The genus Leptopilina comprises species of parasitoid wasps that primarily attack early instar larvae of drosophilid species, some of which are well-known agricultural or nuisance pests. To unravel host-parasitoid interactions and to use parasitoids effectively in biological control, the correct taxonomic assignment is of crucial importance. In the course of the German Barcode of Life III: Dark Taxa (GBOL III: Dark Taxa) project, we assembled specimens of all seven species of Leptopilina of the Western Palearctic. Using CO1 barcode sequence analyses and morphological examination allows us to evaluate species limits in an integrative approach, to provide extended species diagnoses as well as an updated key to the species of the Western Palearctic. This work also summarizes current knowledge on Western Palearctic Leptopilina species, including their distribution, population parameters, habitats, flight periods, and hosts. We provide the first country records of three Leptopilina species for Belgium, including the non-native L. japonica, propose two species level synonymies and move one Western Palearctic species out of Leptopilina. Finally, to facilitate taxonomic research on Leptopilina in lesser-known regions, we provide a list of non-Western Palearctic species, including the moving of 13 species into Leptopilina. These changes also result in two new generic synonymies. In conclusion, Western Palearctic Leptopilina can be clearly diagnosed both morphologically and molecularly. Comparable ease of identification and cultivation, as well as the economic interest in potent biological control agents against drosophilid pests make them ideal organisms for basic and applied research.

Damage to planted conifer seedlings caused by the pine weevil Hylobius abietis (L.) is a severe and persistent threat to successful forest regeneration in Europe. Various countermeasures are available, which vary in effectiveness, costs and environmental impact, but none are ideal for all situations. Therefore, there is strong interest in robust assessments of damage risks, as they would enable more cost-effective and environmentally friendly forest pest management. It has been suggested that numbers of adult pine weevils caught in host-odour baited traps placed in regeneration sites may be valuable in such risk assessments. However, published studies provide at most weak support for the hypothesis that trap catch data provide adequate predictions of damage. Therefore, we conducted a two-year field study, designed to determine the relationship between weevil trap catch and subsequent damage at 10 regeneration sites in central Sweden. Site factors that might influence pine weevil feeding on seedlings were recorded and used as explanatory variables in the analysis. Stoniness was the only site factor identified as having a significant effect; damage mainly increased with increases in stoniness. No significant correlation was detected between damage to planted conifer seedlings and numbers of pine weevils trapped in the same locations. We suggest that this lack of correlation between weevil numbers and damage is due to planted seedlings only constituting a minor part of the weevils food intake and considerable between-site variation in availability of food sources other than seedlings. Therefore, assessment of pine weevil numbers appears unlikely to be useful for predicting damage risk at specific regeneration sites.

To understand colonization success of an invasive species we need to know the origin of the founders, where and when they were introduced, and how they spread from the introduction site(s) through the landscape. Admixture of different genetic lineages from multiple introductions is generally hypothesized to be beneficial to invasive species thanks to adaptive variation and heterozygosity-fitness correlations. In this study, population genetic and landscape data was gathered for Roesel’s bush-cricket, Roeseliana roeselii a small bush-cricket common in central and eastern Europe that currently is expanding its range in northern Europe. We examined how colonization history and landscape structure affect the spread of the species and its population genetic structure, as a consequence of multiple introductions. Using comprehensive information of the species ecology and dispersal, together with genetic structure inferred from samples from 29 locations in central Sweden (we employed data published by Preuss et al., 2015), we found that two parapatric founding lineages have coexisted with very little gene flow during a long time span. An isolation-by-distance pattern and a decrease of genetic diversity toward marginal areas were more pronounced in the lineage situated in forest dominated landscapes. Our findings are in strong contrast to the hypothesis that different genetic lineages will admix when introduced to the same area. The presence of the separate lineages decades after introduction and without physical barriers for gene flow shows that some mechanism prevents them from admixture. One possibility is that the lineages with different genetic setups have adapted independently to local conditions and their admixture resulted in loss of locally adapted genotypes and hybrid offspring, less viable than the respective ancestral genotypes. However, an alternative post-mating reproductive barrier and hybrid breakdown phenomenon should also be considered. Our data indicate that besides landscape characteristics, human transportation of agricultural goods may play an important role for the overall spatial genetic pattern of the species in the study area by aiding the spread of the species.

Forest regeneration can be compromised by insect damage to newly-planted conifer seedlings, with the pine weevil, Hylobius abietis, being the most economically important pest in Europe. Seedling protection strategies include physical barriers, silvicultural measures and insecticides, while the potential benefits of Associational Effects (AE) have been little explored. Associational Resistance (AR) or Susceptibility (AS) arise when neighbouring plants decrease or increase, respectively, the likelihood and extent of attack on a focal plant. We investigated the potential of European beech, Fagus sylvatica, to mediate AE for Norway spruce, Picea abies, seedlings against pine weevil damage. First, we examined the effects of neighbor identity on damage to P. abies at a small scale, using choice arenas in the lab. Then, in the field, we examined these effects at a larger scale using plots containing only Norway spruce, or both species. We found that P. abies seedlings were attacked and damaged less by weevils when beech was their close neighbor, relative to having another Norway spruce as a neighbor in the lab. Yet, no difference in damage between only spruce and mixed seedling plots was found in the field. Our results indicate that the susceptibility of P. abies to H. abietis can be influenced by neighbor identity, and effects can vary with inter-plant distance. In close proximity, the presence of the non-host F. sylvatica can alter pine weevil feeding behaviour and thus, has the potential to mediate AR. However, these associational effects appear not to provide enhanced seedling protection at a larger scale.

A plant's induced defense system can be triggered by the application of the plant hormone methyl jasmonate (MeJA), and recent research suggest that MeJA treatment may become a tool for protection of conifer seedlings against insect herbivory (e.g. by the pine weevil Hylobius abietis). A side-effect of MeJA application is temporarily reduced height growth. This has generally been considered as negative, but in forest tree nurseries this could instead be beneficial since it is commonly desired to stop the growth of nursery seedlings in late summer. Artificially longer dark periods (long nights/short days) are widely used in high-latitude nurseries to terminate height growth and induce freezing tolerance. However, long night treatment requires specialized nursery equipment and are labor intensive. Therefore alternatives are sought after. We compared long-night and MeJA treatments by following the growth of Norway spruce (Picea abies) seedlings throughout one season. The regulatory effect of MeJA on height growth was similar if not even better than that of long nights, i.e. it was terminated faster. However, MeJA treatment also reduced root growth and delayed the development of freezing tolerance. MeJA may therefore not replace long-night treatments, but it could facilitate a more flexible application of long nights by gaining a longer time interval during which this treatment can be used without risking the seedlings growing too large.

The pine weevil Hylobius abietis is an important pest causing severe damage to conifer seedlings in reforestation areas in Europe and Asia. Plants that have no evolutionary history with the pine weevil are of special interest in the search for compounds with a strong antifeedant activity. Thus, the essential oils of nine aromatic plants, viz Amomum subulatum, Cinnamomum tamala, Curcuma longa, Laurus nobilis, Ocimum basilicum, Origanum majorana, Origanum vulgare, Syzygium aromaticum and Trachyspermum ammi were extracted by hydrodistillation. The essential oil constituents were identified by gas chromatography-mass spectrometry, and antifeedant properties towards the pine weevil were assessed using choice feeding bioassay. The essential oils of C. longa, O. majorana, S. aromaticum and T. ammi showed an excellent antifeedant activity towards the pine weevil for 24 hr, whereas the essential oil of other plants showed the activity for 6 hr. There was a positive correlation between the amount of benzenoid compounds and the antifeedant activity of the essential oils. This study suggests that pine weevil non-host plant compounds have potential to be used for the protection of seedlings against pine weevil feeding. However, further study will be needed to explore the antifeedant activity of individual components and oils in the laboratory as well as in the field.

BACKGROUND:The pine weevil (Hylobius abietis) is a major forest regeneration pest causing high levels of seedling mortality and economic losses. Current management relies on silviculture, stem coatings and insecticides. Here we evaluated for the first time the effects of Bacillus thuringiensis (Bt) strains on H. abietis adults: two producing the Coleoptera-targeted toxins Cry3Aa (Bt tenebrionis NB-176) and Cry8Da (Bt galleriae SDS-502), and one producing the Diptera-targeted Cry10A (Bt israelensis AM65-52). Choice and nonchoice assays using individual and mixtures of Bt formulations, containing these strains respectively, were conducted. RESULTS:We found that Bt had toxic and lethal effects on H. abietis, but effects varied with strain and formulation concentration. The Diptera-targeted Bt israelensis had the most negative effects on weevil weight, feeding and mortality (70-82% feeding reduction, 65-82% greater mortality than control), whereas the effect was lower for the Coleoptera-specific Bt tenebrionis (38-42%; 37-42%) and Bt galleriae (11-30%; 15-32%). Reduced weevil feeding was observed after 3 days, and the highest mortality occurred 7-14 days following Bt exposure. However, we found no synergistic toxic effects, and no formulation combination was better than Bt israelensis alone at reducing consumption and survival. Also, pine weevils were not deterred by Bt, feeding equally on Bt-treated and non-Bt treated food. CONCLUSION:There is potential to develop forest pest management measures against H. abietis that include Bt, but only the Diptera-targeted Bt israelensis would provide effective seedling protection. Its Diptera-specificity may need reconsideration, and evaluation of other Bt strains/toxins against H. abietis would be of interest. © 2020 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

The pine weevil (Hylobius abietis) is one of the most devastating pests of regenerated coniferous forests in northern and central Europe. Although it is also present and potentially harmful in southern Europe, little attention has been paid to this pest in that region. With the aim of quantifying the potential risk of the pine weevil as a forest pest in southern Europe, we performed an intensive assessment of weevil abundance and damage in 21 Pinus pinaster and P. radiata plantations established in recent coniferous clear-cuts in Galicia (NW Spain), i.e. in the south-western part of the natural distribution of H. abietis. The effect of several silvicultural and landscape factors as modulators of weevil risk was analyzed. Abundance of adult pine weevils was highly variable across the sampling plots, with the number of trapped insects significantly correlated with mean air temperature during each trapping period. Damage rate was, however, fairly constant across the whole study period. No significant relationship between pine weevil abundance and damage was observed. Up to 85% of the planted seedlings were attacked and the mortality rates reached 45% in some plots. No weevil preferences were found between the two pine species. Pine weevil damage was positively related to the amount of slash (logging waste) on the ground and negatively related to the time since clear-felling occurred. Once these factors were accounted for, the abundance of mature coniferous forest in the vicinity (500-2000 m) of the clear-cut significantly and positively affected pine weevil damage. We conclude that a high risk of pine weevil damage does exist in NW Spain. Low-intensity forest management together with the scarcity and discontinuity of mature conifer forest in the region seem to be the reason why this forest pest is largely ignored by forest owners and forest-health services. If the forest sector intensifies in this area, care should be taken to protect pine plantations against this devastating pest.

This study concludes an extensive investigation of antifeedants for the pine weevil, Hylobius abietis (Coleoptera: Curculionidae), an economically important pest of planted conifer seedlings. Building on the previously reported antifeedant effects of benzoates and phenylpropanoids (aromatic compounds with one- or three-carbon-atom substituents on the benzene ring), we here report the antifeedant effects of compounds with two-carbon-atom side chains (i.e., phenylacetates). We also present new results; the best antifeedants from the benzoate class were tested at 10-fold lower concentrations in order to find the optimal antifeedants. Generally, for all three compound classes, efficient antifeedants were found to have one or two methyl, chloro, or methoxy substituents on the aromatic ring. For monosubstituted phenylpropanoids, the substituent preferably should be in the para-position. In the search for synergistic antifeedant effects among the three compound classes, combinations of compounds from the three classes were tested in binary and ternary mixtures.

Somatic embryogenesis (SE), a clonal propagation method utilizing somatic cells, occurs under conditions that activate plant stress adaptation mechanisms such as production of protective secondary metabolites. Surprisingly, possible differences in susceptibility to insect pests between SE-generated and conventionally cultivated plants have not been previously explored. Here, we recorded frequencies and levels of bark-feeding damage by pine weevils (Hylobius abietis) in two large field trials, consisting of emblings (SE-propagated plants) and seedlings from 50 half-sib Norway spruce (Picea abies) families. We found that emblings were less frequently attacked by pine weevils, and when attacked, they were damaged to a lesser extent than seedlings. Moreover, we detected significant additive genetic variation in damage levels received by plants, indicating a heritable component to differences in resistance to insect herbivory among half-sib families. We present first-time evidence that emblings can be more resistant than seedlings to herbivorous insect damage, thus, SE appears to confer a previously unknown plant protection advantage. This finding indicates novel avenues to explore mechanisms underlying plant resistance and new approaches to develop non-toxic measures against insect pests.

Adult pine weevils (Hylobius abietis (L.)) feed on the tender bark of branches and roots of mature conifer trees and on the stem bark of conifer seedlings. Their feeding on mature trees does not cause any economic damage, but their feeding on planted seedlings is so devastating that the pine weevil is considered one of the most important forest pest insects in Europe. We asked whether the pine weevil prefers seedlings over other regularly utilized food sources. This question is of particular interest because new approaches to seedling protection are based on decreasing any preference for seedlings by using less palatable plants or by enhancing their defence (by genetic selection or by methyl jasmonate treatment). In a laboratory choice experiment we tested pine weevil feeding preferences for seedlings compared with branches and roots from mature trees (separately for Norway spruce and Scots pine). Pine weevils preferred roots, but not branches, of Norway spruce over seedlings of the same species. With Scots pine there were no clear preferences, but the weevils showed a tendency to prefer roots over seedlings. These results provide support for seedling protection approaches that attempt to redirect pine feeding from planted seedlings to other food sources.

Larvae of Strobilomyia flies (Diptera: Anthomyiidae) are serious pests in conifer-seed orchards because they feed on the seed inside the cones. Figitid parasitoids (Hymenoptera: Cynipoidea) of Strobilomyia larvae in conifer cones are commonly reported but under various generic names. It is argued here that, across the entire Holarctic region, these figitids belong to Amphithectus and perhaps also to Sarothrus (Figitinae), but not to Melanips (Aspicerinae), contrary to some reports. We conclude that the identity of the commonly found figitid associated with conifer cones (Larix and Picea) in Europe and Asia is Amphithectus austriacus (Tavares, 1928) comb. n. This is most likely considering the original description and the host association, although the type specimen of Seitneria austriaca Tavares, 1928 is lost. This species name takes priority over the recently described Amphithectus coriaceus Paretas-Martinez & Pujade-Villar, 2013. Seitneria Tavares, 1928 becomes a new junior synonym of Amphithectus Hartig, 1840, and Amphithectus coriaceus Paretas-Martinez & Pujade-Villar, 2013 becomes a new synonym of Amphithectus austriacus (Tavares, 1928) comb. n.

The pine weevil Hylobius abietis L. is an economically important pest insect that kills high proportions of conifer seedlings in reforestation areas. It is present in conifer forests all over Europe but weevil abundance and risk for damage varies considerably between areas. This study aimed to obtain a useful model for predicting damage risks by analyzing survey data from 292 regular forest plantations in northern Sweden. A model of pine weevil attack was constructed using various site characteristics, including both climatic factors and factors related to forest management activities. The optimal model was rather imprecise but showed that the risk of pine weevil attack can be predicted approximatively with three principal variables: 1) the proportion of seedlings expected to be planted in mineral soil rather than soil covered with duff and debris, 2) age of clear-cut at the time of planting, and 3) calculated temperature sum at the location. The model was constructed using long-run average temperature sums for epoch 2010, and so effects of climate change can be inferred from the model by adjustment to future epochs. Increased damage risks with a warmer climate are strongly indicated by the model. Effects of a warmer climate on the geographical distribution and abundance of the pine weevil are also discussed. The new tool to better estimate the risk of damage should provide a basis for foresters in their choice of countermeasures against pine weevil damage in northern Europe.

Replanting at appropriate times after harvesting a coniferous forest stand can help efforts to suppress seedling mortality caused by the pine weevil Hylobius abietis, but optimal times are uncertain. We hypothesized that planting in June rather than May in the third season after harvest would reduce feeding damage by the pine weevil and increase seedling survival rates in central Sweden, where new-generation weevils mainly fly away from their development sites in May/early June. An experimental test of the hypothesis in eight clear-cuts confirmed that planting in June instead of May reduced proportions of seedlings attacked by pine weevil, bark removal from seedlings' stems, and proportions of seedlings killed by feeding damage. These differences between seedlings planted in May and June declined to some extent with time but still remained significant after two growing seasons. The total seedling mortality after two seasons did, however, not differ significantly between seedlings planted in May and June. Overall, 29% of all seedlings were killed by pine weevil, 4.0% by Hylastes bark beetles, and 2.3% by drought. The results indicate that replanting in spring during the third season after harvest can advantageously continue until mid-June with respect to damage and mortality.

The pine weevil, Hylobius abietis, is a severe forest pest insect as it feeds on newly planted conifer seedlings. To identify and develop an antifeedant could be one step towards the protection of seedlings from feeding damage by the pine weevil. With the aim to trace the origin of the antifeedants previously found in feces of the pine weevil, we investigated the culturable bacteria associated with the gut and identified the volatiles they produced. Bacterial isolates were identified by 16S ribosomal RNA gene analysis. The volatile emissions of selected bacteria, cultivated on NB media or on the grated phloem of Scots pine twigs dispersed in water, were collected and analyzed by solid-phase microextraction gas chromatography-mass spectrometry. The bacterial isolates released a variety of compounds, among others 2-methoxyphenol, 2-phenylethanol, 3-methyl-1-butanol, 1-octen-3-ol, 3-octanone, dimethyl disulfide, and dimethyl trisulfide. A strong antifeedant effect was observed by 2-phenylethanol, which could thus be a good candidate for use to protect planted conifer seedlings against feeding damage caused by H. abietis.

Regeneration of northern conifer forests is commonly performed by reforestation with genetically improved materials obtained from long-term breeding programs focused on productivity and timber quality. Sanitary threats can, however, compromise the realization of the expected genetic gain. Including pest resistance traits in the breeding programs may contribute to a sustainable protection. Here we quantified the variation in different components of resistance of Norway spruce to its main pest, the pine weevil Hylobius abietis. We followed insect damage in two large progeny trials (52 open-pollinated families with 100–200 individuals per family and trial) naturally infested by the pine weevil. Pine weevils damaged between 17 and 48% of the planted seedlings depending on the trial and year, and mortality due to weevil damage was up to 11.4%. The results indicate significant genetic variation in resistance to the pine weevil, and importantly, the variation was highly consistent across trials irrespective of contrasting incidence levels. Individual heritability estimates for the different components of seedling resistance were consistently low, but family heritabilities were moderate (0.53 to 0.81). While forward selections and breeding for higher resistance seem not feasible, backwards selections of the best parent trees emerge as a putative alternative to reduce weevil damage. A positive genetic correlation between early growth potential and probability of being attacked by the weevil was also observed, but the relationship was weak and appeared only in one of the trials. Overall, results presented here open the door to a new attractive way for reducing damage caused by this harmful pest.

Non-consumptive effects are increasingly recognized as important in predator-herbivore interactions, with both fundamental and practical implications. Notably, feeding damage to conifer seedlings by pine weevils, Hylobius abietis (L.), which are serious pests in forest regeneration areas, can be decreased by interactions with ants (Formicidae). To probe the behavioural mechanisms underlying such effects, which have rarely been described in predator-herbivore systems, we investigated how the ants influence the weevils' behaviour around planted conifer seedlings in the field. Half of the seedlings were infested with aphids and attended by red wood ants Formica polyctena Forster, whereas the others were used as controls. We asked whether the ants' behaviour towards the weevils differs between these treatments, and whether the weevils' movement behaviour (locomotion, stationary activity and digging) is affected by the treatments and physical interactions with ants. The ants attacked pine weevils more frequently and for longer periods around ant-attended seedlings than around controls. Weevils' locomotion behaviour also increased markedly when attacked by ants. This may increase the likelihood of pine weevils moving away from, and causing less damage to, ant-attended seedlings. This study demonstrates how non-consumptive effects can be explained in terms of a herbivore's responses to a predator's aggressive behaviours.

Conifer defences can be induced by exogenous chemical elicitors, thereby reducing damage caused by bark-feeding insects. However, the insect behavioural mechanisms involved are poorly understood. Thus, effects of artificially induced plant defences on feeding behaviour of the pine weevil (Hylobius abietis), a serious forest pest, were examined to explore mechanisms involved in decision-making processes connected to feeding. To induce plant defences, we applied methyl jasmonate (MeJA), a naturally occurring plant hormone, to young Norway spruce (Picea abies) plants. The weevils’ feeding behaviour on plants with and without MeJA treatment was studied in both a no-choice and a choice laboratory experiment. MeJA treatment did not affect the initiation of feeding, but it affected the weevils’ subsequent feeding patterns. In the no-choice experiment, the only observed effect of its treatment was that it reduced the size of the initial feeding scars. In the choice experiment, it reduced both the numbers and sizes of the feeding scars and hence the total debarked area. Thus, the MeJA-induced resistance did not deter the pine weevils from attacking the spruce plants, but reduced the amounts they consumed at one place, which would reduce risks of girdling and plant death. This may be the behavioural mechanism behind the previously recorded increases in survival rates of MeJA-treated plants in the field.

Natural enemies attracted to plants may provide those plants with protection against herbivores but may also protect neighbouring plants, that is through associational resistance. Ant attendance may be an important mechanism for the occurrence of such effects because ants can reduce the damage caused by a wide variety of herbivorous insects. Ants have been shown, in a previous field experiment, to decrease the damage caused by the pine weevil, Hylobius abietis (L.) (Coleoptera: Curculionidae), a pest species that causes high seedling mortality in forest regeneration areas. In this study, we specifically tested whether seedlings planted close to ant-attended seedlings experience associational resistance. We did this under laboratory conditions using the ant species Lasius niger (L.) (Hymenoptera: Formicidae). The feeding damage by pine weevils was significantly reduced on seedlings attended by ants. The neighbouring seedlings, however, did not experience associational resistance. Nevertheless, some associational effects were observed as the number of weevils recorded on both ant-attended and neighbouring seedlings was significantly lower compared with ant-excluded seedlings.