Bacterial microbiome associated with cigarette beetle Lasioderma serricorne (F.) and its microbial plasticity in relation to diet sources.

Insect-microbial symbiosis contributes positively to the physiology of the insect and diet is considered as one important factor determining microbial symbiosis. In this study, we have characterized the microbiota of cigarette beetle, Lasioderma serricorne (Fabricius) on different diets and phases. The beetles were reared on different diet sources (exposed phase) for six generations and were reverted to their natal source (reverted phase) and further maintained for six more generations. The bacterial diversity and richness were higher in the exposed phase and once reverted to the natal source, the microbial abundance has re-assembled according to the natal diet source. There was re-assemblage of microbial composition in accordance to the diet and the bacterial cells are able to establish and proliferate on reverting to their natal source. The bacterial composition of the beetle was mainly dynamic and not transient where the bacterial cells were maintained at low abundance and were re-established according to the diet source. Overall, we found that the microbiota of cigarette beetle to be dynamic and bacterial composition to re-assemble in a diet-specific manner. The study provides insights on diet associated microbial plasticity of cigarette beetle and a further comprehensive understanding on mechanisms involved in microbial plasticity will help develop novel pest management strategies for this invasive insect pest.

Distribution and accumulation of Cadmium in different trophic levels affecting Serangium japonicum, the predatory beetle of whitefly Bemisia tabaci, biologically, physiologically and genetically: An experimental study.

Cadmium (Cd) is a heavy metal that is of great concern in agroecosystems due to its toxicity to plants, herbivores, carnivores, and human beings. The current study evaluated the allocation and bioaccumulation of Cd from soil to cotton plants, cotton plants to herbivore pests, and herbivorous pests to a natural enemy predator. When soil was spiked with 100 mg/kg Cd, results demonstrated that cotton roots accumulated more Cd than the stems and leaves. The bioaccumulation of Cd was less in 4th instar larvae, pupa, and adults of Serangium japonicum than in Bemisia tabaci adults. The bioaccumulation in S. japonicum elongated the immature development period and reduced adult longevity, oviposition days, fertility, and total pre-oviposition duration. The net reproduction of S. japonicum was also reduced, as was female mature weight and feeding potential; as a result, Cd exposure could reduce the future population size compared to uncontaminated populations. There was decreased activity of the antioxidant enzymes (SOD, CAT, and POD) and energy-conserving lipids (glycogen, triglyceride, and total cholesterol) in Cd-contaminated S. japonicum compared to controls. The detoxifying enzyme activity of GST and P450 increased while AChE activity did not change. The qRT-PCR research showed that SOD1, CAT, POD, glycogen, and triglyceride gene expression was higher than in controls, whereas detoxification gene expression did not change. Our results indicate that Cd exposure has a physiological trade-off between its adverse effects on life history traits and elevated detoxification and antioxidation of S. japonicum, which could result from gene expression alteration. Further studies are needed to assess whether Cd exposure causes irreversible DNA damage in S. japonicum.

Assessing the evolutionary lability of insulin signalling in the regulation of nutritional plasticity across traits and species of horned dung beetles.

Nutrition-dependent growth of sexual traits is a major contributor to phenotypic diversity, and a large body of research documents insulin signalling as a major regulator of nutritional plasticity. However, findings across studies raise the possibility that the role of individual components within the insulin signalling pathway diverges in function among traits and taxa. Here, we use RNAi-mediated transcript depletion in the gazelle dung beetle to investigate the functions of forkhead box O (Foxo) and two paralogs of the insulin receptor (InR1 and InR2) in shaping nutritional plasticity in polyphenic male head horns, exaggerated fore legs, and weakly nutrition-responsive genitalia. Our functional genetic manipulations led to three main findings: FoxoRNAi reduced the length of exaggerated head horns in large males, while neither InR1 nor InR2 knock-downs resulted in measurable horn phenotypes. These results are similar to those documented previously for another dung beetle (Onthophagus taurus), but in stark contrast to findings in rhinoceros beetles. Secondly, knockdown of Foxo, InR1, and InR2 led to an increase in the intercept or slope of the scaling relationship of genitalia size. These findings are in contrast even to results documented previously for O. taurus. Lastly, while FoxoRNAi reduces male forelegs in D. gazella and O. taurus, the effects of InR1 and InR2 knockdowns diverged across dung beetle species. Our results add to the growing body of literature indicating that despite insulin signalling's conserved role as a regulator of nutritional plasticity, the functions of its components may diversify among traits and species, potentially fuelling the evolution of scaling relationships.

Tissue-specific plant toxins and adaptation in a specialist root herbivore.

In coevolution between plants and insects, reciprocal selection often leads to phenotype matching between chemical defense and herbivore offense. Nonetheless, it is not well understood whether distinct plant parts are differentially defended and how herbivores adapted to those parts cope with tissue-specific defense. Milkweed plants produce a diversity of cardenolide toxins and specialist herbivores have substitutions in their target enzyme (Na+/K+-ATPase), each playing a central role in milkweed-insect coevolution. The four-eyed milkweed beetle (Tetraopes tetrophthalmus) is an abundant toxin-sequestering herbivore that feeds exclusively on milkweed roots as larvae and less so on milkweed leaves as adults. Accordingly, we tested the tolerance of this beetle's Na+/K+-ATPase to cardenolide extracts from roots versus leaves of its main host (Asclepias syriaca), along with sequestered cardenolides from beetle tissues. We additionally purified and tested the inhibitory activity of dominant cardenolides from roots (syrioside) and leaves (glycosylated aspecioside). Tetraopes' enzyme was threefold more tolerant of root extracts and syrioside than leaf cardenolides. Nonetheless, beetle-sequestered cardenolides were more potent than those in roots, suggesting selective uptake or dependence on compartmentalization of toxins away from the beetle's enzymatic target. Because Tetraopes has two functionally validated amino acid substitutions in its Na+/K+-ATPase compared to the ancestral form in other insects, we compared its cardenolide tolerance to that of wild-type Drosophila and CRISPR-edited Drosophila with Tetraopes' Na+/K+-ATPase genotype. Those two amino acid substitutions accounted for >50% of Tetraopes' enhanced enzymatic tolerance of cardenolides. Thus, milkweed's tissue-specific expression of root toxins is matched by physiological adaptations in its specialist root herbivore.

PI3K/Akt/CncC signaling pathway mediates the response to EPN-Bt infection in Holotrichia parallela larvae.

BACKGROUND: Combining the entomopathogenic nematode (EPN), Heterorhabditis beicherriana LF strain, and Bacillus thuringiensis (Bt) HBF-18 strain is a practical strategy to manage the larvae of Holotrichia parallela Motschulsky (white grubs). However, the mechanisms underlying the larval defense response to this combined biocontrol strategy are unknown. RESULTS: The activities of some antioxidant enzymes (SOD, POD, CAT) and some detoxifying enzymes (AChE, P-450, CarE, GST) in grubs showed an activation-inhibition trend throughout the EPN-Bt exposure time course. Eight potentially key antioxidant and detoxifying enzyme genes in response to EPN-Bt infection were identified from the midgut of grubs through RNA sequencing. After silencing CAT, CarE18, and GSTs1, the enzyme activities were significantly decreased by 30.29%, 68.80%, and 34.63%, respectively. Meanwhile, the mortality of grubs was increased by 18.40%, 46.30%, and 42.59% after exposure to EPN-Bt for 1 day. Interestingly, the PI3K/Akt signaling pathway was significantly enriched in KEGG enrichment analysis, and the expression levels of phosphatidylinositol 3-kinase (PI3K), protein kinase B (Akt), cap 'n' collar isoform-C (CncC), kelch-like ECH-associated protein 1 (Keap1), and CarE18 were all up-regulated when exposed to EPN-Bt for 1 day. Furthermore, RNAi-mediated PI3K silencing showed a similar down-regulated trend between PI3K/Akt/CncC and CarE18. Moreover, silencing PI3K rendered grubs more susceptible to EPN-Bt and accelerated symbiotic bacteria multiplication in grubs. CONCLUSION: These results suggest that the PI3K/Akt/CncC pathway mediates the expression of CarE18 and participates in the defense response of H. parallela larvae against EPN-Bt infection. Our data provide valuable insights into the design of appropriate management strategies for this well-known agricultural pest. © 2022 Society of Chemical Industry.

Evidence for Cardiac Glycosides in Foliage of Colorado Potato Beetle-Resistant Solanum okadae.

Leptinotarsa decemlineata, the Colorado potato beetle (CPB), is a herbivore that primarily feeds on Solanum foliage and is a global pest of the potato agricultural industry. Potato breeding through cross-hybridization with CPB-resistant wild relatives is used for genetic improvement. The wild species Solanum okadae was demonstrated to deter CPB feeding in choice and no choice feeding assays. Liquid chromatography-mass spectrometry (LC-MS) was used for comparative metabolite profiling between S. okadae and CPB-susceptible domesticated potato variety, Solanum tuberosum cv. Shepody. Major foliar metabolites detected were steroidal glycoalkaloids (SGAs) with tomatine and dehydrotomatine produced in S. okadae and solanine and chaconine in S. tuberosum cv. Shepody. Cardiac glycosides were also detected in the foliar metabolite profile of S. okadae but not S. tuberosum cv. Shepody. This class of plant compounds have known insecticidal activity through inhibition of animal Na+/K+ ATPase. Thin-layer chromatography (TLC) separation of foliar extracts also provided evidence for cardiac glycosides in S. okadae. Cardiac glycosides are known inhibitors of Na+/K+ ATPase, and foliar extracts from S. okadae (OKA15), but not S. tuberosum cv. Shepody, were able to inhibit the Na+/K+ ATPase of CPB. These findings suggest a novel mechanism of plant resistance against CPB involving production of cardiac glycosides in S. okadae.

Effect of temperature on the life cycle of Harmonia axyridis (Pallas), and its predation rate on the Spodoptera litura (Fabricius) eggs.

Biological control is one of the strategies of pest control which is determined by the biological fitness and metabolic rates of the predator species used. Temperature and resource are important factors which influence the role of insects as biocontrol agents. Harmonia axyridis is a cosmopolitan and non-specific polyphagous predator. It can survive ecologically diverse environments and exploit multiple preys. This study investigated the effects of temperature on the population parameters of H. axyridis and its predation on the eggs of prey Spodoptera litura. For this purpose, an age-stage, two-sex life table of the predator was constructed at four constant temperatures, i.e. 15, 20, 25 and 30 °C, under laboratory settings of: 70 ± 5% RH, and 16:8 h (L: D) photoperiod. A computer simulation was then used to project the population and predation responses with respect to temperatures tested. We found that the development of larvae and adult (male/female) stages of H. axyridis decreased with colder temperatures (i.e., 15 and 20 °C) but increased with warmer temperatures (25 and 30 °C). The intrinsic rate of increase (r) and mean generation time (T) were 0.0662 d-1 and 79.84 d at 15 °C, 0.0843 d-1 and 64.90 d at 20 °C, 0.1067 d-1 and 48.89 d at 25 °C, and 0.1378 d-1 and 35.55 d at 30 °C, respectively. The mean duration of the total pre-adult stage was 44.26, 32.91, 20.63, and 15.39 d at 15, 20, 25, and 30 °C, respectively. At 30 °C. the finite rate of increase (1.1477 d-1) was the highest and the mean generation time (35.55 d) was the shortest. The net predation rate (C0) was 7935.54, 10,466.28, 10,139.38, and 7126.36 eggs at 15, 20, 25, and 30 °C, respectively. Population and predation projections were proportional to temperature. These findings are important for modelling the population responses of H. axyridis to climate change and tailoring integrated pest management strategies to altered climates.

Comparative proteomic analysis reveals insights into the response of Cryptolaemus montrouzieri to bottom-up transfer of cadmium and lead across a multi-trophic food chain.

Contamination of agro-ecosystems with heavy metals can affect the development and reproduction of insect natural enemies. This study reports a detailed Tandem Mass Tag based quantitative proteomic analysis of underlying mechanisms responsible for stress response of Cryptolaemus montrouzieri against heavy metals (cadmium (Cd) and lead (Pb)) transported across a multi-trophic food chain. A total of 6639 proteins were detected under Cd as well as Pb stress. In Pb versus the control cluster, 69 proteins (28 up-regulated and 41 down-regulated) were differentially expressed whereas 268 proteins were differentially expressed under Cd versus the control cluster, having 198 proteins up-regulated and 70 down-regulated proteins. The analysis of differentially expressed proteins showed that 27 proteins overlapped in both clusters representing the core proteome to Pb and Cd stress. The bioinformatics analysis demonstrated that these proteins were mapped to 57 and 99 pathways in Pb versus control and Cd versus control clusters, respectively. The functional classification by COG, GO and KEGG databases showed significant changes in protein expression by C. montrouzieri under Pb and Cd stress. The heavy metal stress (Pb and Cd) induced significant changes in expression of proteins like hexokinase (HK), succinyl-CoA, trypsin like proteins, cysteine proteases, cell division cycle proteins, and yellow gene proteins. The results provide detailed information on the protein expression levels of C. montrouzieri and will serve as basic information for future proteomic studies on heavy metal responses of insect predators within a multi-trophic food chain.

A robotic leg inspired from an insect leg.

While most insect-inspired robots come with a simple tarsus, such as a hemispherical foot tip, insect legs have complex tarsal structures and claws, which enable them to walk on complex terrain. Their sharp claws can smoothly attach and detach on plant surfaces by actuating a single muscle. Thus, installing an insect-inspired tarsus on legged robots would improve their locomotion on complex terrain. This paper shows that the tendon-driven ball-socket structure provides the tarsus with both flexibility and rigidity, which is necessary for the beetle to walk on a complex substrate such as a mesh surface. Disabling the tarsus' rigidity by removing the socket and elastic membrane of a tarsal joint, means that the claws could not attach to the mesh securely. Meanwhile, the beetle struggled to draw the claws out of the substrate when we turned the tarsus rigid by tubing. We then developed a cable-driven bio-inspired tarsus structure to validate the function of the tarsus as well as to show its potential application in the legged robot. With the tarsus, the robotic leg was able to attach and retract smoothly from the mesh substrate when performing a walking cycle.

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.

Continuous phenotypic modulation explains male horn allometry in three dung beetle species.

Many dung beetle species show male horn polyphenism, the ability of males to develop into distinct phenotypes without intermediate forms as a response to the larval growth environment. While males with long (majors) and rudimentary (minor) horn have been widely reported in literature, little is known about the existence of individuals with intermediate horn length. Here we investigate the occurrence of intermediates in natural populations of three dung beetle species (Onthophagus furcatus, Copris lunaris and C. hispanus). We analysed the body size-horn length relationship using linear, exponential, and sigmoidal models with different error structures. We inferred the number of individuals in the minor, intermediate, and major groups by combining changepoint analysis and simulation from fitted allometric models. The sigmoidal equation was a better descriptor of the body size-horn length relationship than linear or exponential equations in all the three studied species. Our results indicated that the number of intermediates equals or exceeds the number of minor and major males. This work provides evidence that, at least in the studied species, males with intermediate horn length exist in natural populations. For similar cases we therefore suggest that continuous phenotypic modulation rather than discrete polyphenism can explain variation in male horn allometry.

Comparative metabolomic analysis of polyphenic horn development in the dung beetle Onthophagus taurus.

Organisms alter their phenotypes in response to changing environmental conditions. The developmental basis of this phenomenon, phenotypic plasticity, is a topic of broad interest in many fields of biology. While insects provide a suitable model for studying the genetic basis of phenotypic plasticity, the physiological aspects of plasticity are not fully understood. Here, we report the physiological basis of polyphenism, an extreme form of phenotypic plasticity by utilizing a dung beetle species, Onthophagus taurus. We highlighted the metabolome between sexes as well as two distinct male morphs-large and small horns. Unlike results from previous transcriptomic studies, the comparative metabolomic study revealed that differences in metabolite level were more prominent between animals with different body sizes than different sexes. Our results also indicate that specific metabolites and biochemical pathways may be active during horn size determination.

Biochemical and histological alterations induced by nickel oxide nanoparticles in the ground beetle Blaps polychresta (Forskl, 1775) (Coleoptera: Tenebrionidae).

The present study evaluates the effect of nickel oxide nanoparticles on some biochemical parameters and midgut tissues in the ground beetle Blaps polychresta as an indicator organism for nanotoxicity. Serial doses of the NiO-NPs colloid (0.01, 0.02, 0.03, 0.04, 0.05, and 0.06 mg/g) were prepared for injecting into the adult beetles. Insect survival was reported daily for 30 days, and the sublethal dose of 0.02 mg/g NiO-NPs was selected for the tested parameters. After the treatment, nickel was detected in the midgut tissues by X-ray microanalysis. The treated group demonstrated a significant increase in aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities when compared to the untreated group. However, the treated group demonstrated a significant decrease in ascorbate peroxidase (APOX) activity when compared to the untreated group. Histological and ultrastructural changes in the midgut tissues of treated and untreated beetles were also observed. The current findings provide a precedent for describing the physiological and histological changes caused by NiO-NPs in the ground beetle B. polychresta.

Total-evidence backbone phylogeny of Aleocharinae (Coleoptera: Staphylinidae).

Phylogenetic studies of Aleocharinae rove beetles, arguably one of the least known and the largest insect lineages, are compromised by its enormous taxonomic diversity. DNA, a powerful resource for phylogenetics, is not available for numerous extant aleocharine species. We provide a broad comparative morphological study of Aleocharinae to frame molecular datasets for total-evidence analyses. Using full-body dissections and slide-mounting techniques for light microscopy supplemented by scanning electron microscopy, we constructed a morphological matrix across all major taxa focused on non-inquiline tribes of Aleocharinae and outgroups. Phylogenetic analyses of this matrix concatenated with earlier published DNA loci and including exemplar taxa lacking molecular data, resolved outstanding controversies and, among other novelties, showed that: the Habrocerinae + Trichophyinae clade is sister group to Aleocharinae; Hypocyphtini are sister to the rest of the "higher Aleocharinae"; Taxicerini are sister to Aleocharini; Hoplandriini and Placusini are nested within a polyphyletic Oxypodini; Hoplandriini are sister to Meoticina; and Actocharini are nested within Liparocephalini. For the first time, morphological synapomorphies are identified for some large clades of Aleocharinae. In addition, 1252 high-resolution microphotographs of aleocharine structures are made available online with the entire matrix for future research.

Gas exchange and dive behaviour in the diving beetle Platynectes decempunctatus (Coleoptera: Dytiscidae).

Many aquatic insects use bubbles on the body surface to store and supply O2 for their dives. There are two types of bubbles: air stores, which store O2 gained from air at the surface, and gas gills that allow passive extraction of O2 from water. Many insects using air stores and gas gills return to the surface to replenish their bubbles and, therefore, their requirement for O2 influences dive behaviour. In this study, we investigate gas exchange and dive behaviour in the diving beetle Platynectes decempunctatus that uses a sub-elytral air store and a small compressible gas gill. We measure the PO2 within the air store during tethered dives, as well as the amount of O2 exchanged during surfacing events. Buoyancy experiments monitor the volume of gas in the gas gill and how it changes during dives. We also directly link O2-consumption rate at three temperatures (10, 15 and 20 °C) with dive duration, surfacing frequency and movement activity. These data are incorporated in a gas exchange model, which shows that the small gas gill of P. decempunctatus contributes less than 10% of the total O2 used during the dive, while up to 10% is supplied by cutaneous uptake.

Thermal radiation management by natural photonic structures: Morimus asper funereus case.

Convective, conductive and radiative mechanisms of thermal management are extremely important for life. Photonic structures, used to detect infrared radiation (IR) and enhance radiative energy exchange, were observed in a number of organisms. Here we report on sophisticated radiative mechanisms used by Morimus asper funereus, a longicorn beetle whose elytra possess a suitably aligned array of lenslets and blackbodies. Additionally, a dense array of microtrichia hyperuniformly covers blackbodies and operates as a stochastic, full-bandgap, IR-photonic structure. All these features, whose characteristic dimensions cover a range from several hundred down to a few micrometres, operate synergistically to improve the absorption, emission and, possibly, detection of IR radiation. We present a morphological characterization of the elytron, thermal imaging measurements and a theoretical IR model of insect elytron, uncovering a synergistic operation of all structures.

Climate change impact on the potential geographical distribution of two invading Xylosandrus ambrosia beetles.

Xylosandrus compactus and X. crassiusculus are two polyphagous ambrosia beetles originating from Asia and invasive in circumtropical regions worldwide. Both species were recently reported in Italy and further invaded several other European countries in the following years. We used the MaxEnt algorithm to estimate the suitable areas worldwide for both species under the current climate. We also made future projections for years 2050 and 2070 using 11 different General Circulation Models, for 4 Representative Concentration Pathways (2.6, 4.5, 6.0 and 8.5). Our analyses showed that X. compactus has not been reported in all potentially suitable areas yet. Its current distribution in Europe is localised, whereas our results predicted that most of the periphery of the Mediterranean Sea and most of the Atlantic coast of France could be suitable. Outside Europe, our results also predicted Central America, all islands in Southeast Asia and some Oceanian coasts as suitable. Even though our results when modelling its potential distribution under future climates were more variable, the models predicted an increase in suitability poleward and more uncertainty in the circumtropical regions. For X. crassiusculus, the same method only yielded poor results, and the models thus could not be used for predictions. We discuss here these results and propose advice about risk prevention and invasion management of both species.

Remote radio control of insect flight reveals why beetles lift their legs in flight while other insects tightly fold.

In the research and development of micro air vehicles, understanding and imitating the flight mechanism of insects presents a viable way of progressing forward. While research is being conducted on the flight mechanism of insects such as flies and dragonflies, research on beetles that can carry larger loads is limited. Here, we clarified the beetle midlegs' role in the attenuation and cessation of the wingbeat. We anatomically confirmed the connection between the midlegs and the elytra. We also further clarified which pair of legs are involved in the wingbeat attenuation mechanism, and lastly demonstrated free-flight control via remote leg muscle stimulation. Observation of multiple landings using a high-speed camera revealed that the wingbeat stopped immediately after their midlegs were lowered. Moreover, the action of lowering the midleg attenuated and often stopped the wingbeat. A miniature remote stimulation device (backpack) mountable on beetles was designed and utilized for the free-flight demonstration. Beetles in free flight were remotely induced into lowering (swing down) each leg pair via electrical stimulation, and they were found to lose significant altitude only when the midlegs were stimulated. Thus, the results of this study revealed that swinging down of the midlegs played a significant role in beetle wingbeat cessation. In the future, our findings on the wingbeat attenuation and cessation mechanism are expected to be helpful in designing bioinspired micro air vehicles.

Dormancy in laboratory-reared Asian longhorned beetles, Anoplophora glabripennis.

An insect's capacity to survive winter is critical for range expansion in temperate regions. The Asian longhorned beetle (Anoplophora glabripennis) is a polyphagous wood-boring insect native to China and the Korean peninsula and poses a high risk of invasion in North America and Europe. It is unclear whether A. glabripennis enters diapause, which means that diapause cannot be included in assessments of the risk of this species invading forests in temperate regions. Using a laboratory colony, we examine larval developmental arrest, metabolic rates, gas exchange patterns, thermal sensitivity, and body composition to characterize larval dormancy. Chilled larvae entered a temperature-independent developmental arrest which usually required more than four weeks of chilling to break, decreased their metabolic rate by as much as 63%, and maintained energy stores throughout the chilling period - results consistent with an obligate diapause. We also observed a switch to discontinuous gas exchange at low temperatures. Thermal sensitivity of metabolic rate did not differ between chilled and non-chilled larvae. Taken together, we conclude that A. glabripennis enters a larval diapause during chilling and terminates diapause after a requisite chilling period. These results will enhance our ability to predict phenology and potential distribution of current and future invasions of A. glabripennis.

Light Limitation Impacts Growth but Not Constitutive or Jasmonate Induced Defenses Relevant to Emerald Ash Borer (Agrilus planipennis) in White Fringetree (Chionanthus virginicus) or Black Ash (Fraxinus nigra).

White fringetree is a host for the invasive emerald ash borer (EAB) but is of lower quality than the related and highly susceptible black ash. Field observations suggest that host trees grown in full sun are more resistant to EAB than those in shade, however the impact of light limitation on chemical defenses has not been assessed. We quantified constitutive and jasmonate-induced phloem defenses and growth patterns of white fringetree and black ash under differential light conditions and related them to EAB larval performance. White fringetree had significantly lower constitutive and induced activities of peroxidase, polyphenol oxidase, ß-glucosidase, chitinase and lignin content, but significantly higher gallic acid equivalent soluble phenolic, soluble sugar, and oleuropein concentrations compared to black ash. Multivariate analyses based on tissue chemical attributes displayed clear separation of species and induced defense responses. Further, EAB performed significantly worse on white fringetree than black ash, consistent with previous studies. Light limitation did not impact measured defenses or EAB larval performance, but it did decrease current year growth and increase photosynthetic efficiency. Overall our results suggest that phenolic profiles, metabolite abundance, and growth traits are important in mediating white fringetree resistance to EAB, and that short-term light limitation does not influence phloem chemistry or larval success.