Pine sawfly defence and variation in host plant resin acids: a trade-off with growth

Abstract.

• 1 The defence capability of pine sawfly larvae reared on Scots pines with low or high concentrations of resin acids was determined in a field experiment, where larvae were exposed to predatory ants, and in a laboratory experiment in which the rate of defence droplet replenishment was measured.
• 2 Larvae on low resin-acid shoots disappeared faster than larvae on high resin-acid shoots when exposed to predatory ants.
• 3 Larvae fed high resin-acid needles produced 50% larger defence droplets than larvae fed low resin-acid needles.
• 4 In an additional experiment high predation pressure was simulated by repeatedly removing defence droplets. Larvae responded by feeding more on tissue rich in resin acids, but at the cost of a reduction in growth rate.
• 5 Based on these data and earlier findings that survival and development of sawfly larvae are negatively affected by high resin-acid concentrations, we conclude that pine sawflies face a dilemma of conflicting demands, i.e. although high needle resin-acid concentrations enhance defence capability, they also reduce growth and survival. Our data also suggest that it is not necessarily the ovipositing female that has to balance this trade-off: plasticity in larval feeding behaviour provides larvae with a means to at least partly solve the dilemma.

     

Variability of Norway spruce (Picea abies L.) needles; performance of spruce sawflies (Gilpinia hercyniae Htg.)

Summary Larvae of the spruce sawfly Gilpinia hercyniae were reared on whole branches of Norway spruce, Picea abies. Top and low branches were selected from flowering and nonflowring stands. Sawfly performance from the larval L₂-stage until the fertile egg stage of the next generation was recorded. Growth and development were best on top branches from the flowering stand, poorest on branches from windblown, partly derooted and heavily flowering trees. Analysis of some 30 biochemicals in needles and faeces was performed. New needles had the highest concentrations of some nutrients (total nitrogen, amino acids), however, G. hercyniae larvae only fed on old needles, a 100% mortality being recorded on newly flushed needles, probably because these needles also contained the highest concentrations of the secondary compound, quinic acid. Old needles showed high variability in concentrations of nutrients and secondary compounds. Regression analysis demonstrated that the total amount of carbohydrates (glucose, fructose and sucrose) was significantly correlated with the larval linear growth rate, the maximal larval weight, the cocoon weight and the number of eggs per female. In these performance variables 72–88% of the variation could be explained by five biochemicals. Survival rates of larvae or pupae and the instantaneous growth rate could not be explained by the biochemical variables. Only weak correlations were found between nitrogen or amino acids and performance variables. Assimilation of the various biochemicals was calculated and showed high rates (90–97%) in hexoses and lower rates (38–65%) in total nitrogen and amino acids.     

Host plant influence on chemical defense in conifer sawflies (Hymenoptera: Diprionidae)

Host diet affects the defensive efficacy of Neodiprion sawflies. In laboratory assays with wood ants (Formica obscuripes), secretions from larvae reared on Pinus banksiana were the most repellent, while those from P. resinosa feeders were the least so. This was explained diterpene resin acid content, but not total monoterpene content. The terpene content of regurgitant generally reflected dietary concentrations. Compounds were sequestered nonselectively by larvae. Host-based differences in defense persisted at the behavioral level. P. banksiana feeders regurgitated greater volumes of fluid and were less likely to be disabled or killed by ants in one-on-one interactions than were larvae fed on P. resinosa. The defensive advantages of host diet conflicted with developmental requirements. N. sertifer reared on P. banksiana (the best diet for defense in all cases) had lower cocoon weights (a correlate of fecundity) than those reared on other diets, and had prolonged larval development compared to insects fed P. sylvestris. No such tradeoff was detected in N. lecontei. Larvae of both species strongly preferred P. banksiana over P. resinosa in feeding choice assays.     

Neodiprion sertifer defoliation causes long-term systemic changes of oxidative enzyme activities in Scots pine needles

The aim of the experiments reported here was to study possible long-term effects of Neodiprion sertifer Geoff. (Hymenoptera: Diprionidae) herbivory, or artificial defoliation, on oxidative enzyme activities in Scots pine (Pinus sylvestris L.) needles as a consequence of induced defense responses. During year 1 (the first season), defoliation by N. sertifer, which feeds on previous season’s needles, did not result in statistically significant changes in polyphenol oxidase activity in the current year’s needles. In contrast, defoliation did lead to increased peroxidase activity in those needles. In the second season (year 2) N. sertifer defoliation of pine seedlings, also defoliated in the previous season either by larvae or artificially, resulted in a decrease of peroxidase activity in the current year’s needles. No significant differences between treatments carried out in year 1 were found in year 2 for peroxidase activity in the previous year’s needles. However, defoliation in year 1 by N. sertifer resulted in decreased needle consumption, and higher mortality of larvae, in year two. These results indicate the existence of long-term changes in needle oxidative enzyme activities as a consequence of N. sertifer feeding.     

Influence of plant quality on pine sawfly population dynamics

The contribution of plant quality to the population dynamics of herbivorous insects has been an issue of much controversy. Many studies have documented how variable plant quality differentially influences the survival and fecundity of insect individuals. Whether or not such effects can be translated to the level of insect populations is, however, not clear. In order to test this hypothesis one needs to combine processes at both the level of the individual and the population. This is difficult with an empirical approach, but could be achieved by means of modeling given that appropriate data exist for both levels of organization. In this paper we report on a model developed to analyze whether altered Scots pine (Pinus sylvestris) quality can contribute to the build‐up of populations of the European pine sawfly (Neodiprion sertifer). Experimental data on responses of sawfly larvae to variable plant quality, i.e. needle concentrations of resin acids, were used to parameterize the model. Larval survival and sawfly fecundity are reduced at high resin acid concentrations. However, high resin acid concentrations are, at the same time, beneficial because larval defense against predators is enhanced. In the model, data on individual responses were combined with literature data at the population level; a type III functional response related to cocoon predation was presumed to be the density‐dependent process regulating sawfly populations. The analysis showed that the risk for an outbreak is high when needle resin acid concentration (r) or larval predation pressure (p) is low. When r or p is high there is no risk. By analyzing different scenarios it was found that small changes in r and p can result in the sawfly population moving from low to high outbreak risk. Changes of the same, or larger, magnitude in r have been observed in empirical studies. The role of tritrophic interactions was also considered. This was done by removing the positive effects of resin acids on larval performance in the model. It was found that the anti‐predator defense of N.sertifer makes it prone to outbreak under wider combinations of r and p than an insect without the defense. We conclude that small changes in a density‐independent factor, such as needle chemistry, can have significant effects on herbivore population dynamics because increased fecundity and survival caused by needle quality may allow the population to escape the control of density‐dependent factors, such as cocoon predation.     

Laboratory studies on the larval food habits of the ground beetle Amara (Curtonotus) gigantea (Motschulsky) (Coleoptera: Carabidae: Zabrini)

Ground beetles (Coleoptera: Carabidae) are important components of soil ecosystems in temperate zones, but our knowledge regarding the ecology of many species is limited. This study examined larval food habits in the ground beetle Amara (Curtonotus) gigantea (Motschulsky) using laboratory‐rearing experiments. Because this beetle is a member of the tribe Zabrini, which includes several species that exhibit granivory during larval and/or adult stages, three diet types were tested, with starvation as a control: Tenebrio larvae, mixed seeds (Bidens pilosa (Asteraceae), Setaria spp., Digitaria ciliaris (both Poaceae), and Humulus scandens (Moraceae)), and Tenebrio larvae + mixed seeds. Because of high larval mortality during overwintering under laboratory‐rearing conditions, survival and the duration of development through pre‐overwintering stages (first and second instars) were compared. Larvae fed the diet of Tenebrio larvae + mixed seeds showed the highest survival (89%), followed by the diet of Tenebrio larvae (83%), but the difference between the two treatments was not significant; similarly, developmental durations did not differ between these two diets. Larvae fed the mixed‐seeds diet showed markedly lower survival, and no larvae reached the second instar. Therefore, for A. gigantea larvae, animal food results in high larval performance, whereas seeds provide a low value diet; animal food is essential for larval development, but seeds provide no benefit, even as a dietary supplement. Overall, the present results suggest that A. gigantea larvae are omnivorous but with a mainly carnivorous food habit.     

Performance of pine looper Bupalus piniarius larvae under population build-up conditions

In the first year of an outbreak, Bupalus piniarius larvae, encounter intense crowding. In the later stages of larval development, they are forced to feed on the non-preferred current-year needles of Scots pine or even on alternative hosts. It was hypothesized that larval feeding on a non-preferred resource (current-year needles) will negatively affect B. piniarius performance. It was also hypothesized that larval mutual interference (crowding without competition for food) will have negative additive effects. These hypotheses were tested in laboratory and field experiments. Fourth instar larvae were reared singly and under crowded conditions in cohorts of ten. Larvae in both situations were reared on control branches (containing both mature and current-year shoots) and branches containing only current-year shoots. Crowded larvae were reared also on Norway spruce, an alternative host. Crowding and feeding on a non-preferred resource had opposite effects on B. piniarius larval performance. Crowding in the late larval instars enhanced larval performance while absence of the preferred resource had negative effects. Larval growth rate was higher and development time was shorter for larvae exposed to crowded conditions than for solitary larvae. There was, however, no difference between the groups in final pupal weights or survival. Survival was 25% lower for larvae feeding on non-preferred current-year needles and pupal weights 9% lower, compared with results for larvae feeding on mature needles. Larvae feeding on Norway spruce suffered greatly extended development time, 82% lower survival, and resulted in 60% lighter pupae compared with conspecifics on Scots pine. It was concluded that not only quantity but also quality of the available food resource is critical for B. piniarius development.     

The dependence of development and fecundity of Samea multiplicalis on early larval nitrogen intake

Larvae of the moth Samea multiplicalis (Lepidoptera: Pyralidae), developed more rapidly when their food plant, Salvinia molesta was richer in nitrogen. Larvae that fed on plants with less than 1.35% nitrogen during their first instar completed the larval stage in 18.0–24.2 days, after passing through six instars. In contrast, larvae that had fed on food richer in nitrogen during their first instar completed the larval stage in 14.1–17.4 days, and passed through five instars. Experience of nitrogen in later instars had little additional effect on total larval development time. Oöcyte complements of 1-day old adult females was correlated with the mean food nitrogen content over the larval period. Food nitrogen content experienced by larvae in earlier instars was as important as that in the final instars, in determining number of mature oöcytes, which suggests early conditioning of the nitrogen use effciencies of later instars.     

Larval mortality and development of Pseudoplusia includens (Lepidoptera: Noctuidae) reared on a transgenic Bacillus thuringiensis-cotton cultivar expressing CryIAc insecticidal protein.

The effects of a transgenic Bacillus thuringiensis (Bt)-cotton cultivar (DPL 32) on three instars of the soybean looper, Pseudoplusia includens (Walker), were determined in laboratory studies. First, third, and fifth instars were fed field collected Bt-cotton leaves for 1, 2, four and 7 d or until pupation, and then transferred to artificial diet. Mortality during the larval stage increased linearly in response to an increase in the length of feeding time on Bt-cotton by first and third instars. The maximum mortality of about two out of three larvae occurred for first instars fed on Bt-cotton until pupation. For the fifth instar, there was no significant response to feeding time; however, most of these larvae reached pupation before 4 d of feeding on Bt-cotton. The length of the larval developmental period also increased linearly with an increase in feeding time on Bt-cotton in first and third instars; again, there was no significant response in the fifth instars. For both mortality and larval developmental time, the linear trend lines for the first and third instars were quite similar. Pupal weight declined linearly in the first and fifth instars in response to feeding time on Bt-cotton. Although pupal weight also declined for third instars, the response was not linear. The effect of Bt-cotton appears not to extend past pupation in that there were no significant responses in mortality and developmental time of pupae during the pupal stage. These data indicate that larvae surviving Bt-cotton are adversely affected in several ways, which should be considered in evaluating Bt-cotton suppression of soybean looper infestations.     

Fish-induced variation in abdominal spine length of Leucorrhinia dubia (Odonata) larvae?

The lengths of dorsal and lateral spines on abdominal segments 4, 6 and 9 were measured on last-instar larvae of Leucorrhinia dubia (Odonata) from seven natural lentic systems containing fish and nine systems lacking fish. Larvae from systems with fish had significantly longer spines than larvae from systems without fish. In contrast, lake/pond area and pH had no effect on the length of spines. The length of the spines was not correlated with larval size, but there was a high correlation between the length of the three spines measured. Also, abundances of L. dubia larvae differed between systems, being significantly lower in systems with fish. Laboratory experiments showed that perch (Perca fluviatilis) handled long-spined larvae for significantly longer times than short-spined larvae. The results suggest that fish predators may induce morphological defences in insects.     

Effect of dietary selenium supplementation on resistance to baculovirus infection

We have examined the effects of dietary selenium (Se) supplementation on larval growth and immunocompetence of the lepidopteran pest, the cabbage looper, Trichoplusia ni. Supplementation of the diet of T. ni larvae with 10–20 ppm Se resulted in a 1 day delay in pupation. The effects of the addition and/or removal of dietary Se on total Se bioaccumulation and sequestration were determined by neutron activation analysis of pupae. Early penultimate instar larvae moved from selenium containing diet to basal diet lost total pupal Se content down to the level of those fed basal diet. Conversely, larvae moved from basal diet to diet containing additional Se rapidly attained pupal Se levels comparable to larvae fed Se throughout larval development. Therefore, dietary Se is rapidly accumulated or lost during larval development, but significant amounts are sequestered from diet into pupae. Larvae were reared on diet supplemented with 5 or 10 ppm Se until the onset of the penultimate instar then infected per os with increasing concentrations of the fatal baculovirus Autographa californica multiple nucleopolyhedrovirus (AcMNPV). Larvae fed Se in the penultimate and ultimate instars were more resistant to viral infection than larvae not fed Se in the final instars. This study indicates that dietary Se levels rapidly impact Se assimilation and sequestration and that tissue Se levels are an important factor in resistance to AcMNPV infection in larval T. ni.     

The effect of prey density on the developmental time of larvae of an aquatic beetle: <Emphasis Type="Italic">Tropisternus setiger</Emphasis> (Insecta,Coleoptera: Hydrophilidae)

Predaceous larvae of the water scavenger beetle Tropisternus setiger (Germar) are common inhabitants of variable environments in which prey availability may vary widely. We conducted laboratory experiments to assess the effect of prey density on developmental times and survivorship of the preimaginal stages of T. setiger. We also examined the effect of the number of consumed prey on the larval size of instar III. Four different prey densities (one, two, four, and eight preys a day) were tested and both developmental time and survivorship differed significantly among them. Larvae fed one or two preys daily showed a longer developmental time and a lower survivorship than larvae fed four or eight preys a day. Moreover the consumption of four preys a day increased larval developmental success, and to consume one prey a day affected survivorship through the larval period. On the other hand, prey density had no effect on the final larval size. Handling editor: K. Martens     

Effects of group size and pine defence chemicals on Diprionid sawfly survival against ant predation

The defence chemicals and behavioural adaptations (gregariousness and active defensive behaviour) of pine sawfly larvae may be effective against ant predation. However, previous studies have tested their defences against very few species of ants, and few experiments have explored ant predation in nature. We studied how larval group size (groups of 5 and 20 in Neodiprion sertifer and 10, 20 and 40 in Diprion pini) and variation in levels of defence chemicals in the host tree (Scots pine, Pinus sylvestris) affect the survival of sawfly larvae. Food preference experiments showed that ants do eat sawfly larvae, although they are not their most preferred food item. According to our results, ant predation significantly increases the mortality rate of sawfly larvae. Larval mortality was minor on pine tree branches where ant traffic was excluded. We also found that a high resin acid concentration in the host tree significantly decreased the mortality of D. pini larvae when ants were present. However, there was no such relationship between the chemical concentrations of the host tree and larval mortality for N. sertifer. Surprisingly, grouping did not help sawfly larvae against ant predation. Mortality risk was the same for all group sizes. The results of the study seemingly contradict previous understanding of the effectiveness of defence mechanisms of pine sawfly against ant predation, and suggest that ants (Formica exsecta in particular) are effective predators of sawfly larvae.An erratum to this article can be found at     

Simulated acid rain reduces the susceptibility of the European pine sawfly (Neodiprion sertifer) to its nuclear polyhedrosis virus

Summary The study dealt with the effect of simulated acid rain (both H₂SO₄ and HNO₃; acidities of pH 4 and pH 3) on the susceptibility of the larvae of Neodiprion sertifer to its nuclear polyhedrosis virus. Scots pines growing in a subarctic area with low ambient pollution levels were irrigated with simulated acid rain during two summers. Neodiprion larvae fed with foliage from the experimental trees were infected with a dilute virus suspension. The acid treatment of host trees had a significant effect on the proportion of virus-treated larvae alive 16 days after the virus application: there were almost no differences between the controls and the pH 4 irrigation group, but on the needles of pH 3-treated trees larval survival was twice as high as with other treatments. The direct spraying of acid water on the needles before they were fed to the larvae did not significantly affect the survival of virus infected larvae. Our results suggest that acid rain may reduce the susceptibility of Neodiprion larvae to virus disease via changes in the quality of pine foliage.     

Disengtangling the evolution of weak warning signals: high detection risk and low production costs of chemical defences in gregarious pine sawfly larvae

Evolution of costly secondary defences for a cryptic prey is puzzling, if the prey is already well protected by camouflage. However, if the chemical defence is not sufficient to deter all predators, selection can favour low signal intensity in defended prey. Alternatively, if the costs of chemical defence are low or cost-free, chemical defences can be expected to evolve also for non-signalling prey, particularly if conspicuous signalling is costly. We tested these assumptions with pine sawfly larvae (Neodiprion sertifer and Diprion pini) that are cryptically coloured and chemically defended with resin acids sequestered from their host plant (Pinus sp.). Larvae feed in large aggregations, which we hypothesise could function as a signal of unprofitability. Our results show that even though the birds found N. sertifer larvae unprofitable in the controlled laboratory assays, they continued attacking and consuming them in the wild. When we tested the signal value of aggregation we found that a large group size did not offer protection for a defended larva: the survival was higher in groups of 10 individuals compared to groups of 50, suggesting increased detectability costs for individuals in larger groups. Finally, we tested how costly the production and maintenance of a chemical defence is for D. pini larvae by manipulating the resin acid content of the diet. We did not find any life history or immunological costs of the chemical defence for the larvae. In contrast, pupal weights were higher on the high resin diet than on the low resin diet. Also, larvae were able to produce higher amounts of defence fluids on the high diet than on the low diet. Thus, our result suggests high detectability costs and low production costs of defences could explain why some unprofitable species have not evolved conspicuous signals.     

Food availability controls the onset of metamorphosis in the dung beetle Onthophagus taurus (Coleoptera: Scarabaeidae)

In nature, larvae of the dung beetle Onthophagus taurus (Schreber 1759) are confronted with significant variation in the availability of food without the option of locating new resources. Here we explore how variation in feeding conditions during the final larval instar affects larval growth and the timing of pupation. We found that larvae respond to food deprivation with a reduction in the length of the instar and premature pupation, leading to the early eclosion of a small adult. To achieve pupation, larvae required access to food for at least the first 5 days of the final instar (= 30% of mean third‐instar duration in control individuals), and had to exceed a weight of 0.08 g (= 58% of mean peak weight in control individuals). Larvae that were allowed to feed longer exhibited higher pupation success, but increased larval weight at the time of food deprivation did not result in increased pupation success except for larvae weighing > 0.14 g. Larvae responded to food deprivation by initiating and undergoing the same sequence of developmental events, requiring the same amount of time, as ad libitum‐fed larvae once those had reached their natural peak weight. Our results reveal a striking degree of flexibility in the dynamics and timing of larval development in O. taurus. They also suggest that premature exhaustion of a larva's food supply can serve as a cue for the initiation of metamorphosis. Premature metamorphosis in response to food deprivation has been documented in amphibians, but this is, to the best of our knowledge, the first time such a behaviour has been documented for a holometabolous insect. We discuss our findings in the context of the natural history and behavioural ecology of onthophagine beetles.     

Larval mortality of pine sawflies [Hymenoptera: Diprionidae] in relation to pollution level: A field experiment

Larval mortality ofNeodiprion sertifer (Geoffroy),Diprion pini (L.) andGilpinia pallida (Klug) were studied in field experiments around a factory complex in southwestern Finland. Larval colonies were transferred on the shoots of Scots pines growing at different distances from the emission source. Larval mortality was highest near the factories. InN. sertifer, larval mortality caused by the nuclear polyhedrosis virus was higher and cocoon mortality caused by parasitoids was lower near the pollutant source. The most abundant parasitoid species wereSynomelix scutulata (Hartig) andLamachus eques (Hartig). 16–67% of theN. sertifer, 0–5% of theD. pini and the 73–100% ofG. pallida cocoons contained parasitoids oviposited during the larval period of the sawflies.      

马尾松毛虫对马尾松和湿地松的选择与适应研究

用马尾松老叶、新叶,湿地松老叶、新叶,当年生马尾松新叶、湿地松新叶和当年生以20％和40％β-蒎烯处理过的马尾松新叶作食料饲养马尾松毛虫。结果表明：马尾松毛虫幼虫在第1代发生期取食马尾松老叶、马尾松新叶、湿地松老叶和湿地松新叶等4种不同质量的食料后,因其食料中10种必需氨基酸含量和挥发性物质β-蒎烯含量的不同,导致幼虫的存活率差异显著。其中,取食马尾松老叶的幼虫成活率最高（44％）,依次为马尾松新叶（32.7％）、湿地松老叶（4％）,而取食湿地松新叶的幼虫全部死亡。通过马尾松针叶增加外援β-蒎烯喂养马尾松毛虫实验,表明增加了外源β-蒎烯的松针饲养的马尾松毛虫幼虫与用同样松针未增加外源β-蒎烯饲养的幼虫相比,死亡率明显提高;松树中β-蒎烯含量的增加与幼虫成活率呈显著负相关。由此,我们阐明了马尾松与湿地松对马尾松毛虫的抗虫机制。     

Influence of food and larval age on the defensive chemistry of Saturnia pyri

Scolus secretions and hemolymph of caterpillars of Saturnia pyri fed with two different foodplants (Crataegis monogyna, Prunus spinosa) were chemically analyzed and their chemical similarities determined. The secondary-compound patterns obtained for the two body fluids showed no significant differences when compared between the two groups of alternatively fed last-instar larvae. Thus, the composition of these fluids of full-grown caterpillars is not influenced by the larval diet. However, younger larvae on P. spinosa revealed a diversity of compounds differing significantly from that of larger caterpillars fed with either C. monogyna (both body fluids) or P. spinosa (hemolymph only). This indicates that, on the one hand, the hemolymph composition is adapted to the changing physiological requirements of the given instars whereas, on the other hand, the defensive mixtures remain unaltered in the late larval instars due to a constant spectrum of potential enemies.     

Developmental plasticity in larval development in the echinometrid sea urchin Evechinus chloroticus with varying food ration

Developmental plasticity in the morphology of planktotrophic echinoid larvae has been studied primarily in temperate sea urchins from the Northern Hemisphere. These studies have shown that echinoplutei with reduced food ration generally respond by increasing the length of the larval arms, and thus the length of the ciliated band, and delaying formation of the rudiment. Using the endemic New Zealand sea urchin Evechinus chloroticus we tested for the presence of developmental plasticity in larvae fed a high or low algal food ration (6000 or 600 Dunaliella cells/ml), or with no algal food. Our results show that developmental plasticity in larval form is seen in a Southern Hemisphere representative of the Family Echinometridae, but the general pattern of longer arms with low food ration was only seen in the earliest part of development. Larvae in the High food treatment were largest in all dimensions and formed rudiments within 23 days of fertilization. Larvae fed a low algal ration or no algal food stalled at the four-arm echinopluteus stage, and were significantly smaller in size, and differed in shape, when compared in a multivariate analysis to the High food treatment. We suggest that the response of echinoplutei to low levels of particulate food is a species-specific trait, depending in part on the level of dependence of the larvae on exogenous food. Previous studies in Lytechinus variegatus, and the present study with E. chloroticus, suggest that in species where development stalls at the four-arm stage, the pattern of longer larval arms with low food ration will only be seen during the initial early period of larval growth. Additionally our results show that there can be significant variation in larval morphology between replicate jars in the same feeding treatment, suggesting that future research on developmental plasticity should also consider differences in larval morphology between culture containers.