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Herbivores are commonly classified as host specialists or generalists for various purposes, yet the definitions of these terms, and their intermediates, are often imprecise and ambiguous. We quantified host breadth for 240 non-native, tree-feeding insects in North America using phylogenetic diversity. We demonstrated that a partitioning of host breadth: (1) causes 67% of non-native insects to shift from a generalist to specialist category, (2) displays a reduction in host breadth from the native to introduced range, (3) identifies an inflection point in a model predicting the likelihood of non-native insect ecological impact, with a corresponding change in behaviour associated with specialists versus generalists, and (4) enables three models for strong prediction of whether a non-native forest insect will cause high impacts. Together, these results highlight the primacy of how herbivore host recognition and plant defences mediate whether novel host interactions will result in high impact after invasion., (© 2025 John Wiley & Sons Ltd. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.)

Some introduced species cause severe damage, although the majority have little impact. Robust predictions of which species are most likely to cause substantial impacts could focus efforts to mitigate those impacts or prevent certain invasions entirely. Introduced herbivorous insects can reduce crop yield, fundamentally alter natural and managed forest ecosystems, and are unique among invasive species in that they require certain host plants to succeed. Recent studies have demonstrated that understanding the evolutionary history of introduced herbivores and their host plants can provide robust predictions of impact. Specifically, divergence times between hosts in the native and introduced ranges of a nonnative insect can be used to predict the potential impact of the insect should it establish in a novel ecosystem. However, divergence time estimates vary among published phylogenetic datasets, making it crucial to understand if and how the choice of phylogeny affects prediction of impact. Here, we tested the robustness of impact prediction to variation in host phylogeny by using insects that feed on conifers and predicting the likelihood of high impact using four different published phylogenies. Our analyses ranked 62 insects that are not established in North America and 47 North American conifer species according to overall risk and vulnerability, respectively. We found that results were robust to the choice of phylogeny. Although published vascular plant phylogenies continue to be refined, our analysis indicates that those differences are not substantial enough to alter the predictions of invader impact. Our results can assist in focusing biosecurity programs for conifer pests and can be more generally applied to nonnative insects and their potential hosts by prioritizing surveillance for those insects most likely to be damaging invaders., (© 2022 The Ecological Society of America. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.)

Emerald ash borer ( Agrilus planipennis ; EAB) has devastated populations of ash ( Fraxinus spp.) trees in dozens of U.S. states and Canada over the past few decades. The continued survival of scattered ash trees known as "lingering ash" in heavily infested natural stands, however, offers evidence of genetic resistance or tolerance to EAB. These surviving or "lingering" ash individuals may form the basis for reforestation programs in EAB-impacted areas, and clonal mass-propagation of these genotypes can help accelerate these efforts. Between 2013 and 2018, we initiated embryogenic cultures by culturing immature zygotic embryos from open-pollinated (OP) seeds collected from several surviving white ash and green ash trees in Michigan and Pennsylvania. In addition, in 2018, we initiated cultures from crosses made between lingering green ash parents from the USDA Forest Service ash breeding program in Ohio. Somatic embryos were produced by growing cultures in liquid suspension, followed by fractionation and plating on semisolid medium to produce developmentally synchronous populations of somatic embryos. Somatic embryo germination and conversion were enhanced by a combination of pre-germination cold treatment and inclusion of activated charcoal and gibberellic acid in the germination medium. Ash somatic seedlings derived from OP explants grew rapidly following transfer to potting mix and somatic seedlings representing nine ash clones were acclimatized, grown in the greenhouse and planted in a preliminary field test, along with EAB-resistant Manchurian ash ( F. mandshurica ) and EAB-susceptible control seedlings. Somatic seedlings have now been produced from cultures that originated from seeds derived from the progeny of lingering green ash parents and an ex vitro germination protocol has shown some promise for accelerating early somatic seedling growth. Results of this research could provide the basis for scaled-up production of EAB-resistant ash varieties for seed orchard production for forest restoration and cultivar development for urban tree restoration., Competing Interests: Conflict of interestThe authors declare that they have no conflicts of interest., (© The Author(s), under exclusive licence to Springer Nature B.V. 2022.)

Management responses to invasive forest insects are facilitated by the use of detection traps ideally baited with species-specific semiochemicals. Emerald ash borer, Agrilus planipennis Fairmaire, is currently invading North American forests, and since its detection in 2002, development of monitoring tools has been a primary research objective. We compared six trapping schemes for A. planipennis over 2 yr at sites in four U.S. states and one Canadian province that represented a range of background A. planipennis densities, canopy coverage, and ash basal area. We also developed a region-wide phenology model. Across all sites and both years, the 10th, 50th, and 90th percentile of adult flight occurred at 428, 587, and 837 accumulated degree-days, respectively, using a base temperature threshold of 10°C and a start date of 1 January. Most trapping schemes captured comparable numbers of beetles with the exception of purple prism traps (USDA APHIS PPQ), which captured significantly fewer adults. Trapping schemes varied in their trap catch across the gradient of ash basal area, although when considering trap catch as a binary response variable, trapping schemes were more likely to detect A. planipennis in areas with a higher ash component. Results could assist managers in optimizing trap selection, placement, and timing of deployment given local weather conditions, forest composition, and A. planipennis density., (© The Author(s) 2021. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

A long-standing goal of invasion biology is to identify factors driving highly variable impacts of non-native species. Although hypotheses exist that emphasize the role of evolutionary history (e.g., enemy release hypothesis & defense-free space hypothesis), predicting the impact of non-native herbivorous insects has eluded scientists for over a century.Using a census of all 58 non-native conifer-specialist insects in North America, we quantified the contribution of over 25 factors that could affect the impact they have on their novel hosts, including insect traits (fecundity, voltinism, native range, etc.), host traits (shade tolerance, growth rate, wood density, etc.), and evolutionary relationships (between native and novel hosts and insects).We discovered that divergence times between native and novel hosts, the shade and drought tolerance of the novel host, and the presence of a coevolved congener on a shared host, were more predictive of impact than the traits of the invading insect. These factors built upon each other to strengthen our ability to predict the risk of a non-native insect becoming invasive. This research is the first to empirically support historically assumed hypotheses about the importance of evolutionary history as a major driver of impact of non-native herbivorous insects.Our novel, integrated model predicts whether a non-native insect not yet present in North America will have a one in 6.5 to a one in 2,858 chance of causing widespread mortality of a conifer species if established ( R 2  = 0.91) Synthesis and applications . With this advancement, the risk to other conifer host species and regions can be assessed, and regulatory and pest management efforts can be more efficiently prioritized., Competing Interests: The authors declare no conflict of interest., (© 2019 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.)

Phenological synchrony between herbivorous insects and host plants is an important determinant of insect distribution and abundance. Non-native insects often experience novel climates, photoperiods, and host plants. How critical time periods of insect life cycles coincide with-or diverge from-phenological windows of host plant suitability could affect invasion success and the dynamics of outbreaks. Larch casebearer is an invasive defoliator that has recently undergone anomalous outbreaks on eastern larch in North America. We conducted growth chamber, greenhouse, and field studies to quantify the spring phenological window for larch casebearer on eastern larch and importance of phenological synchrony for casebearer development and survival. We constructed degree-day models of spring activity for both species and investigated responses of casebearers to early and delayed activation relative to bud break. Both species had lower developmental thresholds of  ~ 5 °C, but mean activation of casebearers occurred 245 degree-days after bud break by eastern larch. In addition to forcing temperatures, phenologies of eastern larch and casebearer larvae were significantly influenced by chilling and photoperiod, respectively. Larvae were robust to both starvation and delayed activation; days between larval activation and bud break (range: 0-58 days) had no influence on larval development and survival to adulthood. Disparate plant-insect responses to environmental cues and robustness of casebearers to changes in phenology result in a wide phenological window that likely has contributed to the insect's broad distribution in eastern North America. Changes in phenological synchrony, however, do not appear to have facilitated recent outbreaks of larch casebearer on eastern larch.

In forest ecosystems, natural and anthropogenic disturbances alter canopy structure, understory vegetation, amount of woody debris, and the properties of litter and soil layers. The magnitude of these environmental changes is context-dependent and determined by the properties of the disturbance, such as the frequency, intensity, duration, and extent. Therefore, disturbances can dynamically impact forest communities over time, including populations of ground-dwelling invertebrates that regulate key ecosystem processes. We propose conceptual models that describe the dynamic temporal effects of canopy gap formation and coarse woody debris accumulation following disturbances caused by invasive insects, wind, and salvage logging, and their impacts on ground-dwelling invertebrate communities. Within this framework, predictions are generated, literature on ground-dwelling invertebrate communities is synthesized, and pertinent knowledge gaps identified.

Emerald ash borer (EAB; Agrilus planipennis) is a devastating pest of ash (Fraxinus spp.) in its invaded range in North America. Its coevolved Asian hosts are more resistant and less preferred for oviposition than susceptible North American species. We compared EAB oviposition preferences and bark and canopy volatile organic compound (VOC) emissions of resistant Manchurian ash and susceptible black ash, and examined relationships between VOC profiles and oviposition. In the field, black ash was highly preferred for oviposition while no eggs were laid on Manchurian ash, and we found clear differences in the VOC profiles of Manchurian and black ash. We detected 78 compounds emitted from these species, including 16 compounds that elicited EAB antennal activity in prior studies. Four compounds were unique to black and 11 to Manchurian ash. Emission rates of 14 canopy and 19 bark volatiles varied among the two species, including four previously reported as antennally active. Specifically, 7-epi-sesquithujene (bark) emissions were greater from black ash, while β-caryophyllene (canopy), linalool (bark), and α-cubebene (bark) were emitted at higher rates by Manchurian ash. No relationships were found between the emission rate of any single compound or group of compounds (e.g. monoterpenes) suggesting that preference may be based on complex profile combinations. This is the first study to directly compare VOCs of black and Manchurian ash as well as the first to examine bark- and canopy-specific VOCs. The unique bark and canopy VOC profiles of these two species implicates potentially important variation in VOCs between a closely related resistant and susceptible species that provides a foundation for future studies of host preferences of EAB.

Emerald ash borer (EAB) is an invasive beetle native to Asia that infests and kills ash (Fraxinus spp.) in North America. Previous experiments indicated that larvae feeding on co-evolved, resistant Manchurian ash (F. mandshurica) have increased antioxidant and quinone-protective enzyme activities compared to larvae feeding on susceptible North American species. Here, we examined mechanisms of host-generated oxidative and quinone-based stress and other putative defenses in Manchurian ash and the closely related and chemically similar, but susceptible, black ash (F. nigra), with and without exogenous application of methyl jasmonate (MeJA) to induce resistance mechanisms. Peroxidase activities were 4.6-13.3 times higher in Manchurian than black ash, although both species appeared to express the same three peroxidase isozymes. Additionally, peroxidase-mediated protein cross-linking activity was stronger in Manchurian ash. Polyphenol oxidase, β-glucosidase, chitinase, and lipoxygenase activities also were greater in Manchurian ash, but only lipoxygenase activity increased with MeJA application. Phloem H 2 O 2 levels were similar and were increased by MeJA application in both species. Lastly, trypsin inhibitor activity was detected in methanol and water extracts that were not allowed to oxidize, indicating the presence of phenolic-based trypsin inhibitors. However, no proteinaceous trypsin inhibitor activity was detected in either species. In response to MeJA application, Manchurian ash had higher trypsin inhibitor activity than black ash using the unoxidized water extracts, but no treatment effects were detected using methanol extracts. Based on these results we hypothesize that peroxidases, lignin polymerization, and quinone generation contribute to the greater resistance to EAB displayed by Manchurian ash.

Emerald ash borer (Agrilus planipennis Fairmaire), an invasive wood-boring beetle native to Asia, has killed hundreds of millions of ash trees since its accidental introduction into North America, resulting in widespread formation of canopy gaps and accumulations of coarse woody debris (CWD) in forests. The objective was to quantify effects of canopy gaps and CWD caused by early stages of emerald ash borer-induced ash mortality, and their interaction on ground beetle assemblages. The impact of canopy gaps and CWD varied, as gaps affected beetle assemblages in 2011, while effects of CWD were only observed in 2012. Gaps decreased beetle activity-abundance, and marginally decreased richness, driving changes in species composition, but evenness and diversity were unaffected. Effects of the CWD treatment alone were minimal, but CWD interacted with the canopy treatment, resulting in an increase in activity-abundance of ground beetles in canopy gaps without CWD, and a marginal increase in species richness in canopy gaps with CWD. Although there were some initial changes in species composition, these were ephemeral, suggesting that ground beetle assemblages may be resilient to disturbance caused by emerald ash borer. This study contributes to our understanding of the cascading ecological impacts of biological invasions on forest ecosystems., (© The Authors 2016. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Eradication is the deliberate elimination of a species from an area. Given that international quarantine measures can never be 100% effective, surveillance for newly arrived populations of nonnative species coupled with their eradication represents an important strategy for excluding potentially damaging insect species. Historically, eradication efforts have not always been successful and have sometimes been met with public opposition. But new developments in our understanding of the dynamics of low-density populations, the availability of highly effective treatment tactics, and bioeconomic analyses of eradication strategies offer new opportunities for developing more effective surveillance and eradication programs. A key component that connects these new developments is the harnessing of Allee effects, which naturally promote localized species extinction. Here we review these developments and suggest how research might enhance eradication strategies.

We review the literature on host resistance of ash to emerald ash borer (EAB, Agrilus planipennis), an invasive species that causes widespread mortality of ash. Manchurian ash (Fraxinus mandshurica), which coevolved with EAB, is more resistant than evolutionarily naïve North American and European congeners. Manchurian ash was less preferred for adult feeding and oviposition than susceptible hosts, more resistant to larval feeding, had higher constitutive concentrations of bark lignans, coumarins, proline, tyramine and defensive proteins, and was characterized by faster oxidation of phenolics. Consistent with EAB being a secondary colonizer of coevolved hosts, drought stress decreased the resistance of Manchurian ash, but had no effect on constitutive bark phenolics, suggesting that they do not contribute to increased susceptibility in response to drought stress. The induced resistance of North American species to EAB in response to the exogenous application of methyl jasmonate was associated with increased bark concentrations of verbascoside, lignin and/or trypsin inhibitors, which decreased larval survival and/or growth in bioassays. This finding suggests that these inherently susceptible species possess latent defenses that are not induced naturally by larval colonization, perhaps because they fail to recognize larval cues or respond quickly enough. Finally, we propose future research directions that would address some critical knowledge gaps., (© 2015 The Authors. New Phytologist © 2015 New Phytologist Trust.)

Protection of green ash trees (Fraxinus pennsylvanica Marshall) from the emerald ash borer (EAB), Agrilus planipennis Fairmaire, by soil applications of neonicotinoids (imidacloprid, clothianidin, and dinotefuran) was tested at five locations between 2005 and 2013. Application rate and spring versus fall application dates were evaluated in tests with neighborhood street trees and in one plantation of 65 ash trees. Insecticide treatments of ash trees at all five sites were initiated as the leading edge of the EAB invasion began to kill the first ash trees at each location. Trees were treated and evaluated at each site for 4 to 7 yr. Spring applications of imidacloprid were more efficacious than fall applications. Application rates of 0.8 g a.i./cm dbh or greater per year gave a higher level of protection and were more consistent than rates of 0.56 g a.i./cm dbh per year or less. The number of years between the first observation of canopy loss due to EAB and death of most of the control trees varied from three to seven years among test sites, depending on how many non-treated ash trees were nearby., (© The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Emerald ash borer, Agrilus planipennis Fairmaire, an Asian wood-boring beetle, has devastated ash (Fraxinus spp.) trees in North American forests and landscapes since its discovery there in 2002. In this study, we collected living larvae from EAB-resistant Manchurian ash (Fraxinus mandschurica), and susceptible white (Fraxinus americana) and green (Fraxinus pennsylvanica) ash hosts, and quantified the activity and production of selected detoxification, digestive, and antioxidant enzymes. We hypothesized that differences in larval physiology could be used to infer resistance mechanisms of ash. We found no differences in cytochrome P450, glutathione-S-transferase, carboxylesterase, sulfotransferase, and tryptic BApNAase activities between larvae feeding on different hosts. Despite this, Manchurian ash-fed larvae produced a single isozyme of low electrophoretic mobility that was not produced in white or green ash-fed larvae. Additionally, larvae feeding on white and green ash produced two serine protease isozymes of high electrophoretic mobility that were not observed in Manchurian ash-fed larvae. We also found lower activity of β-glucosidase and higher activities of monoamine oxidase, ortho-quinone reductase, catalase, superoxide dismutase, and glutathione reductase in Manchurian ash-fed larvae compared to larvae that had fed on susceptible ash. A single isozyme was detected for both catalase and superoxide dismutase in all larval groups. The activities of the quinone-protective and antioxidant enzymes are consistent with the resistance phenotype of the host species, with the highest activities measured in larvae feeding on resistant Manchurian ash. We conclude that larvae feeding on Manchurian ash could be under quinone and oxidative stress, suggesting these may be potential mechanisms of resistance of Manchurian ash to EAB larvae, and that quinone-protective and antioxidant enzymes are important counter-adaptations of larvae for dealing with these resistance mechanisms., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

The emerald ash borer (EAB; Agrilus planipennis Fairmaire) is causing widespread mortality of ash (Fraxinus spp.) in North America. To date, no mechanisms of host resistance have been identified against this pest. Methyl jasmonate was applied to susceptible North American and resistant Asian ash species to determine if it can elicit induced responses in bark that enhance resistance to EAB. In particular, phenolic compounds, lignin, and defense-related proteins were quantified, and compounds associated with resistance were subsequently tested directly against EAB larvae in bioassays with artificial diet. MeJA application decreased adult emergence in susceptible ash species, comparable to levels achieved by insecticide application. Concentration of the phenolic compound verbascoside sharply increased after MeJA application to green and white ash. When incorporated in an artificial diet, verbascoside decreased survival and growth of EAB neonates in a dose-dependent fashion. Lignin and trypsin inhibitors were also induced by MeJA, and analogs of both compounds reduced growth of EAB larvae in artificial diets. We conclude that the application of MeJA prior to EAB attack has the ability to enhance resistance of susceptible ash trees by inducing endogenous plant defenses, and report evidence that induction of verbascoside is a mechanism of resistance to EAB.

Exotic herbivorous insects have drastically and irreversibly altered forest structure and composition of North American forests. For example, emerald ash borer (Agrilus planipennis Fairmaire) from Asia has caused wide-scale mortality of ash trees (Fraxinus spp.) in eastern United States and Canada. We studied the effects of forest changes resulting from emerald ash borer invasion on epigaeic or ground beetles (Coleoptera: Carabidae) along a gradient of ash dieback and gap sizes in southeastern Michigan. Ground beetles were sampled in hydric, mesic, and xeric habitats in which black (Fraxinus nigra Marshall), green (Fraxinus pennsylvanica Marshall), and white (Fraxinus americana L.) ash were the most common species, respectively. During 2006-2007, we trapped 2,545 adult ground beetles comprising 52 species. There was a negative correlation between percent ash tree mortality in 2006 and catches of all beetles. Catches of Agonum melanarium Dejean (in 2006) and Pterostichus mutus (Say) (in 2006-2007) were negatively correlated with tree mortality and gap size, respectively. However, catches of Pterostichus corvinus Dejean were positively correlated with gap size in 2006. As ash mortality and average gap size increased from 2006 to 2007, catches of all beetles as well as P. mutus and Pterostichus stygicus (Say) increased (1.3-3.9 times), while species diversity decreased, especially in mesic and xeric stands. Cluster analysis revealed that beetle assemblages in hydric and mesic stand diverged (25 and 40%, respectively) in their composition from 2006 to 2007, and that hydric stands had the most unique beetle assemblages. Overall, epigaeic beetle assemblages were altered in ash stands impacted by emerald ash borer; however, these impacts may dissipate as canopy gaps close.

The invasive emerald ash borer (EAB) beetle is a significant threat to the survival of North American ash. In previous work, we identified putative biochemical and molecular markers of constitutive EAB resistance in Manchurian ash, an Asian species co-evolved with EAB. Here, we employed high-throughput high-performance liquid chromatography with photodiode array detection and mass spectrometry (HPLC-PDA-MS) to characterize the induced response of soluble phloem phenolics to EAB attack in resistant Manchurian and susceptible black ash under conditions of either normal or low water availability, and the effects of water availability on larval performance. Total larval mass per tree was lower in Manchurian than in black ash. Low water increased larval numbers and mean larval mass overall, but more so in Manchurian ash. Low water did not affect levels of phenolics in either host species, but six phenolics decreased in response to EAB. In both ashes, pinoresinol A was induced by EAB, especially in Manchurian ash. Pinoresinol A and pinoresinol B were negatively correlated with each other in both species. The higher accumulation of pinoresinol A in Manchurian ash after attack may help explain the resistance of this species to EAB, but none of the responses measured here could explain increased larval performance in trees subjected to low water availability., (© 2013 John Wiley & Sons Ltd.)

Since its accidental introduction from Asia, emerald ash borer (EAB), Agrilus planipennis Fairmaire (Coleoptera: Buprestidae), has killed millions of ash trees in North America. As it continues to spread, it could functionally extirpate ash with devastating economic and ecological impacts. Little was known about EAB when it was first discovered in North America in 2002, but substantial advances in understanding of EAB biology, ecology, and management have occurred since. Ash species indigenous to China are generally resistant to EAB and may eventually provide resistance genes for introgression into North American species. EAB is characterized by stratified dispersal resulting from natural and human-assisted spread, and substantial effort has been devoted to the development of survey methods. Early eradication efforts were abandoned largely because of the difficulty of detecting and delineating infestations. Current management is focused on biological control, insecticide protection of high-value trees, and integrated efforts to slow ash mortality.

Bronze birch borer (Agrilus anxius Gory) (Coleoptera: Buprestidae), a specialist wood-borer endemic to North America, is prone to periodic outbreaks that have caused widespread mortality of birch (Betula spp.) in boreal and north temperate forests. It is also the key pest of birch in ornamental landscapes. Amenity plantings have extended the distribution of birch in North America, for which we report an updated map. Life history and phenology also are summarized. Larvae feed primarily on phloem tissue of stems and branches, which can girdle and kill trees. Stressors such as drought, elevated temperature, and defoliation predispose trees to bronze birch borer colonization and trigger outbreaks, which implicates the availability of suitable host material in the bottom-up regulation of populations. Stress imposed by climate change may increase the frequency of outbreaks and alter the distribution of birch. Bronze birch borer has a diverse array of natural enemies, but their role in top-down population regulation has not been studied. There is substantial interspecific variation in resistance to this insect. North American species share a coevolutionary history with bronze birch borer and are much more resistant than Eurasian species, which are evolutionarily naïve. Potential resistance mechanisms are reviewed. The high susceptibility of Eurasian birch species and climatic similarities of North America and Eurasia create high risk of widespread birch mortality in Eurasia if the borer was inadvertently introduced. Bronze birch borer can be managed in amenity plantings through selection of resistant birch species, plant health care practices, and insecticides.

The emerald ash borer (Agrilus planipennis, EAB) is an invasive wood-borer indigenous to Asia and is responsible for widespread ash (Fraxinus spp.) mortality in the U.S. and Canada. Resistance and susceptibility to EAB varies among Fraxinus spp., which is a result of their co-evolutionary history with the pest. We characterized constitutive phenolic profiles and lignin levels in the phloem of green, white, black, blue, European, and Manchurian ash. Phloem was sampled twice during the growing season, coinciding with phenology of early and late instar EAB. We identified 66 metabolites that displayed a pattern of variation, which corresponded strongly with phylogeny. Previously identified lignans and lignan derivatives were confirmed to be unique to Manchurian ash, and may contribute to its high level of resistance to EAB. Other compounds that had been considered unique to Manchurian ash, including hydroxycoumarins and the phenylethanoids calceolarioside A and B, were detected in closely related, but susceptible species, and thus are unlikely to contribute to EAB resistance of Manchurian ash. The distinct phenolic profile of blue ash may contribute to its relatively high resistance to EAB.

Invasive species increasingly threaten ecosystems, food production, and human welfare worldwide. Hundreds of eradication programs have targeted a wide range of nonnative insect species to mitigate the economic and ecological impacts of biological invasions. Many such programs used multiple tactics to achieve this goal, but interactions between tactics have received little formal consideration, specifically as they interact with Allee dynamics. If a population can be driven below an Allee threshold, extinction becomes more probable because of factors such as the failure to find mates, satiate natural enemies, or successfully exploit food resources, as well as demographic and environmental stochasticity. A key implication of an Allee threshold is that the population can be eradicated without the need and expense of killing the last individuals. Some combinations of control tactics could interact with Allee dynamics to increase the probability of successful eradication. Combinations of tactics can be considered to have synergistic (greater efficiency in achieving extinction from the combination), additive (no improvement over single tactics alone), or antagonistic (reduced efficiency from the combination) effects on Allee dynamics. We highlight examples of combinations of tactics likely to act synergistically, additively, or antagonistically on pest populations. By exploiting the interacting effects of multiple tactics on Allee dynamics, the success and cost-effectiveness of eradication programs can be enhanced.

Outbreaks of bronze birch borer (BBB) (Agrilus anxius), a wood-boring beetle endemic to North America, have been associated with widespread mortality of birch (Betula spp.). There is substantial inter- and intra-specific variation in birch resistance to BBB. Species endemic to North America, such as paper birch (B. papyrifera), have coevolved with BBB and are more resistant than European and Asian birch species, such as European white birch (B. pendula), which lack an evolutionary history with BBB. Borer larvae feed on stem phloem tissue. Therefore, in search of potential resistance mechanisms against BBB, we compared the constitutive phenolic profile of stem phloem tissue of paper birch with that of European white birch. We also analyzed intraspecific variation in phenolic composition among clones and/or half-siblings of both species. Three phenolics (coumaroylquinic acid, betuloside pentoside A, and a diarylheptanoid hexoside) were detected only in paper birch, and concentrations of six other phenolics were significantly higher in paper birch. These differences may contribute to the high resistance of paper birch to BBB relative to European white birch. There was significant intraspecific variation in four of 17 phenolics found in paper birch and in five of 14 found in European white birch, but clones and half-siblings within each species could not be distinguished by phenolic composition using multivariate analysis.

Bronze birch borer (Agrilus anxius Gory) is the key pest of birches (Betula spp.) in North America, several of which have been recommended for ornamental landscapes based on anecdotal reports of borer resistance that had not been confirmed experimentally. In a 20-yr common garden experiment initiated in 1979 in Ohio, North American birch species, including paper birch (Betula papyrifera Marshall), 'Whitespire' gray birch (Betula populifolia Marshall), and river birch (Betula nigra L.), were much more resistant to bronze birch borer than species indigenous to Europe and Asia, including European white birch (Betula pendula Roth), downy birch (Betula pubescens Ehrh.), monarch birch (Betula maximowicziana Regel), and Szechuan white birch (Betula szechuanica Jansson). Within 8 yr of planting, every European white, downy, and Szechuan birch had been colonized and killed, although 100% of monarch birch had been colonized and 88% of these plants were killed after nine years. Conversely, 97% of river birch, 76% of paper birch, and 73% Whitespire gray birch were alive 20 yr after planting, and river birch showed no evidence of colonization. This pattern is consistent with biogeographic theory of plant defense: North American birch species that share a coevolutionary history with bronze birch borer were much more resistant than naïve hosts endemic to Europe and Asia, possibly by virtue of evolution of targeted defenses. This information suggests that if bronze birch borer were introduced to Europe or Asia, it could threaten its hosts there on a continental scale. This study also exposed limitations of anecdotal observation as evidence of host plant resistance.

Phytophagous insects frequently encounter reactive oxygen species (ROS) from exogenous and endogenous sources. To overcome the effect of ROS, insects have evolved a suite of antioxidant defense genes such as superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) and glutathione peroxidase (GPX). The emerald ash borer (Agrilus planipennis Fairmaire), an exotic invasive insect pest from Asia has killed millions of ash trees and continues to invade North America at a rapid pace. From an on-going expressed sequence tag (EST) project of A. planipennis larval tissues, we identified ESTs coding for a Cu-Zn SOD (ApSOD1), a CAT (ApCAT1) and a GPX (ApGPX1). A multiple sequence alignment of the derived A. planipennis sequences revealed high homology with other insect sequences at the amino acid level. Phylogenetic analysis of ApSOD1 grouped it with Cu-Zn SODs of other insect taxa. Quantitative real time PCR (qRT-PCR) analysis in different larval tissues (midgut, fat body, Malpighian tubule and cuticle) revealed high mRNA levels of ApCAT1 in the midgut. Interestingly, high mRNA levels for both ApSOD1 and ApGPX1 were observed in the Malpighian tubules. Assay of mRNA levels in developmental stages (larva, prepupa and adults) by qRT-PCR indicated high transcript levels of ApCAT1 and ApGPX1 in larval and prepupal stages with a decline in adults. On the other hand, the transcript levels of ApSOD1 were observed to be constitutive in all the developmental stages assayed. Results obtained reflect a plausible role of these A. planipennis antioxidant genes in quenching ROS from both diet (ash allelochemicals) as well as endogenous sources. These studies further help in understanding the adaptation/invasiveness of A. planipennis., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

We examined the extent to which three Fraxinus cultivars and a wild population that vary in their resistance to Emerald Ash Borer (EAB) could be differentiated on the basis of a suite of constitutive chemical defense traits in phloem extracts. The EAB-resistant Manchurian ash (F. mandshurica, cv. Mancana) was characterized by having a rapid rate of wound browning, a high soluble protein concentration, low trypsin inhibitor activities, and intermediate levels of peroxidase activity and total soluble phenolic concentration. The EAB-susceptible white ash (F. americana, cv. Autumn Purple) was characterized by a slow wound browning rate and low levels of peroxidase activity and total soluble phenolic concentrations. An EAB-susceptible green ash cultivar (F. pennsylvanica, cv. Patmore) and a wild accession were similar to each other on the basis of several chemical defense traits, and were characterized by high activities of peroxidase and trypsin inhibitor, a high total soluble phenolic concentration, and an intermediate rate of wound browning. Lignin concentration and polyphenol oxidase activities did not differentiate resistant and susceptible species. Of 33 phenolic compounds separated by HPLC and meeting a minimum criterion for analysis, nine were unique to Manchurian ash, five were shared among all species, and four were found in North American ashes and not in the Manchurian ash. Principal components analysis revealed clear separations between Manchurian, white, and green ashes on the basis of all phenolics, as well as clear separations on the basis of quantities of phenolics that all species shared. Variation in some of these constitutive chemical defense traits may contribute to variation in resistance to EAB in these species.

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.

Background: Ash (Fraxinus spp.) is a dominant tree species throughout urban and forested landscapes of North America (NA). The rapid invasion of NA by emerald ash borer (Agrilus planipennis), a wood-boring beetle endemic to Eastern Asia, has resulted in the death of millions of ash trees and threatens billions more. Larvae feed primarily on phloem tissue, which girdles and kills the tree. While NA ash species including black (F. nigra), green (F. pennsylvannica) and white (F. americana) are highly susceptible, the Asian species Manchurian ash (F. mandshurica) is resistant to A. planipennis perhaps due to their co-evolutionary history. Little is known about the molecular genetics of ash. Hence, we undertook a functional genomics approach to identify the repertoire of genes expressed in ash phloem., Methodology and Principal Findings: Using 454 pyrosequencing we obtained 58,673 high quality ash sequences from pooled phloem samples of green, white, black, blue and Manchurian ash. Intriguingly, 45% of the deduced proteins were not significantly similar to any sequences in the GenBank non-redundant database. KEGG analysis of the ash sequences revealed a high occurrence of defense related genes. Expression analysis of early regulators potentially involved in plant defense (i.e. transcription factors, calcium dependent protein kinases and a lipoxygenase 3) revealed higher mRNA levels in resistant ash compared to susceptible ash species. Lastly, we predicted a total of 1,272 single nucleotide polymorphisms and 980 microsatellite loci, among which seven microsatellite loci showed polymorphism between different ash species., Conclusions and Significance: The current transcriptomic data provide an invaluable resource for understanding the genetic make-up of ash phloem, the target tissue of A. planipennis. These data along with future functional studies could lead to the identification/characterization of defense genes involved in resistance of ash to A. planipennis, and in future ash breeding programs for marker development.

The emerald ash borer (Agrilus planipennis) is an invasive wood-boring beetle that has killed millions of ash trees since its accidental introduction to North America. All North American ash species (Fraxinus spp.) that emerald ash borer has encountered so far are susceptible, while an Asian species, Manchurian ash (F. mandshurica), which shares an evolutionary history with emerald ash borer, is resistant. Phylogenetic evidence places North American black ash (F. nigra) and Manchurian ash in the same clade and section, yet black ash is highly susceptible to the emerald ash borer. This contrast provides an opportunity to compare the genetic traits of the two species and identify those with a potential role in defense/resistance. We used Difference Gel Electrophoresis (DIGE) to compare the phloem proteomes of resistant Manchurian to susceptible black, green, and white ash. Differentially expressed proteins associated with the resistant Manchurian ash when compared to the susceptible ash species were identified using nano-LC-MS/MS and putative identities assigned. Proteomic differences were strongly associated with the phylogenetic relationships among the four species. Proteins identified in Manchurian ash potentially associated with its resistance to emerald ash borer include a PR-10 protein, an aspartic protease, a phenylcoumaran benzylic ether reductase (PCBER), and a thylakoid-bound ascorbate peroxidase. Discovery of resistance-related proteins in Asian species will inform approaches in which resistance genes can be introgressed into North American ash species. The generation of resistant North American ash genotypes can be used in forest ecosystem restoration and urban plantings following the wake of the emerald ash borer invasion.

Background: The insect midgut and fat body represent major tissue interfaces that deal with several important physiological functions including digestion, detoxification and immune response. The emerald ash borer (Agrilus planipennis), is an exotic invasive insect pest that has killed millions of ash trees (Fraxinus spp.) primarily in the Midwestern United States and Ontario, Canada. However, despite its high impact status little knowledge exists for A. planipennis at the molecular level., Methodology and Principal Findings: Newer-generation Roche-454 pyrosequencing was used to obtain 126,185 reads for the midgut and 240,848 reads for the fat body, which were assembled into 25,173 and 37,661 high quality expressed sequence tags (ESTs) for the midgut and the fat body of A. planipennis larvae, respectively. Among these ESTs, 36% of the midgut and 38% of the fat body sequences showed similarity to proteins in the GenBank nr database. A high number of the midgut sequences contained chitin-binding peritrophin (248)and trypsin (98) domains; while the fat body sequences showed high occurrence of cytochrome P450s (85) and protein kinase (123) domains. Further, the midgut transcriptome of A. planipennis revealed putative microbial transcripts encoding for cell-wall degrading enzymes such as polygalacturonases and endoglucanases. A significant number of SNPs (137 in midgut and 347 in fat body) and microsatellite loci (317 in midgut and 571 in fat body) were predicted in the A. planipennis transcripts. An initial assessment of cytochrome P450s belonging to various CYP clades revealed distinct expression patterns at the tissue level., Conclusions and Significance: To our knowledge this study is one of the first to illuminate tissue-specific gene expression in an invasive insect of high ecological and economic consequence. These findings will lay the foundation for future gene expression and functional studies in A. planipennis.

In 2007, we surveyed the alien and endemic scolytine (bark and ambrosia beetles) fauna of northeastern Ohio, and for the most abundant species, we characterized their seasonal activity and response to three semiochemical baits. In total ,5,339 scolytine beetles represented by 47 species and 29 genera were caught in Lindgren funnel traps. Three species constituted 57% of the total catch, including Xylosandrus germanus (Blandford), Tomicus piniperda (L.), and Dryocoetes autographus (Ratzeburg). Of the total captured, 32% of the species and approximately 60% of the individuals were exotic, suggesting that exotic species numerically dominate the scolytine fauna in some urban areas. More native and exotic species were caught in traps baited with ethanol alone than in traps baited with other lures. However, significantly more individuals, especially of T. piniperda, D. autographus, Gnathotrichus materiarius (Fitch), and Ips grandicollis (Eichhoff), and species were caught in traps baited with ethanol plus alpha-pinene than in traps baited with ethanol alone or the exotic Ips lure. This suggests that among these baits, the ethanol plus alpha-pinene baits may be useful in maximizing scolytine beetle catches of these species within this region. Species diversity and richness for both native and exotic beetles was greatest in traps baited with ethanol alone. The period of peak trap capture varied depending upon species: X. germanus was most abundant in traps in mid-May and early-August; T. piniperda in mid-May; D. autographus in early June, mid-July, and mid-September; Anisandrus sayi Hopkins and G. materiarius in mid-May, mid-July, and early September; and I. grandicollis in early April, mid-July, and late September.

Paper birch (Betula papyrifera Marsh) seedlings were grown in a greenhouse in either subsoil or topsoil in factorial combination with two fertilization and drought regimes to investigate how different soil environments and nutrient availability drive belowground partitioning between growth, secondary metabolism and ectomycorrhizal (EM) associations, and impact drought tolerance of paper birch. Root and total seedling dry biomass, starch, soluble sugars, soluble phenolics, lignin and EM abundance were quantified. In unfertilized topsoil, total plant biomass and root biomass were approximately nine times higher than in unfertilized subsoil, but the root weight ratios did not differ between soils. Root soluble phenolics and lignin were higher in unfertilized subsoil than in unfertilized topsoil, whereas EM abundance was significantly higher in unfertilized topsoil than in unfertilized subsoil. In topsoil, fertilization decreased root biomass and EM abundance and increased root phenolics and lignin. In contrast, fertilization of subsoil increased root biomass but decreased root phenolics and lignin, while EM abundance was unaffected. In both soil types, fertilization reduced root weight ratios. Across soil types, EM abundance was negatively correlated with root soluble sugars, root phenolics and lignin, but this was driven mainly by the responses in the topsoil treatment. Our results show that soil fertility mediates carbon tradeoffs among defense, growth and EM associations.

Although current USDA-APHIS standards suggest that a core temperature of 71.1 degrees C (160 degrees F) for 75 min is needed to adequately sanitize emerald ash borer, Agrilus planipennis Fairmaire-infested firewood, it is unclear whether more moderate (and economical) treatment regimes will adequately eradicate emerald ash borer larvae and prepupae from ash firewood. We constructed a small dry kiln in an effort to emulate the type of technology a small- to medium-sized firewood producer might use to examine whether treatments with lower temperature and time regimes successfully eliminate emerald ash borer from both spilt and roundwood firewood. Using white ash (Fraxinus americana L.) firewood collected from a stand with a heavy infestation of emerald ash borer in Delaware, OH, we treated the firewood using the following temperature and time regime: 46 degrees C (114.8 degrees F) for 30 min, 46 degrees C (114.8 degrees F) for 60 min, 56 degrees C (132.8 degrees F) for 30 min, and 56 degrees C (132.8 degrees F) for 60 min. Temperatures were recorded for the outer 2.54-cm (1-in.) of firewood. After treatment, all firewood was placed under mesh netting and emerald ash borer were allowed to develop and emerge under natural conditions. No treatments seemed to be successful at eliminating emerald ash borer larvae and perpupae as all treatments (including two nontreated controls) experienced some emerald ash borer emergence. However, the 56 degrees C (132.8 degrees F) treatments did result in considerably less emerald ash borer emergence than the 46 degrees C (114.8 degrees F) treatments. Further investigation is needed to determine whether longer exposure to the higher temperature (56 degrees C) will successfully sanitize emerald ash borer-infested firewood.

Urbanization affects communities of herbivorous arthropods and provides opportunities for dramatic changes in their abundance and richness. Underlying these changes are creation of impervious surfaces; variation in the density, diversity, and complexity of vegetation; and maintenance practices including pulsed inputs of fertilizers, water, and pesticides. A rich body of knowledge provides theoretical underpinnings for predicting and understanding impacts of urbanization on arthropods. However, relatively few studies have elucidated mechanisms that explain patterns of insect and mite abundance and diversity across urbanization gradients. Published accounts suggest that responses to urbanization are often taxon specific, highly variable, and linked to properties of urbanization that weaken top-down and/or bottom-up processes, thereby destabilizing populations of herbivores and their natural enemies. In addition to revealing patterns in diversity and abundance of herbivores across urbanization gradients, a primary objective of this review is to examine mechanisms underlying these patterns and to identify potential hypotheses for future testing.

Despite their economic and ecological importance, defense responses of conifers to pests are little understood. In a 3-year experiment, we monitored systemic fungal (Diplodia pinea)- and insect (Neodiprion sertifer)-induced defense protein activities and total soluble proteins in needles and phloem of Austrian pine (Pinus nigra) across a soil fertility gradient. In both years, total soluble protein content of foliage and phloem declined with increasing fertility across induction treatments, while defensive protein activities generally increased with increasing fertility. In 2005, total soluble protein content in branch phloem was increased by fungal inoculation of the stem. Peroxidase activity was suppressed in needles by insect defoliation in 2006, while polyphenol oxidase activity was systemically induced in branch phloem by insect attack in 2005. Trypsin inhibitor activities in phloem did not respond to any induction or fertility treatment. Nutritive quality of Austrian pine tissue declined with increasing fertility, while several protein-based defenses simultaneously increased.

We conducted a 3-yr study to compare the susceptibility of selected North American ash and an Asian ash species to emerald ash borer, Agrilus planipennis Fairmaire, an invasive wood-boring beetle introduced to North America from Asia. Because of a coevolutionary relationship between Asian ashes and emerald ash borer, we hypothesized an Asian ash species, Manchurian ash, is more resistant to the beetle than its North American congeners. Consistent with our hypothesis, Manchurian ash experienced far less mortality and yielded far fewer adult beetles than several cultivars of North American green and white ash. Surprisingly, a black ash (North American) x Manchurian ash hybrid was highly susceptible to emerald ash borer, indicating this cultivar did not inherit emerald ash borer resistance from its Asian parent. A corollary study investigated the efficacy of soil-applied imidacloprid, a systemic, neonicotinoid insecticide, for controlling emerald ash borer in each of the five cultivars. Imidacloprid had no effect on emerald ash borer colonization of Manchurian ash, which was low in untreated and treated trees. In contrast, imidacloprid did enhance survival of the North American and hybrid cultivars and significantly reduced the number of emerald ash borer adults emerging from green and white ash cultivars. We identify a possible mechanism of resistance of Manchurian ash to emerald ash borer, which may prove useful for screening, selecting, and breeding emerald ash borer-resistant ash trees.

The mechanisms and conditions affecting expression of systemic induced resistance (SIR) in pine are not clearly understood. Two hypotheses were tested here: that SIR against a pathogen induced by either a pathogen or an insect involves coordinated shifts in phloem secondary metabolism; and that fertility affects the production of these compounds. To test these hypotheses, a tripartite system was used comprising Austrian pine (Pinus nigra) grown under three different fertility regimes, the fungal pathogen Diplodia pinea, and the defoliator Neodiprion sertifer. Fungal induction led to systemic accumulation of lignin, phenolic glycosides and stilbenes, whereas insect defoliation led to an increase in germacrene D concentration in branch phloem. Fertility affected the concentrations of only the phenolic glycosides. Multivariate analyses showed coregulation of compounds within at least three consistent groupings: phenolic glycosides, stilbenes and monoterpenes. As groups and as individual compounds, accumulation of phenolic glycosides and stilbenes was negatively correlated with disease susceptibility. The experimental manipulation of the phenolics and terpenoids metabolic networks achieved in this study by biotic induction and changes in nutrient availability suggests that lignin, phenolic glycosides and stilbenes are important biochemical factors in the expression of SIR against the pathogen in this system.

Recent studies have investigated interspecific variation in resistance of ash (Fraxinus spp.) to the exotic wood-boring beetle, emerald ash borer (EAB, Agrilus planipennis). Manchurian ash (Fraxinus mandshurica) is an Asian species that has coevolved with EAB. It experiences little EAB-induced mortality compared to North American ashes. Host phloem chemistry, both constitutive and induced, might partly explain this interspecific variation in resistance. We analyzed the constitutive phloem chemistry of three ash species: Manchurian ash and North American white (Fraxinus americana) and green (Fraxinus pennsylvanica) ash. Analysis of the crude phloem extracts revealed the presence of an array of phenolic compounds including hydroxycoumarins, a monolignol, lignans, phenylethanoids, and secoiridoids. Both qualitative and quantitative differences were observed among the three ash species. Hydroxycoumarins and the phenylethanoids, calceloariosides A and B, were present only in the phloem of Manchurian ash and might represent a mechanism of resistance against EAB.

Here, the growth-differentiation balance hypothesis (GDBH) was tested by quantifying temporal variation in the relative growth rate (RGR), net assimilation rate (NAR), and phenylpropanoid concentrations of two willow species (Salix sericea and Salix eriocephala) across five fertility levels. Initially, RGR increased and total phenylpropanoids declined (although every individual phenolic did not) as fertility increased, but NAR was unaffected. Subsequently, NAR and phenylpropanoids declined in the low fertility treatment, generating a quadratic response of secondary metabolism across the nutrient gradient. As above- and below-ground growth rates equilibrated, NAR and phenylpropanoids increased in the low fertility treatment, re-establishing a negative linear effect of fertility on secondary metabolism. A transient quadratic response of secondary metabolism is predicted when GDBH is integrated with models of optimal phenotypic plasticity, occurring when low NAR imposes carbon constraints on secondary metabolism in low nutrient environments. Once plants acclimate to nutrient limitation, the equilibrium allocation state is predicted to be a negative correlation between growth and secondary metabolism. Although both willow species generally responded according to GDBH, the complexity observed suggests that prediction of the effects of nutrient availability on secondary metabolism (and other plastic responses) in specific cases requires a priori knowledge of the physiological status of the plant and soil nutrient availability.

The growth-differentiation balance hypothesis (GDBH) postulates that variation in resource availability can increase or decrease allocation to secondary metabolism, depending on how growth is affected relative to carbon assimilation. Growth and leaf area of black poplar (Populus nigra) increased substantially in response to increased nutrient availability, while net assimilation rate and photosynthesis were less strongly affected. In response, total phenolic glycoside concentrations declined, which is consistent with GDBH. Drought stress decreased net assimilation rate and photosynthesis as well as growth, while increasing total phenolic glycoside concentrations. This pattern does not follow GDBH, which predicts lower secondary metabolism when resource limitation decreases both growth and carbon assimilation. However, there was a strong negative correlation between growth and total phenolic glycoside concentration consistent with a trade-off between primary and secondary metabolism, a key premise of GDBH. Drought decreased the growth of gypsy moth (Lymantria dispar) larvae but had no effect on whitemarked tussock moth (Orgyia leucostigma). Increased nutrient availability had a positive linear effect on growth of whitemarked tussock moth, but no effect on gypsy moth. Treatment effects on gypsy moth corresponded closely with effects on total phenolic glycosides, whereas effects on whitemarked tussock moth more closely tracked changes in nutritional quality. Localized gypsy moth herbivory elicited rapid induced resistance to gypsy moth, with the effect being independent of water and nutrient availability, but did not affect whitemarked tussock moth, indicating that the effects of biotic and abiotic stress on insect resistance of trees can be species-specific.

Proanthocyanidins (condensed tannins) frequently need to be quantified in large numbers of samples in food, plant, and environmental studies. An automated colorimetric method to quantify proanthocyanidins with sulfuric acid (H(2)SO(4)) was therefore developed for use in a continuous flow analyzer. Assay conditions were optimized using 50% methanol extracts of paper birch, sugar maple, and quaking aspen leaves. Short extraction times and centrifugation of samples prevented proanthocyanidin degradation that otherwise occurred in 50% methanol extracts of aspen leaves. Extraction of birch and maple proanthocyanidins with 50% methanol was comparable to or better than that with 70% acetone. Proanthocyanidin levels in aspen were lower when extracted with aqueous methanol, but relative differences among samples were consistent with those found in aqueous acetone extracts. Results from the automated sulfuric acid assay were highly correlated with those of the conventional BuOH-HCl method for proanthocyanidins and, except for birch, with the Folin--Denis assay for total phenolics. This new technique significantly improves assay processing rate and repeatability compared to conventional colorimetric proanthocyanidin assays.

Leaves of transgenic sweetgum (Liquidambar styraciflua) trees that expressed tobacco anionic peroxidase were compared with leaves of L. styraciflua trees that did not express the tobacco enzyme. Leaves of the transgenic trees were generally more resistant to feeding by caterpillars and beetles than wild-type leaves. However, as for past studies with transgenic tobacco and tomato expressing the tobacco anionic peroxidase, the degree of relative resistance depended on the size of insect used and the maturity of the leaf. Decreased growth of gypsy moth larvae appeared mainly due to decreased consumption, and not changes in the nutritional quality of the foliage. Transgenic leaves were more susceptible to feeding by the corn earworm, Helicoverpa zea. Thus, it appears the tobacco anionic peroxidase can contribute to insect resistance, but its effects are more predictable when it is expressed in plant species more closely related to the original gene source.