Rapid induced resistance of silver birch affects both innate immunity and performance of gypsy moths: the role of plant chemical defenses

In this study we tested the effects of rapid induced resistance of the silver birch, Betula pendula, on the performance and immune defense of the gypsy moth, Lymantria dispar. We also measured the effects of defoliation on the concentrations of plant secondary metabolites, particularly on phenolics and terpenoids. It was found that severe natural defoliation (by moth larvae) of silver birch led to an increase in lipophilic flavonoids on the leaf surface. The concentration of some simple phenolics and monoterpenes (linalool and geraniol) also increased, while that of several glycosides of quercetin decreased. The female pupal weights and survival rates of moths decreased, and larval development time increased, when the insects fed on defoliated trees. However, the feeding of caterpillars with the leaves of defoliated trees led to an increase in lysozyme-like activity in their hemolymph, with an increase in their ability to encapsulate potential parasites. Our data show that the silver birch deploys a rapid chemical defense against gypsy moth larvae. We suggest that lipophilic flavonoids are important compounds in the direct silver birch defense against L. dispar caterpillars. The increased strength of immune defense of insects exposed to trees that had deployed a rapid induced resistance may be an adaptation of the herbivores to resist the rising density of parasites when host population density is high.     

Asynchrony between Host Plant and Insects-Defoliator within a Tritrophic System: The Role of Herbivore Innate Immunity

The effects of asynchrony in the phenology of spring-feeding insect-defoliators and their host plants on insects’ fitness, as well as the importance of this effect for the population dynamics of outbreaking species of insects, is a widespread and well-documented phenomenon. However, the spreading of this phenomenon through the food chain, and especially those mechanisms operating this spreading, are still unclear. In this paper, we study the effect of seasonally declined leafquality (estimated in terms of phenolics and nitrogen content) on herbivore fitness, immune parameters and resistance against pathogen by using the silver birch Betula pendula—gypsy moth Lymantria dispar—nucleopolyhedrovirus as the tritrophic system. We show that a phenological mismatch induced by the delay in the emergence of gypsy moth larvae and following feeding on mature leaves has negative effects on the female pupal weight, on the rate of larval development and on the activity of phenoloxidase in the plasma of haemolymph. In addition, the larval susceptibility to exogenous nucleopolyhydrovirus infection as well as covert virus activation were both enhanced due to the phenological mismatch. The observed effects of phenological mismatch on insect-baculovirus interaction may partially explain the strong and fast fluctuations in the population dynamics of the gypsy moth that is often observed in the studied part of the defoliator area. This study also reveals some indirect mechanisms of effect related to host plant quality, which operate through the insect innate immune status and affect resistance to both exogenous and endogenous virus.     

Cd胁迫对舞毒蛾幼虫食物利用的影响及其Cd的外泌机制

为探明植食性昆虫对受重金属胁迫的寄主植物的生理生态响应机制,本研究用Cd胁迫下银中杨的叶片饲养舞毒蛾幼虫,分析舞毒蛾幼虫对食物的利用情况以及其对Cd的排毒代谢机制.结果表明: 取食Cd胁迫下银中杨的叶片后,舞毒蛾3～6龄幼虫体内的Cd浓度和Cd含量均显著高于对照,但随着幼虫龄期增长,其体内Cd浓度显著降低,而Cd含量有不同程度的提高;舞毒蛾幼虫粪便和虫蜕中的Cd浓度均显著高于对照;舞毒蛾3～5龄幼虫的食物消耗率显著高于对照,而转化率显著低于对照;3～4龄幼虫的食物利用率均与对照差异不显著,但在5龄时显著低于对照.说明在Cd胁迫下,舞毒蛾幼虫能通过有效的排毒代谢途径将体内富集的部分Cd排出体外,且高龄幼虫的排毒代谢能力强于低龄幼虫;舞毒蛾幼虫体质量的增加会对体内的Cd浓度形成一种稀释效应;舞毒蛾幼虫能通过调整食物消耗率和转化率之间的比例,来维持其正常生长发育所需的食物利用率,但超过一定限度后仍会造成食物利用率降低.     

Decline in gypsy moth (Lymantria dispar) performance in an elevated CO2 atmosphere depends upon host plant species

Plant species differ broadly in their responses to an elevated CO₂ atmosphere, particularly in the extent of nitrogen dilution of leaf tissue. Insect herbivores are often limited by the availability of nutrients, such as nitrogen, in their host plant tissue and may therefore respond differentially on different plant species grown in CO₂-enriched environments. We reared gyspy moth larvae (Lymantria dispar) in situ on seedlings of yellow birch (Betula allegheniensis) and gray birch (B. populifolia) grown in an ambient (350 ppm) or elevated (700 ppm) CO₂ atmosphere to test whether larval responses in the elevated CO₂ atmosphere were species-dependent. We report that female gypsy moths (Lymantria dispar) reared on gray birch (Betula populifolia) achieved similar pupal masses on plants grown at an ambient or an elevated CO₂ concentration. However, on yellow birch (B. allegheniensis), female pupal mass was 38% smaller on plants in the elevated-CO₂ atmosphere. Larval mortality was significantly higher on yellow birch than gray birch, but did not differ between the CO₂ treatments. Relative growth rate declined more in the elevated CO₂ atmosphere for larvae on yellow birch than for those on gray birch. In preference tests, larvae preferred ambient over elevated CO₂-grown leaves of yellow birch, but showed no preference between gray birch leaves from the two CO₂ atmospheres. This differential response of gypsy moths to their host species corresponded to a greater decline in leaf nutritional quality in the elevated CO₂ atmosphere in yellow birch than in gray birch. Leaf nitrogen content of yellow birch dropped from 2.68% to 1.99% while that of gray birch leaves only declined from 3.23% to 2.63%. Meanwhile, leaf condensed tannin concentration increased from 8.92% to 11.45% in yellow birch leaves while gray birch leaves only increased from 10.72% to 12.34%. Thus the declines in larval performance in a future atmosphere may be substantial and host-species-specific.     

Influence of Diet and Density on Laboratory Cannibalism Behaviors in Gypsy Moth Larvae (Lymantria dispar L.)

The gypsy moth (Lymantria dispar) is an insect folivore that feeds on a broad range of hosts, and undergoes intermittent outbreaks that cause extensive tree mortality. Like many other herbivorous insects, gypsy moth larvae consume a substrate that is low in nitrogen. Gypsy moth larvae have been known to cannibalize under crowded conditions in the laboratory. In this study, we assessed the influence of nitrogen and density on cannibalism behavior in gypsy moth larvae. Cannibalism rates increased with decreased nitrogen and increased density. There was no interaction between these two parameters. Developmental experiments confirmed that low dietary nitrogen is detrimental, in agreement with previous studies. In a second experiment, we assessed the influence of previous cannibalism experiences on subsequent cannibalism behavior. Gypsy moth larvae that had previously cannibalized other larvae subsequently exhibited higher cannibalism rates than those larvae that had not cannibalized. In conclusion, low nitrogen, high larval density, and previous cannibalism experience are important factors contributing to gypsy moth larval cannibalism. Future studies are needed to estimate benefits to larvae, and to more closely approximate field conditions.     

Phenological asynchrony between host plant and gypsy moth reduces insect gut microbiota and susceptibility to Bacillus thuringiensis

The phenological synchrony between the emergence of overwintering herbivorous insects and the budding of host plants is considered a crucial factor in the population dynamics of herbivores. However, the mechanisms driving the interactions between the host plant, herbivores, and their pathogens are often obscure. In the current study, an artificially induced phenological asynchrony was used to investigate how the asynchrony between silver birch Betula pendula and gypsy moth Lymantria dispar affects the immunity of the insect to bacteria, its susceptibility to the entomopathogenic bacteria Bacillus thuringiensis, and the diversity in its midgut microbiota. The lysozyme‐like activity in both the midgut and hemolymph plasma and the nonspecific esterase activity and antimicrobial peptide gene expression in the midgut were studied in both noninfected and B. thuringiensis‐infected larvae. Our results provide the first evidence that phenologically asynchronous larvae are less susceptible to B. thuringiensis infection than phenologically synchronous larvae, and our results show that these effects are related to the high basic levels and B. thuringiensis‐induced levels of lysozyme‐like activities. Moreover, a 16S rRNA analysis revealed that dramatic decreases in the diversity of the larval gut bacterial consortia occurred under the effect of asynchrony. Larvae infected with B. thuringiensis presented decreased microbiota diversity if the larvae were reared synchronously with the host plant but not if they were reared asynchronously. Our study demonstrates the significant effect of phenological asynchrony on innate immunity‐mediated interactions between herbivores and entomopathogenic bacteria and highlights the role of nonpathogenic gut bacteria in these interactions.     

Effects of increased atmospheric CO2 on nutritional contents in poplar (Populus pseudo-simonii [Kitag.]) tissues and larval growth of gypsy moth (Lymantria dispar)

Changes in the concentrations of phytochemical compounds usually occur when plants are grown under elevated atmospheric CO₂. CO₂-induced changes in foliar chemistry tend to reduce leaf quality and may further affect insect herbivores. Increased atmospheric CO₂ also has a potential influence on decomposition because it causes variations in chemical components of plant tissues. To investigate the effects of increased atmospheric CO₂ on the nutritional contents of tree tissues and the activities of leaf-chewing forest insects, samples of Populus pseudo-simonii [Kitag.] grown in open-top chambers under ambient and elevated CO₂ (650 μmol mol^-1) conditions were collected for measuring concentrations of carbon, nitrogen, C : N ratio, soluble sugar and starch in leaves, barks, coarse roots (>2 mm in diameter) and fine roots (<2 mm in diameter). Gypsy moth (Lymantria dispar) larvae were reared on a single branch of experimental trees in a nylon bag with 1 mm 1 mm grid. The response of larval growth was observed in situ. Elevated CO₂ resulted in significant reduction in nitrogen concentration and increase in C : N ratio of all poplar tissues. In all tissues, total carbon contents were not affected by CO₂ treatments. Soluble sugar and nonstructural carbohydrate (TNC) in the poplar leaves significantly increased with CO₂ enrichment, whereas starch concentration increased only on partial sampling dates. Carbohydrate concentration in roots and barks was generally not affected by elevated CO₂, whereas soluble sugar contents in fine roots decreased in response to elevated CO₂. When second instar gypsy moth larvae consuming poplars grew under elevated CO₂ for the first 13 days, their body weight was 30.95% lower than that of larvae grown at ambient CO₂, but no significant difference was found when larvae were fed in the same treatment for the next 11 days. Elevated atmospheric CO₂ had adverse effects on the nutritional quality of Populus pseudo-simonii [Kitag.] tissues and the resultant variations in foliar chemical components had a significant but negative effect on the growth of early instar gypsy moth larvae.     

Facilitation between invasive herbivores: hemlock woolly adelgid increases gypsy moth preference for and performance on eastern hemlock

1. Interactions between invertebrate herbivores with different feeding modes are common on long-lived woody plants. In cases where one herbivore facilitates the success of another, the consequences for their shared host plant may be severe. Eastern hemlock (Tsuga canadensis), a canopy-dominant conifer native to the eastern U.S., is currently threatened with extirpation by the invasive stylet-feeding hemlock woolly adelgid (Adelges tsugae). The effect of adelgid on invasive hemlock-feeding folivores remains unknown. 2. This study evaluated the impact of feeding by hemlock woolly adelgid on gypsy moth (Lymantria dispar) larval preference for, and performance on, eastern hemlock. To assess preference, 245 field-grown hemlocks were surveyed for gypsy moth herbivory damage and laboratory paired-choice bioassays were conducted. To assess performance, gypsy moth larvae were reared to pupation on adelgid-infested or uninfested hemlock foliage, and pupal weight, proportional weight gain, and larval period were analysed. 3. Adelgid-infested hemlocks experienced more gypsy moth herbivory than did uninfested control trees, and laboratory tests confirmed that gypsy moth larvae preferentially feed on adelgid-infested hemlock foliage. Gypsy moth larvae reared to pupation on adelgid-infested foliage gained more weight than larvae reared on uninfested control foliage. 4. These results suggest that the synergistic effect of adelgid and gypsy moth poses an additional threat to eastern hemlock that may increase extirpation risk and ecological impact throughout most of its range.     

Effects of elevated CO2 on feeding preference and performance of the gypsy moth (Lymantria dispar) larvae

Multiple‐choice assays were conducted in the laboratory to examine the effect of CO₂‐induced changes in leaf quality on growth, nutritional indices and preferences of the gypsy moth (Lymantria dispar) larvae. The tested tree species, one‐year‐old aspen (Populus pseudo‐simonii Kitag.), two‐year‐old birch (Betula platyphylla) and three‐year‐old oak (Quercus mongolica Fisch.) were transplanted to open‐top chambers at ambient or elevated CO₂ (650 ppm) concentrations in May 2005. The present study was conducted in 2006. Leaves from the upper and lower crowns of each tree species were sampled and analysed, respectively. On both measurement dates (24 July 2006 and 16 August 2006), there were significant CO₂ effects on levels of condensed tannin, starch, nitrogen and C : N ratio. But only on the latter date, were the concentrations of total phenolics, soluble sugar, carbon and total non‐structural carbohydrates significantly affected by elevated CO₂. Leaf dry weight content and specific leaf weight were almost completely unaffected by CO₂ enrichment. Gypsy moth larvae exhibited a clear selectivity for tested leaf types (leaves from the upper and lower crowns of each tree species) even in their early instar stage, with the upper leaves of P. pseudo‐simonii being the most preferable and the lower leaves of Q. mongolica being the least preferred. The changes in leaf quality significantly reduced larval growth and altered partial indices of insect performance (e.g., relative growth rate and relative consumption rate). However, at least in this short‐term choice feeding assay (13 days), the CO₂‐induced changes in leaf quality had no significant effects on food preference behaviour of the gypsy moth larvae, neither within the limited range of host plants nor within the leaves at different canopy heights of the same tree species.     

Genetic determination of tannins and herbivore resistance in <Emphasis Type="Italic">Quercus ilex</Emphasis>

Genetic variability of trees influences the chemical composition of tissues. This determines herbivore impact and, consequently, herbivore performance. We evaluated the independent effects of plant genotype and provenance on the tannin content of holm oak (Quercus ilex) and their consequences for herbivory and performance of gypsy moth (Lymantria dispar) larvae. Oak seedlings of 48 open-pollinated families from six populations were grown in a common garden in central Spain. Half the plants were subjected to defoliation by gypsy moth larvae and the other half were destructively sampled for chemical analysis. Tannin content of leaves did not differ significantly among populations but differed significantly among families. Estimates of heritability (h ²) and quantitative genetic differentiation among populations for tannin content (Q _ST) were 0.83 and 0.12, respectively. Defoliation was not related to the tannin content of plants nor to spine and trichome densities of leaves, although positive family–mean associations were observed between defoliation and both seed weight and plant height (P < 0.003). Among the oak populations, differential increase in larval weight gain with defoliation was observed. Leaf tannin content in Q. ilex is genetically controlled but does not influence defoliation or predict performance of the larvae. Different efficiencies of food utilisation depending on the oak genotypes indicate that other plant traits are influencing the feeding patterns and fitness of L. dispar and consequent population dynamics.     

Effects of CO2-mediated changes in paper birch and white pine chemistry on gypsy moth performance

We examined the effects of CO₂-mediated changes in the foliar chemistry of paper birch (Betula papyrifera) and white pine (Pinus strobus) on performance of the gypsy moth (Lymantria dispar). Trees were grown under ambient or enriched CO₂ conditions, and foliage was subjected to plant chemical assays and insect bioassays. Enriched CO₂ atmospheres reduced foliar nitrogen levels and increased condensed tannin levels in birch but not in pine. Foliar carbohydrate concentrations were not markedly altered by CO₂ environment. Gypsy moth performance was significantly affected by CO₂ level, species, and the CO₂ x species interaction. Under elevated CO₂ conditions, growth was reduced for larvae fed birch, while development was prolonged for larvae fed pine. Although gypsy moths performed better overall on birch than pine, birch-fed larvae were influenced more by CO₂-mediated changes in host quality.     

Competition between the gypsy moth, Lymantria dispar, and the northern tiger swallowtail, Papilio canadensis: interactions mediated by host plant chemistry, pathogens, and parasitoids

The gypsy moth, Lymantria dispar, and the northern tiger swallowtail, Papilio canadensis, overlap geographically as well as in their host ranges. Adult female swallowtails are incapable of distinguishing between damaged and undamaged leaves, and the opportunities for competition between these two species are numerous. We designed field and laboratory experiments to look for evidence of indirect competition between P. canadensis and L. dispar larvae. Swallowtail caterpillars were reared in the laboratory on leaves from gypsy-moth-defoliated and undefoliated trees to explore host-plant effects. We tested for pathogen-mediated interactions by rearing swallowtail larvae on both sterilized and unsterilized leaves from defoliated and undefoliated sources. In addition, we measured the effects of known gypsy moth pathogens, as well as gypsy moth body fluids, on the growth and survival of swallowtail larvae. Field experiments were designed to detect the presence of parasitoid-mediated competition, as well: we recorded parasitism of swallowtail caterpillars placed in the field either where there were no gypsy moth larvae present, or where we had artificially created dense gypsy moth populations. We found evidence that swallowtails were negatively affected by gypsy moths in several ways: defoliation by gypsy moths depressed swallowtail growth rate and survival, whether leaves were sterilized or not; sterilization significantly reduced the effect of defoliation, and gypsy moth body fluids proved lethal; and swallowtail caterpillars suffered significantly increased rates of parasitism when they were placed in the field near gypsy moth infestations.     

Accumulation of Cd and Pb in various body parts,organs and tissues of Lymantria dispar asiatica (Lepidoptera: Erebidae)

To investigate the accumulation patterns of Cd and/or Pb in various body parts, organs and tissues of the Asian gypsy moth (Lymantria dispar) larvae under Cd and/or Pb stress, Cd and Pb treated artificial diets were used to feed the larvae in the current study. These larval body parts/organs/tissues included the heads, integuments (body walls), alimentary canals, fat bodies and hemolymphs. Our results showed that under Cd and/or Pb stress, their accumulations in larval body parts/organs/tissues were significantly higher than those in the control, with the amounts of tested metals in the fat bodies and hemolymphs and alimentary canals being significantly higher than those in the heads and integuments. Under the single Cd (0, 7.5, 10.5 mg/kg) or Pb stress (0, 55, 90, 125 mg/kg), the accumulations of these heavy metals were positively correlated with their concentrations in diets. Under the combined stress (Cd × Pb), the Cd accumulation at the lower Pb concentrations was higher than that at higher Pb concentrations for the body parts/organs/tissues, similar effects of Cd concentrations on Pb accumulations were also recorded. These results clearly showed that Cd and/or Pb were accumulated in various body parts/organs/tissues at different levels under the heavy metal stress. And accumulations of Pb/Cd were positively correlated with their concentrations in the diets under the single stress. Under the combined stresses, Cd and Pb had synergistic effects at low concentrations whereas antagonistic effects at high concentrations. The accumulations of Cd and/or Pb in the gypsy moth larvae affects normal physiological and biochemical functions, and thus affects their growth and development.     

Variation in the ability of larvae of phytophagous insects to develop on evolutionarily unfamiliar plants: a study with gypsy moth Lymantria dispar and Eucalyptus

• 1 By examining variation in the abilities of polyphagous insects to develop on host plants with secondary metabolites that they have never encountered previously, we may be able to gain some insights into the nature of evolution of biochemical mechanisms to process plant secondary metabolites by phytophagous insects.
• 2 The present study aimed to examine variation in the ability of gypsy moth larvae Lymantria dispar (Lymantriidae) to complete development on different species of the plant genus Eucalyptus (Myrtaceae). Leaves of at least some Eucalyptus species contain formylated phloroglucinol derivatives. These are secondary metabolites that are evolutionarily unfamiliar to the gypsy moth.
• 3 Larvae of gypsy moth showed extremely variable responses in larval performance between Eucalyptus species, between individual trees within host plant species, between moth populations, and between individuals within moth populations.
• 4 Larval survivorship was in the range 0–94%, depending on the host. Failure of at least some larvae to complete development on some Eucalyptus species indicates that gypsy moth larvae have a limited ability to process secondary metabolites in eucalypt leaves.
• 5 At least some individuals, however, appear to already possess biochemical mechanisms that process the secondary metabolites in leaves of Eucalyptus species, and therefore the abilities of larvae to complete development on phylogenetically and chemically unfamiliar hosts are already present before the gypsy moth encounters these potential hosts.

     

Effects of long-term simulated acid rain on a plant–herbivore interaction

Anthropogenic pollution causes oxidative stress in plants and reactive oxygen species (ROS) are diminished by antioxidative enzymes and small molecular antioxidants. Pollution may also affect the performance of plant-eating animals by increasing or decreasing their performance. The effects of pollution cannot be fully understood without knowledge of how pollution affects the interactions with the third trophic level, namely natural enemies and diseases of herbivores. In this study, we examined how long-term (19 yr) acid rain pollution affects (i) the oxidative responses in mountain birch foliage and (ii) the growth and immune responses of autumnal moth larvae. We found that pollution caused a 50% increase (p<0.05) in the peroxidase activities (PODs) in birch leaves whereas polyphenoloxidase (PPO) or catalase (CAT) activities were not affected, suggesting that PODs play an important role in the quenching of the oxidative stress in birches. In polluted trees, phenoloxidases probably acted as antioxidative not prooxidative enzymes, which was shown as positive relations between enzyme activities (PPO, CAT) and larval performance (pupal weights). Although acid rain pollution did not have any direct effect on either pupal weight or the length of larval period, the stronger acid rain treatment reduced slightly (6% in females) the encapsulation response of pupae. A decrease of this magnitude might be too small to have measurable effects on the incidence of moth outbreaks.     

Folivorous larvae on flowers: do autumnal moths benefit from catkins of the mountain birch?

Immature leaves of birches and other deciduous trees and shrubs are the principal food resource for the larvae of the autumnal moth, Epirrita autumnata (Borkhausen) (Lepidoptera: Geometridae). However, the larvae of this geometrid, which displays pronounced population cycles and causes massive forest defoliations in northernmost Europe, are also frequently found on the catkins of mountain birch, Betula pubescens ssp. czerepanovii (Orlova) Hämet-Ahti (Betulaceae). To examine whether autumnal moth larvae visit birch catkins for their high nutritional value, we conducted two growth experiments in which the larval diet was supplemented with male catkins, and a single test where female catkins were used. We found that the presence of male catkins in the larval diet had a positive effect on the larvae, in relation to their pupal mass (reflecting life span fecundity), survival probability, as well as the duration of the larval period. The presence of female catkins in the diet did not affect larval performance. Our findings suggest that young autumnal moth larvae actively move to male catkins, which provide a nutritional benefit and thus lead to a better performance compared with those feeding elsewhere. Another possible benefit for young larvae choosing the catkins instead of leaves may involve predator or parasitoid avoidance.     

Cyclic fluctuations in arylphorin,the principal serum storage protein of Lymantria dispar,indicate multiple roles in development

Arylphorin (Ap) was isolated and partially characterized from larval serum of the gypsy moth, Lymantria dispar (Lepidoptera: Lymantriidae). Non-denaturing and SDS-polyacrylamide gel electrophoresis suggest a 440,000 Da hexamer composed of nonidentical subunits of M_r 73,000 and 80,000. Tyrosine and phenylalanine comprise 15.6% of the total protein amino acid. L. dispar Ap is immunologically related to Manduca sexta Ap. Hemolymph Ap concentrations throughout the larval stadia were determined using quantitative immunoelectrophoresis. Cyclic fluctuations in hemolymph concentrations are correlated with each molting cycle. A general increase in Ap concentration during each intermolt is followed by a sharp decline between apolysis and ecdysis. The last days of each instar were found to be the best time to sample protein titer with minimum variance. Hemolymph space, estimated by measuring the dilution of an injected foreign protein, is a constant 28% (v/w) of body weight. Total serum Ap per animal was calculated. A divergent allometric relationship between Ap accumulation and weight gain throughout the larval period results in the ultimate domination of the serum protein profile by Ap in the ultimate larval instar of the female in particular. An additional instar of Ap accumulation in the female gypsy moth is suggested to compensate for the lack of a predominantly female-specific storage protein in this species.     

Intraspecific variation in aspen phytochemistry: effects on performance of gypsy moths and forest tent caterpillars

Individual quaking aspen trees vary greatly in foliar chemistry and susceptibility to defoliation by gypsy moths and forest tent caterpillars. To relate performance of these insects to differences in foliar chemistry, we reared larvac from egg hatch to pupation on leaves from different aspen trees and analyzed leaf samples for water, nitrogen, total nonstructural carbohydrates, phenolic glycosides, and condensed tannins. Larval performance varied markedly among trees. Pupal weights of both species were strongly and inversely related to phenolic glycoside concentrations. In addition, gypsy moth performance was positively related to condensed tannin concentrations, whereas forest tent caterpillar pupal weights were positively associated with leaf nitrogen concentrations. A subsequent study with larvae fed aspen leaves supplemented with the phenolic glycoside tremulacin confirmed that the compound reduces larval performance. Larvae exhibited increased stadium durations and decreased relative growth rates and food conversion efficiencies as dietary levels of tremulacin increased. Differences in performance were more pronounced for gypsy moths than for forest tent caterpillars. These results suggest that intraspecific variation in defensive chemistry may strongly mediate interactions between aspen, gypsy moths and forest tent caterpillars in the Great Lakes region, and may account for differential defoliation of aspen by these two insect species.     

Leaf age effects of elevated CO2-grown white oak leaves on spring-feeding lepidopterans

Folivorous insect responses to elevated CO₂-grown tree species may be complicated by phytochemical changes as leaves age. For example, young expanding leaves in tree species may be less affected by enriched CO₂-alterations in leaf phytochemistry than older mature leaves due to shorter exposure times to elevated CO₂ atmospheres. This, in turn, could result in different effects on early vs. late instar larvae of herbivorous insects. To address this, seedlings of white oak (Quercus alba L.), grown in open-top chambers under ambient and elevated CO₂, were fed to two important early spring feeding herbivores; gypsy moth (Lymantria dispar L.), and forest tent caterpillar (Malacosoma disstria Hübner). Young, expanding leaves were presented to early instar larvae, and older fully expanded or mature leaves to late instar larvae. Young leaves had significantly lower leaf nitrogen content and significantly higher total nonstructural carbohydrate:nitrogen ratio as plant CO₂ concentration rose, while nonstructural carbohydrates and total carbon-based phenolics were unaffected by plant CO₂ treatment. These phytochemical changes contributed to a significant reduction in the growth rate of early instar gypsy moth larvae, while growth rates of forest tent caterpillar were unaffected. The differences in insect responses were attributed to an increase in the nitrogen utilization efficiency (NUE) of early instar forest tent caterpillar larvae feeding on elevated CO₂-grown leaves, while early instar gypsy moth larval NUE remained unchanged among the treatments. Later instar larvae of both insect species experienced larger reductions in foliage quality on elevated CO₂-grown leaves than earlier instars, as the carbohydrate:nitrogen ratio of leaves substantially increased. Despite this, neither insect species exhibited changes in growth or consumption rates between CO₂ treatments in the later instar. An increase in NUE was apparently responsible for offsetting reduced foliar nitrogen for the late instar larvae of both species.     

Microbial synergists pathogenic to Lymantria dispar: Chitinolytic and fermentative bacterial interactions

A chitinolytic bacterium isolated from a healthy gypsy moth, Lymantria dispar, larva was shown to be pathogenic to larvae when administered per os after growth on chitin broth. However, the chitin hydrolase produced by this isolate had a pH optimum for activity of 5.5 and the high alkalinity in the mesenteron of L. dispar larvae severely limited chitinolysis of the midgut lining. Fermentative, nonpathogenic, acid-producing bacteria isolated from healthy gypsy moth larvae effectively lowered larval mesenteron pH when administered per os and the combination of fermentative isolates with a crude culture aliquot of the chitinolytic strain produced a synergistic increase in mortality over either dose administered by itself. Increased mortality was also observed for most fermentative strains when they were combined with crude supernatants of centrifuged cultures of the chitinolytic strain, although these combinations proved less effective than when fermenters were added to the whole-culture aliquots of the chitinolytic strain. In vitro studies showed that other bacteria isolated from environments foreign to that of the gypsy moth could ferment carbohydrates with acid production at an alkaline pH; however, in vivo studies demonstrated that these bacteria were incapable both of poising larval midgut pH and of enhancing mortality when added to chitinolytic bacteria.