Genetic control and evolutionary potential of a constitutive resistance mechanism against the spruce budworm (Choristoneura fumiferana) in white spruce (Picea glauca).

Insect herbivory may drive evolution by selecting for trees with heritable resistance against defoliation. The spruce budworm (Choristoneura fumiferana, SBW) is a highly damaging forest insect pest that can affect population structure of white spruce (Picea glauca) in North America. Resistance against SBW was recently described in white spruce and was linked to three constitutive resistance biomarkers: the phenolic compounds piceol and pungenol, and expression of a beta-glucosidase encoding gene (Pgßglu-1). We investigated the phenotypic variability and heritability of these resistance biomarkers and of picein, the precursor of piceol, in the foliage of 874 trees belonging to 33 full-sib families and 71 clonal lines under evaluation in seven field locations in Eastern Canada. We aimed to (i) determine their genetic control, (ii) estimate the genetic and phenotypic correlations among defense biomarkers, and (iii) determine whether their constitutive levels are associated with detrimental trade-offs on growth. Quantitative genetics analyses indicated that all four traits are moderately to highly heritable. The full-sib and clonal analyses showed that additive and non-additive genetic effects play major and minor roles, respectively. Positive genetic and phenotypic correlations between resistance biomarkers and primary growth indicated that there is no trade-off between total height and height increment and resistance traits, contradicting the GDBH (Growth Differentiation Balance Hypothesis). Our findings about the predominant additive genetic basis of the resistance biomarkers show that adaptive evolution of white spruce natural populations to resist to SBW is possible and that potentially important gains could also be expected from artificial selection.

How does synchrony with host plant affect the performance of an outbreaking insect defoliator?

Phenological mismatch has been proposed as a key mechanism by which climate change can increase the severity of insect outbreaks. Spruce budworm (Choristoneura fumiferana) is a serious defoliator of North American conifers that feeds on buds in the early spring. Black spruce (Picea mariana) has traditionally been considered a poor-quality host plant since its buds open later than those of the preferred host, balsam fir (Abies balsamea). We hypothesize that advancing black spruce budbreak phenology under a warmer climate would improve its phenological synchrony with budworm and hence increase both its suitability as a host plant and resulting defoliation damage. We evaluated the relationship between tree phenology and both budworm performance and tree defoliation by placing seven cohorts of budworm larvae on black spruce and balsam fir branches at different lags with tree budburst. Our results show that on both host plants, spruce budworm survival and pupal mass decrease sharply when budbreak occurs prior to larval emergence. By contrast, emergence before budbreak decreases survival, but does not negatively impact growth or reproductive output. We also document phytochemical changes that occur as needles mature and define a window of opportunity for the budworm. Finally, larvae that emerged in synchrony with budbreak had the greatest defoliating effect on black spruce. Our results suggest that in the event of advanced black spruce phenology due to climate warming, this host species will support better budworm survival and suffer increased defoliation.

Expression of the ß-glucosidase gene Pgßglu-1 underpins natural resistance of white spruce against spruce budworm.

Periodic outbreaks of spruce budworm (SBW) affect large areas of ecologically and economically important conifer forests in North America, causing tree mortality and reduced forest productivity. Host resistance against SBW has been linked to growth phenology and the chemical composition of foliage, but the underlying molecular mechanisms and population variation are largely unknown. Using a genomics approach, we discovered a ß-glucosidase gene, Pgßglu-1, whose expression levels and function underpin natural resistance to SBW in mature white spruce (Picea glauca) trees. In phenotypically resistant trees, Pgßglu-1 transcripts were up to 1000 times more abundant than in non-resistant trees and were highly enriched in foliage. The encoded PgßGLU-1 enzyme catalysed the cleavage of acetophenone sugar conjugates to release the aglycons piceol and pungenol. These aglycons were previously shown to be active against SBW. Levels of Pgßglu-1 transcripts and biologically active acetophenone aglycons were substantially different between resistant and non-resistant trees over time, were positively correlated with each other and were highly variable in a natural white spruce population. These results suggest that expression of Pgßglu-1 and accumulation of acetophenone aglycons is a constitutive defence mechanism in white spruce. The progeny of resistant trees had higher Pgßglu-1 gene expression than non-resistant progeny, indicating that the trait is heritable. With reported increases in the intensity of SBW outbreaks, influenced by climate, variation of Pgßglu-1 transcript expression, PgßGLU-1 enzyme activity and acetophenone accumulation may serve as resistance markers to better predict impacts of SBW in both managed and wild spruce populations.

Effect of temperature and tree species on damage progression caused by whitespotted sawyer (Coleoptera: Cerambycidae) larvae in recently burned logs.

The whitespotted sawyer, Monochamus scutellatus scutellatus (Say) (Coleoptera: Ce-rambycidae), is one of the most damaging wood-boring insects in recently burned boreal forests of North America. In Canada, salvage logging after wildfire contributes to maintaining the timber volume required by the forest industry, but larvae of this insect cause significant damage that reduces the economic value of lumber products. This study aimed to estimate damage progression as a function of temperature in recently burned black spruce (Picea mariana (Miller) Britton, Sterns, and Poggenburg) and jack pine (Pinus banksiana Lambert) trees. Using axial tomographic technology, we modeled subcortical development and gallery depth progression rates as functions of temperature for both tree species. Generally, these rates were slightly faster in black spruce than in jack pine logs. Eggs laid on logs kept at 12 degrees C did not hatch or larvae were unable to establish themselves under the bark because no larval development was observed. At 16 degrees C, larvae stayed under the bark for > 200 d before penetrating into the sapwood. At 20 degrees C, half of the larvae entered the sapwood after 30-50 d, but gallery depth progression stopped for approximately 70 d, suggesting that larvae went into diapause. The other half of the larvae entered the sapwood only after 100-200 d. At 24 and 28 degrees C, larvae entered the sapwood after 26-27 and 21 d, respectively. At 28 degrees C, gallery depth progressed at a rate of 1.44 mm/d. Temperature threshold for subcortical development was slightly lower in black spruce (12.9 degrees C) than in jack pine (14.6 degrees C) and it was 1 degrees C warmer for gallery depth progression for both tree species. These results indicate that significant damage may occur within a few months after fire during warm summers, particularly in black spruce, which highlights the importance of beginning postfire salvage logging as soon as possible to reduce economic losses.

Intra-tree variation in foliage quality drives the adaptive sex-biased foraging behaviors of a specialist herbivore.

Some herbivorous insects enhance their fitness using foraging strategies that allow them to find and colonize the best of available resources within heterogeneous plants. The yellowheaded spruce sawfly, Pikonema alaskensis (Roh.) (Hymenoptera: Tenthredinidae), is a common defoliator that oviposits and feeds on the developing foliage of young open-grown black spruce [Picea mariana (Mills) B.S.P.]. While female and male eggs are both laid throughout the crown, most eggs laid in the upper crown are female, and more female than male late-instar larvae disperse acropetally, from the lower and mid-crown to the upper crown, to complete juvenile development. Here we present results from 4 years of manipulative sleeve-cage experiments that were carried out to evaluate the hypothesis that sex-biased oviposition-site selection and acropetal dispersal by P. alaskensis are adaptive responses to intra-tree variation in foliage quality. Survival and proportion of survivors that were female were either the same (2 years) or significantly higher (2 years) for groups placed in the lower or mid-crown during early instars, and then transferred acropetally during late instars to complete development in the upper crown, compared with those forced to feed exclusively in the lower crown. This suggests that females benefited most from acropetal dispersal. Sex ratios of survivors that had been forced to develop exclusively in the upper crown were usually more female biased than those of survivors that developed exclusively in the lower crown, suggesting higher survival for female than for male larvae emerging from eggs laid in the upper crown. Sex-biased egg allocation and larval dispersal appear to increase the survival of P. alaskensis by accounting for differential effects on male versus female larvae of phenology-independent temporal and spatial variations in the quality of foliage within the heterogeneous crown of black spruce.

Host Tree Species Affects Spruce Budworm Winter Survival.

With current trends in global warming, it has been suggested that spruce budworm outbreaks may spread to northern parts of the boreal forest. However, the major constraints for a northward expansion are the availability of suitable host trees and the insect winter survival capacity. This study aimed to determine the effect of larval feeding on balsam fir, white spruce and black spruce on various spruce budworm life history traits of both the parental and the progeny generations. Results indicated that the weight of the overwintering larval progeny and their winter survival were influenced by host tree species on which larvae of the parental generation fed. White spruce was the most suitable host for the spruce budworm, producing the heaviest pupae and the heaviest overwintering larvae while black spruce was the least suitable, producing the smallest pupae and the smallest overwintering progeny. Overwintering larvae produced by parents that fed on black spruce also suffered higher winter mortality than individuals coming from parents that fed on balsam fir or white spruce. With current trends in global warming, spruce budworm is expected to expand its range to northern boreal forests where black spruce is the dominant tree species. Such northern range expansion might not result in outbreaks if low offspring winter survival on black spruce persist.

Hydroxyacetophenone defenses in white spruce against spruce budworm.

We review a recently discovered white spruce (Picea glauca) chemical defense against spruce budworm (Choristoneura fumiferana) involving hydroxyacetophenones. These defense metabolites detected in the foliage accumulate variably as the aglycons, piceol and pungenol, or the corresponding glucosides, picein and pungenin. We summarize current knowledge of the genetic, genomic, molecular, and biochemical underpinnings of this defense and its effects on C. fumiferana. We present an update with new results on the ontogenic variation and the phenological window of this defense, including analysis of transcript responses in P. glauca to C. fumiferana herbivory. We also discuss this chemical defense from an evolutionary and a breeding context.

Detoxification of host plant phenolic aglycones by the spruce budworm.

This study examines the post-ingestive fate of two host-plant derived small-molecule phenolics (the acetophenones piceol and pungenol) that have previously been shown to be toxic to the outbreaking forest pest, spruce budworm (Choristoneura fumiferana). We test first whether these compounds are transformed during passage through the midgut, and second whether the budworm upregulates activity of the detoxification enzyme glutathione-s-transferase (GST) in response to feeding on these compounds. Insects were reared on either foliage or artificial diet to the fourth instar, when they were transferred individually to one of two treatment diets, either control or phenolic-laced, for approximately 10 days, after which midguts were dissected out and used for Bradford soluble protein and GST enzyme activity analysis. Frass was collected and subjected to HPLC-DAD-MS. HPLC showed that the acetophenones do not autoxidize under midgut pH conditions, but that glucose- and glutathione- conjugates are present in the frass of insects fed the phenolic-laced diet. GST enzyme activity increases in insects fed the phenolic-laced diet, in both neutral pH and alkaline assays. These data show that the spruce budwom exhibits counter-adaptations to plant phenolics similar to those seen in angiosperm feeders, upregulating an important detoxifying enzyme (GST) and partially conjugating these acetophenones prior to elimination, but that these counter-measures are not totally effective at mitigating toxic effects of the ingested compounds in the context of our artifical-diet based laboratory experiment.

Phenotypic variation in food utilization in an outbreak insect herbivore.

The effects of nutrition may have subtantial impact on insect evolution by shaping different components of phenotypes. The key to undestanding this evolutionary process is to know how nutritional condition affects additive and nonadditive components of the phenotype. However, this is poorly understood in outbreaking insects. We investigated the additive and nonadditive variation present in food utilization traits in spruce budworm individuals subjected to chronic nutritional stress. A total of 160 full-sib families of spruce budworm (Choristoneura fumiferana Clem.) were raised under laboratory conditions, feeding on 2 diets (high and low energy) during 3 generations. Variables tested were pupal mass, consumption rate (RCR), growth rate (RGR), approximate digestibility (AD), the efficiency of conversion of digested food (ECD) and the efficiency of conversion of ingested food (ECI). Our results show that all traits tested presented a high percentage of nonadditive effects that modulate phenotype expression. We found a significant impact of family × diet interaction on pupal mass, RGR and ECD. Furthermore, these traits exhibited the greatest heritability. There was no evidence of presence of maternal effects. The results revealed that food utilization traits may evolve through epigenetics effects, such as phenotypic plasticity. This information can be used by modellers to improve forecast of spruce budworm population dynamics.

Developmental polymorphism in a Newfoundland population of the hemlock looper, Lambdina fiscellaria (Lepidoptera: Geometridae).

The hemlock looper [Lambdina fiscellaria (Guenée)], a widespread and highly polyphagous Geometridae, is considered one of the most economically important defoliators of North American coniferous forests. Variations in the number of larval instars between geographic populations of this species have been previously reported in the literature. However, whether such developmental polymorphism occurs within a given population is unknown. In this study, we report the presence of both four and five larval instar individuals within a population of hemlock looper in Newfoundland when reared on balsam fir. For both sexes, the majority of individuals reared on balsam fir shoots went through four larval instars, but more than one third of the females (35.3%) went through five larval instars. Females with four larval instars developed faster and had smaller pupal weight than females with five larval instars. However, a growth-related index (weight gain per unit of time) was similar for the two ecotypes (four or five larval instars). No significant difference was observed between the two ecotypes in terms of reproductive capacity (fecundity and egg size). We also found significant differences in life history traits between males and females. Results indicate that developmental polymorphism, in this case, the variation in the number of larval instars, might provide some adaptive attributes that allowed exploitation of a broader ecological niche.

Diapause disruption with tebufenozide for early-instar control of the spruce budworm, Choristoneura fumiferana.

In North America, the eastern spruce budworm, Choristoneura fumiferana Clem., is an important coniferous pest against which tebufenozide has proven effective as a control product. By acting as an ecdysone agonist, tebufenozide can induce precocious moulting in late (fifth-sixth) instars but can also be carried over to the next generation owing to its persistence on foliage. The authors conducted laboratory experiments on first-instar larvae treated with tebufenozide dissolved in acetone. Larvae exposed to doses equal to or above 0.1 microg cm(-2) displayed precocious moulting in the second instar after hibernaculum spinning, which effectively disrupted diapause. Larger doses induced moulting in first instars. Evidence is provided that this dose-response difference is related to whether or not an effective dose of tebufenozide is ingested by the first instar prior to the peak of moulting hormone (20-hydroxyecdysone) in first instars. Doses ineffective to kill first instars are carried over to the second instar, where they induce a precocious moult. This type of response to tebufenozide is dependent on the presence of a moulting machinery (the EcR-USP receptor complex) that is ready for ecdysone transduction. Interestingly, ecdysone levels are low in second instars, as measured by a radioimmunoassay, which suggests that diapause in spruce budworm is maintained by a suppression of ecdysone production. Thus, diapause disruption by tebufenozide may well provide an alternative control strategy for this important pest.

Spruce budworm feeding and oviposition are stimulated by monoterpenes in white spruce epicuticular waxes.

Monoterpenes, source of the distinctive odor of conifers, are generally considered plant defensive compounds. However, they are also known to act as long-range insect attractants, as they are volatile and permeate forest airspaces. Moreover, they are lipid soluble and can be absorbed into plant epicuticular waxes. We test their role in short-range host plant choice by both adult females and larvae of a folivorous forest pest (Choristoneura fumiferana). We conducted laboratory assays testing the responses of Eastern spruce budworm to an artificial monoterpene mix (α-pinene, ß-pinene, limonene, myrcene) and to white spruce (Picea glauca) epicuticular waxes in closed arenas. Ovipositing females preferred filter paper discs treated with P. glauca waxes to controls, and preferred the waxes + monoterpenes treatment to waxes alone. However, females showed no preference between the monoterpene-treated disc and the control when presented without waxes. Feeding larvae prefered wax discs to control discs. They also consumed discs treated with realistic monoterpene concentrations and wax preferentially over wax-only discs, but showed no preference between extremely high monoterpene concentrations and wax-only controls. In an insect-free assay, P. glauca epicuticular wax decreased monoterpene volatilization. These results suggest that P. glauca waxes and realistic concentrations of monoterpenes are stimulatory to both egg-laying females and feeding larvae, and that their effects are synergistic.

Insect herbivory (Choristoneura fumiferana, Tortricidea) underlies tree population structure (Picea glauca, Pinaceae).

Variation in insect herbivory can lead to population structure in plant hosts as indicated by defence traits. In annual herbaceous, defence traits may vary between geographic areas but evidence of such patterns is lacking for long-lived species. This may result from the variety of selection pressures from herbivores, long distance gene flow, genome properties, and lack of research. We investigated the antagonistic interaction between white spruce (Picea glauca) and spruce budworm (SBW, Choristoneura fumiferana) the most devastating forest insect of eastern North America in common garden experiments. White spruces that are able to resist SBW attack were reported to accumulate the acetophenones piceol and pungenol constitutively in their foliage. We show that levels of these acetophenones and transcripts of the gene responsible for their release is highly heritable and that their accumulation is synchronized with the most devastating stage of SBW. Piceol and pungenol concentrations negatively correlate with rate of development in female SBW and follow a non-random geographic variation pattern that is partially explained by historical damage from SBW and temperature. Our results show that accumulation of acetophenones is an efficient resistance mechanism against SBW in white spruce and that insects can affect population structure of a long-lived plant.

Age-dependent fecundity and fertility life tables of the predator Brontocoris tabidus (Heteroptera: Pentatomidae) under field conditions.

Reproductive potential, longevity, life expectancy, and fertility life tables of Brontocoris tabidus (Signoret) (Heteroptera: Pentatomidae), a predator of lepidopteran defoliators in eucalyptus (Eucalyptus spp.) plantations, were studied in the field. After a 50-d preoviposition period (emergence of adults to the deposition of the first egg mass), ovipositional activity of B. tabidus continued until females died at 160 d. Females laid an average of 4.2 eggs per day and 601.1 eggs in a lifetime. Gross and net reproductive rates were 216.7 and 75.8 females, respectively. Generation time was 146.1 d, the period for doubling the population was 23.4 d, intrinsic rate was 0.03, and finite population increase was 1.03. Number of females per generation increased at 33.4 times. Results from our field studies indicate that B. tabidus has greater potential reproduction, oviposition period, and longevity than was expected from previous laboratory experiments. This suggests that B. tabidus has potential as a biological control agent to limit economically damaging pests in eucalyptus plantations.

Do Offspring of Insects Feeding on Defoliation-Resistant Trees Have Better Biological Performance When Exposed to Nutritionally-Imbalanced Food?

White spruce (Picea glauca (Moench) Voss) trees that are resistant or susceptible to spruce budworm (Choristoneura fumiferana (Clem.)) attack were identified in a southern Quebec plantation. Due to high mortality-induced selective pressures imposed by resistant trees on spruce budworm larvae, insects that survive on resistant trees exhibited greater biological performance than those on susceptible trees. We tested the hypothesis that this better biological performance is maintained across generations when progeny were subjected to nutritional stress. We collected pupae from resistant and susceptible trees (phenotype). Adults were reared under controlled laboratory conditions. Progeny were subsequently reared on two types of artificial diet (high vs. low quality). Low quality diet simulated food quality deterioration during outbreak conditions. Results confirmed that surviving insects collected from resistant trees have better performance than those from susceptible trees. Offspring performance (pupal mass, developmental time) was affected only by diet quality. These results suggest that adaptive advantages that would be acquired from parents fed on resistant trees are lost when progeny are exposed to nutritionally-imbalanced food, but the effects persist when larvae are fed a balanced diet. Offspring mortality, fecundity and fertility were positively influenced by parental origin (tree phenotype).

Effect of simulated fall heat waves on cold hardiness and winter survival of hemlock looper, Lambdina fiscellaria (Lepidoptera: Geometridae).

The hemlock looper (Lambdina fiscellaria) is an important pest of eastern Canadian forests. The ongoing climate warming could modify the seasonal ecology of this univoltine species that lays eggs at the end of summer and overwinters at this stage. Indeed, the increase in frequency and intensity of extreme climatic events such as fall heat waves could interfere with the winter metabolism of the hemlock looper. Moreover, the host plant quality, which influences the quantity of insect energetic reserves, the geographic origin of populations and the conditions prevailing during the cold acclimation period, could cause various responses of this pest to climate warming. The main objective of this study is to determine the impact of these factors on hemlock looper winter biology. In October 2010, hemlock looper eggs initially collected from two geographic areas in the province of Québec, and from parents reared on two host plants, were exposed to fall heat waves of different intensities during 5 consecutive days. Supercooling points and cryoprotectant levels were measured on eggs on four different dates in 2010-2011 and survival rate was measured in April 2011. Our results show that hemlock looper eggs have a very low supercooling point and high levels of trehalose, glucose and mannitol in September and November. However, there is no clear relationship between the concentration of these compounds and the decrease in supercooling points. Contents in trehalose, glucose and mannitol were significantly influenced by fall heat waves and by the origin of the population. Winter survival of eggs from the temperate population was negatively affected by strong heat waves while the boreal population was not affected. This study suggests that the metabolism and winter survival of temperate hemlock looper populations in Québec will be more affected by fall heat waves that will increase in frequency due to climate change, than boreal populations.

Larval feeding behaviour affects the impact of staminate flower production on the suitability of balsam fir trees for spruce budworm.

Laboratory rearing of spruce budworm, Choristoneura fumiferana, in conjunction with field rearing indicated that the feeding behaviour of the larvae, which is affected by the insect population density, significantly influenced the impact of balsam fir, Abies balsamea, staminate flowering on spruce budworm biology. At low budworm density, the production of pollen in the midcrown of host trees reduced the insect development time by 5 days without affecting pupal weight, fecundity and survival. However, at high budworm density, the small amount of current-year foliage produced by flowering branches forced old larvae (sixth instar) either to feed on 1-year-old foliage (backfeeding) or to move from the midcrown to the lower crown section where staminate flowers are absent and more current-year foliage is available. When old larvae fed on old foliage, they exhibited reduced pupal weight and fecundity without losing the advange in development time that they obtained from feeding on pollen during their early stages of development. On the other hand, when old larvae moved to the lower crown section, they avoided the negative effects of backfeeding but lost the advantage in development time that was gained from feeding on pollen. Results from this study indicated that the production of staminate flowers by balsam fir trees could have opposite effects on spruce budworm population dynamics depending upon the insect population density when flowering occurs.

Lethal and sublethal effects of single and double applications of Bacillus thuringiensis variety kurstaki on spruce budworm (Lepidoptera: Tortricidae) larvae.

We conducted laboratory experiments to examine the effects of single versus double exposures of spruce budworm, Choristoneura fumiferana (Lepidoptera: Tortricidae) female larvae to various concentrations of a Bacillus thuringiensis variety kurstaki (Btk) commercial formulation (Foray 48B). Our main objective was to document the vulnerability to Btk and the sublethal responses of fifth-instar larvae that survived from a first ingestion of Btk during their fourth stadium and to compare them with insects treated either during their fifth or fourth stadium only. As reported in the literature, fifth-instar larvae were more vulnerable than fourth-instar larvae, but only at low and medium concentrations. Fifth-instar larvae that had survived Btk ingestion during their fourth stadium were more vulnerable to a high concentration of Btk and had a shorter feeding inhibition period than those that had not been exposed during their fourth stadium. Compared with a single treatment at the fourth stadium, a double exposure to Btk further reduced the population by 20-30%, depending on the concentration applied. The second treatment also induced another feeding inhibition period and increased larval development time by 14%. The impact of the different treatments on pupal weight depended on whether treated insects exhibited supernumerary instars. In the absence of developmental polymorphism, a higher concentration, a late, or a double exposure to Btk significantly reduced pupal weight.

Bacillus thuringiensis subsp. kurstaki aerial spray prescriptions for balsam fir stand protection against spruce budworm (Lepidoptera: Tortricidae).

Although commercial formulations of Bacillus thuringiensis subsp. kurstaki (Btk) are being widely used in forest protection against lepidopteran defoliators, optimal application prescriptions have often yet to be worked out in detail. We conducted field experiments over a 6-yr period (1996-2001) in southwestern Québec to determine application prescriptions for optimal protection of balsam fir, Abies balsamea (L.), healthy stands against the spruce budworm, Choristoneura fumiferana (Clemens) (Lepidoptera: Tortricidae). At moderate larval densities (<30 larvae per 45-cm branch tip), similar foliage protection was achieved with one or two Btk applications of 30 billion international units per hectare (BIU/ha). When larval densities exceeded 30 larvae per branch tip, two successive applications of 30 BIU/ha significantly increased foliage protection. Whether the second application took place 5 or 10 d after the first spray did not affect treatment efficacy. Increasing the application dosage from 30 to 50 BIU/ha did not lead to better foliage protection against high larval densities, but the current standard dosage of 30 BIU/ha saved more foliage than 15 BIU/ha against moderate populations. The recommended dosage of 30 BIU can be applied in lower application volumes (1.5 liters/ha) by using a high-potency product (20 BIU/liter), because we did not observe a reduction in efficacy compared with the application of a lower potency product (12.7 BIU/liter) in 2.37 liters/ha. We also demonstrated that Btk can be applied much earlier in the season without compromising spray efficacy: there was no difference in treatment efficacy of double applications at 30 BIU/ha when the first spray was timed for early third, peak third, or early fourth instars.

Life-history consequences of chronic nutritional stress in an outbreaking insect defoliator.

Food shortage is a common situation in nature but little is known about the strategies animals use to overcome it. This lack of knowledge is especially true for outbreaking insects, which commonly experience nutritional stress for several successive generations when they reach high population densities. The aim of this study is to evaluate the life history consequences of chronic nutritional stress in the outbreaking moth Choristoneura fumiferana. Larvae were reared on two different artificial diets that emulate nutritional conditions larvae face during their natural population density cycle (low and medium quality artificial diets). After four generations, a subset of larvae was fed on the same diet as their parents, and another on the opposite diet. We explored larval life-history strategies to cope with nutritional stress, its associated costs and the influence of nutritional conditions experienced in the parental generation. We found no evidence of nutritional stress in the parental generation increasing offspring ability to feed on low quality diet, but the contrary: compared to offspring from parents that were fed a medium quality diet, larvae from parents fed a low quality diet had increased mortality, reduced growth rate and reduced female reproductive output. Our results support a simple stress hypothesis because the negative effects of malnutrition accumulated over successive generations. Density-dependent deterioration in plant quality is thought to be an important factor governing the population dynamics of outbreaking insects and we hypothesize that chronic nutritional stress can be a driver of outbreak declines of C. fumiferana, and of forest insects in general.