coleoptera: Scarabaeidae) are important contaminant and root-herbivore pests of ornamental crops. To develop alternatives to conventional insecticides, 24 plant-based essential oils were tested for their acute toxicity against third instars of the Japanese beetle Popillia japonica Newman, European chafer Rhizotrogus majalis (Razoumowsky), oriental beetle Anomala orientalis (Waterhouse), and northern masked chafer Cyclocephala borealis Arrow. Diluted solutions were topically applied to the thorax, which allowed for calculating LD50 and LD90 values associated with 1 d after treatment. A wide range in acute toxicity was observed across all four scarab species. Of the 24 oils tested, allyl isothiocyanate, cinnamon leaf, clove, garlic, and red thyme oils exhibited toxicity to all four species. Allyl isothiocyanate was the most toxic oil tested against the European chafer, and among the most toxic against the Japanese beetle, oriental beetle, and northern masked chafer. Red thyme was also comparatively toxic to the Japanese beetle, oriental beetle, European chafer, and northern masked chafer. Interspecific variability in susceptibility to the essential oils was documented, with 12, 11, 8, and 6 of the 24 essential oils being toxic to the oriental beetle, Japanese beetle, European chafer, and northern masked chafer, respectively. Analysis of the active oils by gas chromatography-mass spectrometry revealed a diverse array of compounds, mostly consisting of mono- and sesquiterpenes. These results will aid in identifying active oils and their constituents for optimizing the development of plant essential oil mixtures for use against scarab larvae.

Preharvest quarantine treatments of chlorantraniliprole, clothianidin, and imidacloprid-based insecticides for control of Japanese beetle (Coleoptera: Scarabaeidae) and other scarab larvae in the root zone of field-grown nursery trees.

Japanese beetle, Popillia japonica Newman (Coleoptera: Scarabaeidae), is an important quarantine pest of nurseries. Nursery plant movement from P. japonica-infested regions is regulated by the U.S. Domestic Japanese Beetle Harmonization Plan (DJHP), which classifies states by risk categories. Treatments for category 2 states include preharvest soil surface treatment of nursery plants grown in field soil using Discus SC, Marathon (1G or 60 WP), or Flagship (0.22G or 25 WG). In this study, Discus, Marathon 60 WP, or Flagship 0.22G DJHP standards were compared with labeled rates of non-DJHP-approved insecticides, including neonicotinoids clothianidin (Arena 50WDG), generic imidacloprid (Quali-Pro Imidacloprid 2 F T&O Insecticide, Mallet 2 F T&O Insecticide, and Lada 2 F Insecticide), and imidacloprid + bifenthrin (Allectus SC), as well as the anthranilic diamide, chlorantraniliprole (Acelepryn Insecticide). Arena provided 100% P. japonica control in May, June, and July over four test years, but had one larva recovered during August in two of those 4 yr. Acelepryn did not provide DJHP-acceptable P. japonica control. During July, Allectus provided 100% P. japonica control in three of four test years, but had four larvae in one test year. Other treatments tested only during July, which provided 100% P. japonica control, included Discus (five tests); Marathon, Quali-Pro, and Mallet (two tests); and Lada and Flagship (one test). Generic imidacloprid 2 F formulations were equivalent in P. japonica control to DJHP-approved insecticides. Insecticides generally performed poorly on other scarabs or curculionid larvae. The study supports Arena, Allectus, and generic imidacloprid 2 F products as suitable candidates for the DJHP.

Evaluation of winter cover crop methods for management of flatheaded appletree borer (Coleoptera: Buprestidae).

Winter cover crops grown at the base of red maples (Acer rubrum L.) can reduce flatheaded appletree borer (Chrysobothris femorata Olivier; Coleoptera: Buprestidae; FAB) damage by both physically blocking preferred oviposition sites and altering the environment around them. However, cover crop competition negatively affects tree growth. To investigate long-term cover crop effects, trees grown with cover crop for 2 yr were transitioned to a standard herbicide practice. After 4 yr, trees in the initial 2 yr cover crop plots were 1 yr behind in growth compared to trees grown in bare rows for all 4 yr. Most growth reduction occurred in the first year following transplant. Additional borer losses of 1-2% per year were observed in production years 3 and 4. A second experiment answered 2 questions: Can killing the cover crop once it reaches maximal height protect trees from borers while also reducing competition and thereby increase tree growth? and Do herbicide applications increase borer attacks? In this experiment, red maples were grown with (i) a standard herbicide program, (ii) a mulch mat, (iii) a cover crop that was killed early, or (iv) a cover crop allowed to senesce naturally. Evaluations after 2 yr indicated early kill of the cover crop was not enough to improve tree growth. Further, trees in the early kill cover crop treatment had the most FAB attacks. Cover crops allowed to senesce naturally reduced FAB attacks in both studies; however, more work is needed to minimize disparities in tree growth during the initial year post-transplant and determine the causal relationship between herbicide use and borer attacks.

Integration of Control Strategies to Optimize Management of Ambrosia Beetles (Coleoptera: Curculionidae, Scolytinae) and Phytophthora Root Rot (Peronosporales: Peronosporaceae) in Flowering Dogwoods (Cornalaes: Cornaceae) After Simulated Flooding.

Ambrosia beetles (Coleoptera: Curculionidae, Scolytinae) and Phytophthora root rot (Peronosporales: Peronosporaceae) cause significant damage to the ornamental industry in the United States. In this study, mefenoxam (fungicide), permethrin (insecticide), and charcoal + kaolin were used in different combinations with Phytophthora cinnamomi (Rands: Peronosporales: Peronosporaceae) inoculated and noninoculated plants to optimize the management of ambrosia beetles and Phytophthora root rot. Treatment applications were performed in two trials on 1 (mefenoxam, drench), 18 (P. cinnamomi inoculation), or 19 (permethrin, spray) days before instigating flood stress or 2 d after flood stress (charcoal + kaolin, spray), respectively. Flooding was maintained for 21 d. Ambrosia beetle attacks and plant growth data were recorded. Tree roots were rated at study end for disease severity and root samples were plated on PARPH-V8 medium to determine the percentage of pathogen recovery. In both trials, the combination of mefenoxam + permethrin treatment had reduced disease severity and ambrosia beetle attacks compared to the inoculated controls. Permethrin-treated trees had shorter galleries compared to controls in trial 1 and no gallery formation in trial 2. In both trials, no differences were observed among the treatments in numbers of galleries with eggs and adults, but mefenoxam + charcoal + kaolin had significantly fewer galleries with larvae among the noninoculated trees compared with the respective control in trial 1. Overall, treatments containing combinations of mefenoxam + permethrin had reduced disease severity and ambrosia beetle attacks.

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

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

Optimizing ethanol-baited traps for monitoring damaging ambrosia beetles (Coleoptera: Curculionidae, Scolytinae) in ornamental nurseries.

The exotic ambrosia beetles Xylosandrus crassiusculus (Motschulsky) and Xylosandrus germanus (Blandford) (Coleoptera: Curculionidae: Scolytinae) are serious pests in ornamental tree nurseries. To optimize bottle-traps as a monitoring system for X. crassiusculus and X. germanus in nurseries, we tested whether increasing the rate of commercial ethanol lures improved captures or early detection of these species. Experiments were conducted in Ohio (2008 and 2009) and Virginia (2008), two states that have experienced significant damage from X. crassiusculus, X. germanus, or both. There were four treatments: no-lure (unbaited control), 1-ethanol lure, 2-ethanol lures and 1 + 1-ethanol lures (one lure in the trap and one suspended 0.5 m above the trap). Captures of X. crassiusculus and X. germanus were higher in all ethanol treatments than unbaited controls, and were generally higher in treatments with two lures versus one. There was no difference in beetle captures between the 2-lure and 1 + 1-lure treatments. First detection of X. crassiusculus and X. germanus occurred more consistently in the treatments with two lures than one lure. Xyleborinus saxesenii (Ratzeburg), Anisandrus sayi Hopkins, Hypothenemus dissimilis Zimmermann, and Hypothenemus eruditus Westwood were also more attracted to traps baited with ethanol than unbaited controls. X. saxesenii was captured in higher numbers in the treatments with two lures than one in Virginia but not in Ohio. There was no difference in captures of the other species among ethanol treatments. The current research shows that ethanol release rates influence sensitivity of traps for detecting emergence of overwintered ambrosia beetles.

Species dependent influence of (-)-alpha-pinene on attraction of ambrosia beetles (Coleoptera: Curculionidae: Scolytinae) to ethanol-baited traps in nursery agroecosystems.

Field-based trapping experiments were conducted in Ohio in 2003, 2004, and 2008 to determine the influence of (-)-alpha-pinene on the attraction of exotic and native ambrosia beetles (Coleoptera: Curculionidae: Scolytinae) to ethanol-baited traps. In 2003 and 2004, we determined the effect of adding an (-)-alpha-pinene ultrahigh release lure (UHR; 2 g/d at 20 degrees C) to traps baited with an ethanol UHR lure (0.39 g/d). FewerAnisandrus (Xyleborus) sayi (Hopkins) and Xyleborinus saxeseni (Ratzeburg) were collected in 2003 and 2004 from traps baited with ethanol UHR plus (-)-alpha-pinene UHR compared with ethanol UHR. (-)-alpha-Pinene also reduced the attraction of Xyloterinus politus (Say) to ethanol-baited traps in 2004. Total captures of Xylosandrus germanus (Blandford) in 2003 were higher in traps baited with ethanol UHR plus (-)-alpha-pinene UHR than in traps with ethanol UHR alone but not in 2004. In 2008, captures were compared among traps baited with eight combinations of ethanol and (-)-a-pinene at both UHR and low release (LR) rates. Release rates for ethanol LR and (-)-alpha-pinene LR were 0.027 and 0.0015 g/d, respectively. (-)-alpha-Pinene UHR and (-)-alpha-pinene LR reduced the attractiveness of ethanol UHR to A. sayi and X. saxeseni. Ethanol UHR was also more attractive than ethanol LR to A. sayi and X. germanus. These findings demonstrate traps baited with ethanol alone are more effective than ethanol plus (-)-alpha-pinene for monitoring ambrosia beetle flight activity in ornamental nurseries. Ethanol release rate is also an important consideration for monitoring purposes.

Comparative efficacy of plant-derived essential oils for managing ambrosia beetles (Coleoptera: Curculionidae: Scolytinae) and their corresponding mass spectral characterization.

Ambrosia beetles (Coleoptera: Curculionidae: Scolytinae) pose a significant challenge to producers of ornamental nursery stock. Conventional insecticides are commonly used for management purposes, but plant-derived essential oils also may discourage ambrosia beetles from initiating attacks. To identify promising commercially available products, field-based efficacy trials were conducted in Ohio in 2009 and 2010 with the following products: Armorex (Soil Technologies), Cinnacure (Proguard, Inc.), EcoTrol (EcoSMART Technologies, Inc.), and Veggie Pharm (Pharm Solutions, Inc.). Potted Magnolia virginiana L. were first injected with 75 ml of 5% ethanol to ensure ambrosia beetle pressure on experimental trees. Mixtures of each product (10% in water) and a water control were applied until runoff and attacks occurring under field conditions were quantified at 1, 4, 7, and 14 d after treatment (DAT). Ambrosia beetle attacks generally increased over time but at differing rates depending on the particular treatment. In 2009, Armorex and Veggie Pharm were associated with the lowest cumulative attacks 14 DAT. In 2010, Armorex and Cinnacure were associated with the fewest attacks 14 DAT. Solid phase microextraction-gas chromatography-mass spectrometry was used to characterize the volatile compounds associated with each product. Allyl isothiocyanate, a compound with known repellent and insecticidal properties, was unique and predominant in Armorex. These experiments identified commercially available botanicals containing plant essential oils with activity against ambrosia beetles, along with demonstrating the usefulness of ethanol-injection to ensure ambrosia beetle pressure under field conditions. Characterizing the constituents of efficacious botanically based products could also lead to the development of improved botanical insecticides.

Unique venom proteins from Solenopsis invicta x Solenopsis richteri hybrid fire ants.

The Solenopsis venom protein 2 transcript was amplified, sequenced, probed, and analyzed from Solenopsis invicta x Solenopsis richteri hybrid ant colonies (hybrids) collected from across Tennessee to determine the extent of introgression of each parent allele (Solenopsis invicta venom protein 2 [Soli2] and Solenopsis richteri venom protein 2 [Solr2]). Chemotaxonomic analyses of venom alkaloids and cuticular hydrocarbons were used to categorize hybrid colonies and their relative relatedness to each parent species. Hybrid colonies were chosen randomly from each chemotaxonomic hybridization category, including "very near S. richteri," "near S. richteri," "near S. invicta," and "very near S. invicta." Lateral flow immunoassays for detection of the Soli2 and Solr2 venom proteins were largely in agreement with the chemotaxonomic analyses for the very near S. richteri (100% Solr2) and very near S. invicta (80% Soli2, 20% Soli2 + Solr2 detected in the sample) groups, while Soli2 and Solr2 were reported in 60% and 40% in the near S. invicta and near S. richteri chemotaxonomic groups. Analysis of transcripts from the hybrid colonies revealed a sequence with 100% identity to Soli2 (GenBank Accession L09560) and three unique sequences, which we identify as Solenopsis hybrid venom protein 2 (Solh2; GenBank Accession MT150127), Solenopsis hybrid truncated venom protein 2 (Solh2Tr97; Genbank Accession MT150129), and Solenopsis richteri venom protein 2, D to A change at position 69 (Solr2A69; GenBank Accession MT150128). The predicted open reading frame for Solh2 and Solh2Tr97 revealed sequences unique to hybrid ants, with Solh2Tr97an alternatively spliced form. A third unique sequence, Solr2A69, is likely the correct sequence for Solr2, which appears to have been published previously with a sequencing error (GenBank Accession P35776).

Evaluation of Systemic Imidacloprid and Herbicide Treatments on Flatheaded Borer (Coleoptera: Buprestidae) Management in Field Nursery Production.

The flatheaded appletree borer, Chrysobothris femorata (Olivier) (Coleoptera: Buprestidae), and related species are deciduous tree pests. Female beetles prefer to oviposit at tree bases, and larvae tunnel beneath the bark, which weakens or kills young or newly transplanted trees. In the first objective of this study, Discus N/G (2.94% imidacloprid + 0.7% cyfluthrin) applied at six lower-than-labeled rates (0.0, 0.98, 1.97, 3.94, 5.91, and 7.87 ml/cm of average trunk dia.) was evaluated for protection of field-grown maples. A second objective evaluated imidacloprid with and without herbicides to assess the impact of weed competition at the tree base on insecticide effectiveness. A third objective determined relative imidacloprid concentrations in leaf tissue samples with ELISA and related to insecticide rates, herbicide treatments, and the level of flatheaded borer protection. In two trials, higher rates of insecticide were more effective at protecting trees, with rates ≥3.94 ml product/cm trunk diameter performing equivalently. Weed-free trees had more borer attacks and grew faster than trees in weedy plots. Imidacloprid content in leaf tissues had a trend for higher concentrations in smaller, weedy trees in the first season, but that pattern disappeared in subsequent years. Based on fewer attacks in weedy versus weed-free trees (60-90% reduction), it was concluded that weed presence can reduce borer attack success in nurseries independent of insecticide treatment, but tree growth was reduced by weed presence. In addition, Discus applied at rates >3.94 ml/cm did not confer added borer damage protection in weedy plots.

Permethrin Residual Activity Against Ambrosia Beetle (Coleoptera: Curculionidae: Scolytinae) Attacks Following Field Aging and Simulated Rainfall Weathering.

Adult ambrosia beetles (Coleoptera: Curculionidae: Scolytinae) bore into ornamental nursery trees resulting in trunk vascular tissue damage, which can potentially kill trees. Ambrosia beetle exposure to surface-applied insecticides is minimal after internal trunk galleries are formed, so effective management requires insecticide treatments to be applied near the time of infestation or to have residual activity on the bark. Tree trunk sections (bolts) were used to determine the effect of field aging or irrigation (i.e., simulated rainfall weathering) on permethrin residual activity against ambrosia beetles. In all experiments, 30-cm-long bolts from Liriodendron tulipifera L. (Magnoliales: Magnoliaceae) were hollowed and filled with 70% ethanol at field deployment to induce ambrosia beetle attacks over a 2-wk period. To evaluate insecticide residual activity, permethrin was sprayed onto tree bolts at 0, 8, 17, or 24 d before ethanol addition, and then bolts were deployed along a wooded border in fall 2017 and spring 2018. Tree bolts with permethrin residues ≤17 d old had significantly fewer ambrosia beetle attacks than bolts with 24-d-old residues or the non-permethrin-treated control bolts. To evaluate simulated rainfall weathering, permethrin was applied to tree bolts 8 or 22 d before ethanol (spring 2018) or 10 or 24 d before ethanol (fall 2018) with half of the bolts receiving regular irrigation events. Irrigation had no significant effect on permethrin residual activity against ambrosia beetles during either test. This study determined ambrosia beetle control was affected by permethrin residue age more than simulated rainfall weathering, and a reapplication interval of ≤17 d maximized beetle control.

Surface-applied insecticide treatments for quarantine control of Japanese beetle, Popillia japonica Newman (Coleoptera: Scarabaeidae), larvae in field-grown nursery plants.

BACKGROUND: Japanese beetles, Popillia japonica Newman, are a quarantine challenge for nursery shipments from infested to non-infested states. Marathon (imidacloprid) and Discus (imidacloprid + cyfluthrin) are approved preharvest nursery treatments (US Domestic Japanese Beetle Harmonization Plan; DJHP). This study evaluated approved and non-approved (acephate, carbaryl, clothianidin, dinotefuran, halofenozide, thiamethoxam, trichlorfon) preharvest treatments, optimal rates (labeled 1x, 0.3-0.75 x or 2-3 x) and optimal timings (June, July, August and September) to control early-instar (grubs) P. japonica in field nurseries. RESULTS: Most insecticides effectively reduced grub densities, except for acephate, carbaryl and trichlorfon. Clothianidin, thiamethoxam and halofenozide provided grub control equivalent to DJHP standards during most years. Across all test years and timings, percentage grub reductions were: Marathon (1x: 59.2-100; 3 x: 78.9-100), Discus (1x: 60.7-100), clothianidin (1x: 96.1-100; 3 x: 97.4-100), thiamethoxam (1x: 75.0-100; 3 x: 80.0-100), halofenozide (1x: 70.0-100; 3 x: 90.0-100) and dinotefuran (1x: 13.2-88.2; 3 x: 71.1-93.4). CONCLUSIONS: July application was more consistent and effective than other timings, and higher rates (2x and 3 x) did not generally improve grub control. Overall, clothianidin, thiamethoxam and halofenozide (and dinotefuran applied in August) were equivalent to current DJHP standards. These insecticides may be suitable for DJHP Category 2 states, potentially lowering grower costs.

Toxicity of botanical formulations to nursery-infesting white grubs (Coleoptera: Scarabaeidae).

The toxicity of eight botanically based biopesticides was evaluated against third instars of the scarab larvae (Coleoptera: Scarabaeidae) Popillia japonica Newman, Rhizotrogus majalis (Razoumowsky), Anomala orientalis Waterhouse, and Cyclocephala borealis Arrow. Soil dip bioassays were used to obtain concentration-mortality data 7 d after treatment of larvae, leading to the calculation of LC50 and LC90 values. A wide range in LC50 and LC90 values were exhibited among the formulations. The product Armorex was one of the most active formulations against P. japonica (LC50 = 0.42 ml/liter), R. majalis (LC50 = 0.48 ml/liter), A. orientalis (LC50 = 0.39 ml/liter), and C. borealis (LC50 = 0.49 ml/liter). Armorex is composed of extracts from diverse botanical sources, including 84.5% sesame oil, 2.0% garlic oil, 2.0% clove oil, 1.0% rosemary oil, and 0.5% white pepper extracts. The product Azatin, composed of 3% azadirachtin, also exhibited high toxicity to P. japonica (LC50 = 1.13 ml/liter), R. majalis (LC50 = 0.81 ml/liter), and A. orientalis (LC50 = 1.87 ml/liter). Veggie Pharm is composed of extracts from diverse sources, but this product showed the lowest toxicity to P. japonica (LC50 = 35.19 ml/liter), R. majalis (LC50 = 62.10 ml/liter), A. orientalis (LC50 = 43.76 ml/liter), and C. borealis (LC50 = 50.24 ml/liter). These results document the potential for botanical formulations to control white grubs, but blending extracts from diverse botanical sources does not ensure enhanced biological activity.