Biography

I study plant-insect interactions, and particularly how conifers defend themselves against bark beetles and fungi. The work is highly interdisciplinary and I collaborate with chemists, molecular biologists and ecologists at NIBIO and abroad. The methods we use span from field experiments to chemical ecology and molecular biology. I have worked at NIBIO since 1992. I studied at the University of Oslo, where I graduated as Master of Science in ecology / entomology in 1992 and Dr. Scient. (PhD) in forest entomology in 1996. Since 2004, I have been an adjunct professor at the Norwegian University of Life Sciences (NMBU), where I teach forest entomology. Since 2018, I am also a member of the Plant Health Panel of the Norwegian Scientific Committee for Food and Environment (VKM).

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Abstract

Convergent evolution of semiochemical use in organisms from different Kingdoms is a rarely described phenomenon. Tree-killing bark beetles vector numerous symbiotic blue-stain fungi that help the beetles colonize healthy trees. Here we show for the first time that some of these fungi are able to biosynthesize bicyclic ketals that are pheromones and other semiochemicals of bark beetles. Volatile emissions of five common bark beetle symbionts were investigated by gas chromatography-mass spectrometry. When grown on fresh Norway spruce bark the fungi emitted three well-known bark beetle aggregation pheromones and semiochemicals (exo-brevicomin, endo-brevicomin and trans-conophthorin) and two structurally related semiochemical candidates (exo-1,3-dimethyl-2,9-dioxabicyclo[3.3.1]nonane and endo-1,3-dimethyl-2,9-dioxabicyclo[3.3.1]nonane) that elicited electroantennogram responses in the spruce bark beetle Ips typographus. When grown on malt agar with 13C D-Glucose, the fungus Grosmannia europhioides incorporated 13C into exo-brevicomin and trans-conophthorin. The enantiomeric compositions of the fungus-produced ketals closely matched those previously reported from bark beetles. The production of structurally complex bark beetle pheromones by symbiotic fungi indicates cross-kingdom convergent evolution of signal use in this system. This signaling is susceptible to disruption, providing potential new targets for pest control in conifer forests and plantations.

Abstract

1 The European spruce bark beetle Ips typographus is a damaging pest on spruce in Europe. Beetle interactions with tree species originating outside the natural range of the beetle are largely unknown and may be unpredictable because trees without a co-evolutionary history with the beetle may lack effective defences. 2 The terpenoid composition and breeding suitability for I. typographus of the historic host Norway spruce Picea abies were compared with two evolutionary naïve spruces of North American origin that are extensively planted in North-West Europe: Sitka spruce Picea sitchensis and Lutz spruce Picea glauca x lutzii. 3 The bark of all three species had a similar chemical composition and similar levels of total constitutive terpenoids, although Norway spruce had higher total induced terpenoid levels. 4 Beetles tunnelling in the three spruce species produced similar amounts of aggregation pheromone. Controlled breeding experiments showed that I. typographus could produce offspring in all three species, with a similar offspring length and weight across species. However, total offspring production was much lower in Sitka and Lutz spruce. 5 Overall, the results of the present study suggest that I. typographus will be able to colonize Sitka and Lutz spruce in European plantations and in native spruce forests in North America if introduced there.

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Abstract

The purpose of this study is to increase the basic understanding of outbreak dynamics in order to improve the management of bark beetle outbreaks. The spruce bark beetle Ips typographus is a major disturbance agent of European forests and is the continent’s most economically and environmentally damaging bark beetle. Outbreaks of the spruce bark beetle are often triggered by large windfall episodes, and we have utilized a unique opportunity to study a Slovakian outbreak where little salvage logging was performed in some areas after a 2.5 million m3 storm-felling in 2004. Our analyses focused on the first five years after the windfall, and we used a combination of empirical data and simulation models to understand the spatial patterns of beetle-killed forest patches developing during the outbreak. The univoltine beetle population used an increasing proportion of the windfelled trees during the two first seasons after the storm, but from the third season onwards our comparisons of inter-patch distance distributions indicated a transition from beetle production largely in windfall areas to a self-sustaining outbreak with infestation patches developing independently of the windthrows. The size of new infestation patches formed after this transition was modeled as a function of beetle pressure, estimated by the proportion of a circle area surrounding new patches that was covered by infestation patches the previous year. Our model results of patch size distribution did not correspond well with the empirical data if patch formation was modeled as a pure dispersal–diffusion process. However, beetle aggregation on individual trees appears to be important for patch development, since good correspondence with empirical data was found when beetle aggregation was incorporated in the modeled dispersal process. The strength of correspondence between the beetle aggregation model and the empirical data varied with the density of aggregation trees in the modeled landscape, and reached a maximum of 83% for a density of three aggregation trees per infestation patch. Our results suggest that efficient removal of windfelled trees up until the start of the second summer after a major windfall is important to avoid a transition into a patch-driven bark beetle outbreak that is very difficult to manage. Our results also indicate that the outcome of a patch-driven outbreak is difficult to predict, since the development of new infestation patches is not a simple function of beetle pressure but is also affected by beetle behavior and local forest conditions.

Abstract

Climate change is already reducing carbon sequestration in Central European forests dramatically through extensive droughts and bark beetle outbreaks. Further warming may threaten the enormous carbon reservoirs in the boreal forests in northern Europe unless disturbance risks can be reduced by adaptive forest management. The European spruce bark beetle (Ips typographus) is a major natural disturbance agent in spruce-dominated forests and can overwhelm the defences of healthy trees through pheromone-coordinated mass-attacks. We used an extensive dataset of bark beetle trap counts to quantify how climatic and management-related factors influence bark beetle population sizes in boreal forests. Trap data were collected during a period without outbreaks and can thus identify mechanisms that drive populations towards outbreak thresholds. The most significant predictors of bark beetle population size were the volume of mature spruce, the extent of newly exposed clearcut edges, temperature and soil moisture. For clearcut edge, temperature and soil moisture, a 3-year time lag produced the best model fit. We demonstrate how a model incorporating the most significant predictors, with a time lag, can be a useful management tool by allowing spatial prediction of future beetle population sizes. Synthesis and Applications: Some of the population drivers identified here, i,e., spruce volume and clearcut edges, can be targeted by adaptive management measures to reduce the risk of future bark beetle outbreaks. Implementing such measures may help preserve future carbon sequestration of European boreal forests.

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Abstract

Bark beetle (Ips typographus) outbreaks have the potential to damage large areas of spruce-dominated forests in Scandinavia. To define forest management strategies that will minimize the risk of bark beetle attacks, we need robust models that link forest structure and composition to the risk and potential damage of bark beetle attacks. Since data on bark beetle infestation rates and corresponding damages does not exist in Norway, we implement a previously published meta-model for estimating I. typographus damage probability and intensity. Using both current and projected climatic conditions we used the model to estimate damage inflicted by I. typographus in Norwegian spruce stands. The model produces feasible results for most of Norway’s climate and forest conditions, but a revised model tailored to Norway should be fitted to a dataset that includes older stands and lower temperatures. Based on current climate and forest conditions, the model predicts that approximately nine percent of productive forests within Norway’s main spruce-growing region will experience a loss ranging from 1.7 to 11 m3/ha of spruce over a span of five years. However, climate change is predicted to exacerbate the annual damage caused by I. typographus, potentially leading to a doubling of its detrimental effects.

Abstract

Purpose of Review Forestry in northern temperate and boreal regions relies heavily on conifers. Rapid climate change and associated increases in adverse growing conditions predispose conifers to pathogens and pests. The much longer generation time and presumably, therefore, lower adaptive capacity of conifers relative to their native or non-native biotic stressors may have devastating consequences. We provide an updated overview of conifer defences underlying pathogen and pest resistance and discuss how defence traits can be used in tree breeding and forest management to improve resistance. Recent Findings Breeding of more resilient and stress-resistant trees will benefit from new genomic tools, such as genotyping arrays with increased genomic coverage, which will aid in genomic and relationship-based selection strategies. However, to successfully increase the resilience of conifer forests, improved genetic materials from breeding programs must be combined with more flexible and site-specific adaptive forest management. Summary Successful breeding programs to improve conifer resistance to pathogens and pests provide hope as well as valuable lessons: with a coordinated and sustained effort, increased resistance can be achieved. However, mechanisms underlying resistance against one stressor, even if involving many genes, may not provide any protection against other sympatric stressors. To maintain the adaptive capacity of conifer forests, it is important to keep high genetic diversity in the tree breeding programs. Choosing forest management options that include diversification of tree-species and forest structure and are coupled with the use of genetically improved plants and assisted migration is a proactive measure to increase forest resistance and resilience to foreseen and unanticipated biotic stressors in a changing climate.

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Abstract

Drought-induced mortality is a major direct effect of climate change on tree health, but drought can also affect trees indirectly by altering their susceptibility to pathogens. Here, we report how a combination of mild or severe drought and pathogen infection affected the growth, pathogen resistance and gene expression in potted 5-year-old Norway spruce trees [Picea abies (L.) Karst.]. After 5 weeks of drought, trees were inoculated with the fungal pathogen Endoconidiophora polonica. Combined drought–pathogen stress over the next 8 weeks led to significant reductions in the growth of drought-treated trees relative to well-watered trees and more so in trees subjected to severe drought. Belowground, growth of the smallest fine roots was most affected. Aboveground, shoot diameter change was most sensitive to the combined stress, followed by shoot length growth and twig biomass. Both drought-related and some resistance-related genes were upregulated in bark samples collected after 5 weeks of drought (but before pathogen infection), and gene expression levels scaled with the intensity of drought stress. Trees subjected to severe drought were much more susceptible to pathogen infection than well-watered trees or trees subjected to mild drought. Overall, our results show that mild drought stress may increase the tree resistance to pathogen infection by upregulating resistance-related genes. Severe drought stress, on the other hand, decreased tree resistance. Because drought episodes are expected to become more frequent with climate change, combined effects of drought and pathogen stress should be studied in more detail to understand how these stressors interactively influence tree susceptibility to pests and pathogens.

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An epigenetic memory of the temperature sum experienced during embryogenesis is part of the climatic adaptation strategy of the long-lived gymnosperm Norway spruce. This memory has a lasting effect on the timing of bud phenology and frost tolerance in the resulting epitype trees. The epigenetic memory is well characterized phenotypically and at the transcriptome level, but to what extent DNA methylation changes are involved have not previously been determined. To address this, we analyzed somatic epitype embryos of Norway spruce clones produced at contrasting epitype-inducing conditions (18 and 28°C). We screened for differential DNA methylation in 2744 genes related mainly to the epigenetic machinery, circadian clock, and phenology. Of these genes, 68% displayed differential DNA methylation patterns between contrasting epitype embryos in at least one methylation context (CpG, CHG, CHH). Several genes related to the epigenetic machinery (e.g., DNA methyltransferases, ARGONAUTE) and the control of bud phenology (FTL genes) were differentially methylated. This indicates that the epitype-inducing temperature conditions induce an epigenetic memory involving specific DNA methylation changes in Norway spruce.

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Stress can have long-lasting impacts on plants. Here we report the long-term effects of the stress hormone jasmonic acid (JA) on the defence phenotype, transcriptome and DNA methylome of Arabidopsis. Three weeks after transient JA signalling, 5-week-old plants retained induced resistance (IR) against herbivory but showed increased susceptibility to pathogens. Transcriptome analysis revealed long-term priming and/or upregulation of JA-dependent defence genes but repression of ethylene- and salicylic acid-dependent genes. Long-term JA-IR was associated with shifts in glucosinolate composition and required MYC2/3/4 transcription factors, RNA-directed DNA methylation, the DNA demethylase ROS1 and the small RNA (sRNA)-binding protein AGO1. Although methylome analysis did not reveal consistent changes in DNA methylation near MYC2/3/4-controlled genes, JA-treated plants were specifically enriched with hypomethylated ATREP2 transposable elements (TEs). Epigenomic characterization of mutants and transgenic lines revealed that ATREP2 TEs are regulated by RdDM and ROS1 and produce 21 nt sRNAs that bind to nuclear AGO1. Since ATREP2 TEs are enriched with sequences from IR-related defence genes, our results suggest that AGO1-associated sRNAs from hypomethylated ATREP2 TEs trans-regulate long-lasting memory of JA-dependent immunity.

Abstract

A major challenge for plants in a rapidly changing climate is to adapt to rising temperatures. Some plants adapt to temperature conditions by generating an epigenetic memory that can be transmitted both meiotically and mitotically. Such epigenetic memories may increase phenotypic variation to global warming and provide time for adaptation to occur through classical genetic selection. The goal of this study was to understand how warmer temperature conditions experienced during sexual and asexual reproduction affect the transcriptomes of different strawberry (Fragaria vesca) ecotypes. We let four European F. vesca ecotypes reproduce at two contrasting temperatures (18 and 28°C), either asexually through stolon formation for several generations, or sexually by seeds (achenes). We then analyzed the transcriptome of unfolding leaves, with emphasis on differential expression of genes belonging to the epigenetic machinery. For asexually reproduced plants we found a general transcriptomic response to temperature conditions but for sexually reproduced plants we found less significant responses. We predicted several splicing isoforms for important genes (e.g. a SOC1, LHY, and SVP homolog), and found significantly more differentially presented splicing event variants following asexual vs. sexual reproduction. This difference could be due to the stochastic character of recombination during meiosis or to differential creation or erasure of epigenetic marks during embryogenesis and seed development. Strikingly, very few differentially expressed genes were shared between ecotypes, perhaps because ecotypes differ greatly both genetically and epigenetically. Genes related to the epigenetic machinery were predominantly upregulated at 28°C during asexual reproduction but downregulated after sexual reproduction, indicating that temperature-induced change affects the epigenetic machinery differently during the two types of reproduction.

Abstract

The large pine weevil (Hylobius abietis) is a major regeneration pest in commercial forestry. Pesticide application has historically been the preferred control method, but pesticides are now being phased out in several countries for environmental reasons. There is, thus, a need for alternative plant protection strategies. We applied methyl jasmonate (MeJA), salicylic acid (SA) or oxalic acid (OxA) on the stem of 2-year-old Norway spruce (Picea abies) plants to determine effects on inducible defenses and plant growth. Anatomical examination of stem cross-sections 9 weeks after application of 100 mM MeJA revealed massive formation of traumatic resin ducts and greatly reduced sapwood growth. Application of high concentrations of SA or OxA (500 and 200 mM, respectively) induced much weaker physiological responses than 100 mM MeJA. All three treatments reduced plant height growth significantly, but the reduction was larger for MeJA (~55%) than for SA and OxA (34-35%). Lower MeJA concentrations (5-50 mM) induced comparable traumatic resin duct formation as the high MeJA concentration but caused moderate (and non-significant) reductions in plant growth. Two-year-old spruce plants treated with 100 mM MeJA showed reduced mortality after exposure to pine weevils in the field, and this enhanced resistance-effect was statistically significant for three years after treatment.

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Temperature conditions experienced during embryogenesis and seed development may induce epigenetic changes that increase phenotypic variation in plants. Here we investigate if embryogenesis and seed development at two different temperatures (28 vs. 18°C) result in lasting phenotypic effects and DNA methylation changes in woodland strawberry (Fragaria vesca). Using five European ecotypes from Spain (ES12), Iceland (ICE2), Italy (IT4), and Norway (NOR2 and NOR29), we found statistically significant differences between plants from seeds produced at 18 or 28°C in three of four phenotypic features investigated under common garden conditions. This indicates the establishment of a temperature-induced epigenetic memory-like response during embryogenesis and seed development. The memory effect was significant in two ecotypes: in NOR2 flowering time, number of growth points and petiole length were affected, and in ES12 number of growth points was affected. This indicates that genetic differences between ecotypes in their epigenetic machinery, or other allelic differences, impact this type of plasticity. We observed statistically significant differences between ecotypes in DNA methylation marks in repetitive elements, pseudogenes, and genic elements. Leaf transcriptomes were also affected by embryonic temperature in an ecotype-specific manner. Although we observed significant and lasting phenotypic change in at least some ecotypes, there was considerable variation in DNA methylation between individual plants within each temperature treatment. This within-treatment variability in DNA methylation marks in F. vesca progeny may partly be a result of allelic redistribution from recombination during meiosis and subsequent epigenetic reprogramming during embryogenesis.

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The European spruce bark beetle Ips typographus and the North American spruce beetle Dendroctonus rufipennis cause high mortality of spruces on their native continents. Both species have been inadvertently transported beyond their native ranges. With similar climates and the presence of congeneric spruce hosts in Europe and North America, there is a risk that one or both bark beetle species become established into the non-native continent. There are many challenges that an introduced population of bark beetles would face, but an important prerequisite for establishment is the presence of suitable host trees. We tested the suitability of non-native versus native hosts by exposing cut bolts of Norway spruce (Picea abies), black spruce (Picea mariana) and white spruce (Picea glauca) to beetle attacks in the field in Norway and Canada. We quantified attack density, brood density and reproductive success of I. typographus and D. rufipennis in the three host species. We found that I. typographus attacked white and black spruce at comparable densities to its native host, Norway spruce, and with similar reproductive success in all three host species. In contrast, D. rufipennis strongly preferred to attack white spruce (a native host) but performed better in the novel Norway spruce host than it did in black spruce, a suboptimal native host. Our results suggest that I. typographus will find abundant and highly suitable hosts in North America, while D. rufipennis in Europe may experience reduced reproductive success in Norway spruce.

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Abstract

Key words: VKM, pest risk analysis, Norwegian Scientific Committee for Food and Environment, Norwegian Food Safety Authority, Sudden oak death, Phytophthora ramorum Introduction The Norwegian Food Safety Authority has asked the Norwegian Scientific Committee for Food and Environment for an updated pest risk assessment of Phytophthora ramorum in Norway. The previous risk assessment of P. ramorum for Norway is from 2009. Since then, the pathogen has been detected repeatedly in Norway, primarily in parks, garden centres, and nurseries in southwestern Norway. The knowledge base concerning P. ramorum has changed since the last pest risk assessment, with increased genetic knowledge about different populations, lineages, and mating types. The risks associated with P. ramorum have also changed, since the disease has become epidemic in new host plants, such as larch trees in England. This updated pest risk assessment will provide important input to the Norwegian Food Safety Authority’s efforts to develop the Norwegian plant health regulation. Methods VKM established a project group with expertise in plant health, forest pathology, horticultural plant pathology, plant disease modelling, and pest risk assessment. The group conducted systematic literature searches and scrutinized the relevant literature. In the absence of Norwegian studies, VKM relied on literature from other countries. The group did a quantitative risk assessment describing the level of confidence in the conclusions and identifying uncertainties and data gaps. The report underwent pre-submission commenting and external expert reviewing before final approval and publication. Results and conclusions Phytophthora ramorum is present in the PRA area but has a restricted distribution, mainly being detected in the southern and southwestern parts of Norway. The only P. ramorum lineage considered to be present in Norway is EU1 with mating type A1. The other lineage in Europe, EU2, has so far mainly been documented from the UK. The most widely distributed multilocus genotype of P. ramorum in Norway is EU1MLG1, which became dominant in Europe (including Norway) after 2008. In North America, the NA1, NA2, and EU1 lineages are known from both nurseries and forests. NA1 and NA2 are of the opposite mating type (A2) than European lineages. Recently, various other lineages of P. ramorum have been described from Asia. The main risks for future problems with P. ramorum in Norway are related to entry and establishment of non-European isolates (of all lineages), as well as emergence of new genotypes in European P. ramorum populations. There are several options for diagnosing P. ramorum to species and lineage (mainly EU1, EU2, NA1, and NA2). From a management perspective it is more important to distinguish these entities than mating type and isolate groups (genotypes). The latter are mainly relevant for research purposes or in cases of unexpected disease developments, such as new hosts, increased spread or more severe symptoms on known hosts. However, for more detailed regulation, monitoring, and management of P. ramorum it could also be useful to test for genotypes, i.e. to distinguish EU1MLG1 from other genotypes. Rhododendron remains the most important host plant for P. ramorum in Norway, both in terms of imported plants and detections (mainly in nurseries, garden centres, and public parks). Species in other ornamental plant genera, such as Viburnum, Pieris, and Kalmia, are also listed as major hosts in Europe, and P. ramorum has been detected at least once on species in all these genera in Norway. In the US, Rhododendron, Viburnum, Pieris, Syringa, and Camellia are considered to be the main ornamental hosts. .....................

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Plants must adapt with increasing speed to global warming to maintain their fitness. One rapid adaptation mechanism is epigenetic memory, which may provide organisms sufficient time to adapt to climate change. We studied how the perennial Fragaria vesca adapted to warmer temperatures (28°C vs. 18°C) over three asexual generations. Differences in flowering time, stolon number, and petiole length were induced by warmer temperature in one or more ecotypes after three asexual generations and persisted in a common garden environment. Induced methylome changes differed between the four ecotypes from Norway, Iceland, Italy, and Spain, but shared methylome responses were also identified. Most differentially methylated regions (DMRs) occurred in the CHG context, and most CHG and CHH DMRs were hypermethylated at the warmer temperature. In eight CHG DMR peaks, a highly similar methylation pattern could be observed between ecotypes. On average, 13% of the differentially methylated genes between ecotypes also showed a temperature-induced change in gene expression. We observed ecotype-specific methylation and expression patterns for genes related to gibberellin metabolism, flowering time, and epigenetic mechanisms. Furthermore, we observed a negative correlation with gene expression when repetitive elements were found near (±2 kb) or inside genes. In conclusion, lasting phenotypic changes indicative of an epigenetic memory were induced by warmer temperature and were accompanied by changes in DNA methylation patterns. Both shared methylation patterns and transcriptome differences between F. vesca accessions were observed, indicating that DNA methylation may be involved in both general and ecotype-specific phenotypic variation.

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Abstract

Tree-killing bark beetles in conifer forests vector symbiotic fungi that are thought to help the beetles kill trees. Fungal symbionts emit diverse volatile blends that include bark beetle semiochemicals involved in mating and host localization. In this study, all 12 tested fungal isolates emitted beetle semiochemicals when growing in medium amended with linoleic acid. These semiochemicals included the spiroacetals chalcogran, trans-conophthorin and exo-brevicomin, as well as 2-methyl-3-buten-1-ol, the main aggregation pheromone component of the spruce bark beetle Ips typographus. The emission of these compounds was affected by the type of fatty acid present (linoleic vs. oleic acid). Accumulating evidence shows that the fatty acid composition in conifer bark can facilitate colonization by bark beetles and symbiotic fungi, whereas the fatty acid composition of non-host trees can be detrimental for beetle larvae or fungi. We hypothesize that beetles probe the fatty acid composition of potential host trees to test their suitability for beetle development and release of semiochemicals by symbiotic fungi.

Abstract

Defense priming, the sensitization of inducible defenses, has been extensively studied in annual angiosperms. However, we are just beginning to explore defense priming in woody, long-lived plants. The natural compound methyl jasmonate (MeJA) has been used for over 20 years to study spruce inducible defenses. Recently, it was discovered that MeJA not only directly induces defense, but also primes defense responses in spruce. Metabolite and transcriptional analyses of mature trees treated with MeJA and subsequently wounded showed that while terpenes accumulate at the wound site in a primed manner, terpene biosynthesis genes are directly induced by MeJA. Pathogen resistance (PR) genes, on the other hand, are primed. Sequencing of miRNAs suggests that miRNAs have a regulatory role in MeJA-induced defenses in spruce. Additionally, a detailed transcriptional time course of 2- year-old spruce treated with MeJA indicated that the RNA-directed DNA methylation (RdDM) pathway is involved in the establishment and maintenance of primed defenses. When comparing mechanisms of defense priming in spruce to those in Arabidopsis, it seems that many mechanisms are conserved. However, some aspects, such as jasmonic acid-salicylic acid crosstalk, may be different. Identifying these differences and how they affect forest species is important for practical application of defense priming in forest management.

Abstract

Grey mold caused by the necrotrophic fungal pathogen Botrytis cinerea can affect leaves, flowers, and berries of strawberry, causing severe pre- and postharvest damage. The defense elicitor β-aminobutyric acid (BABA) is reported to induce resistance against B. cinerea and many other pathogens in several crop plants. Surprisingly, BABA soil drench of woodland strawberry (Fragaria vesca) plants two days before B. cinerea inoculation caused increased infection in leaf tissues, suggesting that BABA induce systemic susceptibility in F. vesca. To understand the molecular mechanisms involved in B. cinerea susceptibility in leaves of F. vesca plants soil drenched with BABA, we used RNA sequencing to characterize the transcriptional reprogramming 24 h post-inoculation. The number of differentially expressed genes (DEGs) in infected vs. uninfected leaf tissue in BABA-treated plants was 5205 (2237 upregulated and 2968 downregulated). Upregulated genes were involved in pathogen recognition, defense response signaling, and biosynthesis of secondary metabolites (terpenoid and phenylpropanoid pathways), while downregulated genes were involved in photosynthesis and response to auxin. In control plants not treated with BABA, we found a total of 5300 DEGs (2461 upregulated and 2839 downregulated) after infection. Most of these corresponded to those in infected leaves of BABA-treated plants but a small subset of DEGs, including genes involved in ‘response to biologic stimulus‘, ‘photosynthesis‘ and ‘chlorophyll biosynthesis and metabolism’, differed significantly between treatments and could play a role in the induced susceptibility of BABA-treated plants.

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Norway spruce (Picea abies) is an economically and ecologically important tree species that grows across northern and central Europe. Treating Norway spruce with jasmonate has long-lasting beneficial effects on tree resistance to damaging pests, such as the European spruce bark beetle Ips typographus and its fungal associates. The (epi)genetic mechanisms involved in such long-lasting jasmonate induced resistance (IR) have gained much recent interest but remain largely unknown. In this study, we treated 2-year-old spruce seedlings with methyl jasmonate (MeJA) and challenged them with the I. typographus vectored necrotrophic fungus Grosmannia penicillata. MeJA treatment reduced the extent of necrotic lesions in the bark 8 weeks after infection and thus elicited long-term IR against the fungus. The transcriptional response of spruce bark to MeJA treatment was analysed over a 4-week time course using mRNA-seq. This analysis provided evidence that MeJA treatment induced a transient upregulation of jasmonic acid, salicylic acid and ethylene biosynthesis genes and downstream signalling genes. Our data also suggests that defence-related genes are induced while genes related to growth are repressed by methyl jasmonate treatment. These results provide new clues about the potential underpinning mechanisms and costs associated with long-term MeJA-IR in Norway spruce.

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Invasive species are leading causes of biodiversity loss and economic damage. Prevention and management of invasions requires risk assessments based on ecological knowledge for species of potential concern. Interactions between introduced species and heterospecifics in the recipient community may affect the likelihood of establishment through biotic resistance and facilitation and are therefore important predictors of invasion risk. Experimentally exposing one species to another to observe their interactions is not always safe or practical, and containment facilities offer artificial environments which may limit the number of species and the types of interactions that may be tested. To predict biotic resistance and facilitation in a more natural setting, we deployed traps with pheromone lures in the field to mimic the presence of two potentially invasive spruce bark beetles, the European Ips typographus (tested in eastern Canada), and the North American Dendroctonus rufipennis (tested in Norway). We identified and counted possible predators, competitors, and facilitators that were captured in the traps. In eastern Canada, possible predators and competitors responded strongly to I. typographus lures, suggesting the potential for considerable biotic resistance. In Norway, D. rufipennis lures prompted little response by predators or competitors, suggesting that D. rufipennis may experience reduced biotic resistance in Europe. Dendroctonus rufipennis was also attracted to I. typographus pheromone, which may encourage facilitation between these species through cooperative mass attack on trees. Our findings will inform invasive-species risk assessments for I. typographus and D. rufipennis and highlight useful methods for predicting interactions between species that rely heavily on semiochemical communication.

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Abstract

Conifer-feeding bark beetles are important herbivores and decomposers in forest ecosystems. These species complete their life cycle in nutritionally poor substrates and some can kill enormous numbers of trees during population outbreaks. The Eurasian spruce bark beetle (Ips typographus) can destroy >100 million m3 of spruce in a single year. We report a 236.8 Mb I. typographus genome assembly using PacBio long-read sequencing. The final phased assembly has a contig N50 of 6.65 Mb in 272 contigs and is predicted to contain 23,923 protein-coding genes. We reveal expanded gene families associated with plant cell wall degradation, including pectinases, aspartyl proteases, and glycosyl hydrolases. This genome sequence from the genus Ips provides timely resources to address questions about the evolutionary biology of the true weevils (Curculionidae), one of the most species-rich animal families. In forests of today, increasingly stressed by global warming, this draft genome may assist in developing pest control strategies to mitigate outbreaks.

Abstract

Key words: VKM, risk assessment, Norwegian Scientific Committee for Food and Environment, Norwegian Food Safety Authority, Norwegian Environment Agency, Biowaste, Compost, Plant health, organic waste, Phytosanitary safety, Biogas, Alien organisms Introduction The Norwegian Food Safety Authority (NFSA) and the Norwegian Environment Agency (NEA) have jointly asked the Norwegian Scientific Committee for food and environment for an assessment into treatment methods and validation methods for compost and digestate based on organic waste in relation to plant health and the spread of harmful alien organisms in Norway.  The Norwegian Food Safety Authority will use the report in its supervisory work over companies that produce compost and digestate. The assessment will also provide important input for the regulatory development of several current regulations including regulations on indicator organisms that are used to validate new methods and ensure adequate security with regards to the survival of plant pests. The Norwegian Environment Agency wants to establish whether the methods used in the composting of garden waste and other types of plant waste today are able to ensure that the finished product does not become a source for the spread of harmful alien organisms. This will form the basis for the Norwegian Environment Agency’s guidelines relating to the precautionary provisions in the regulation on alien organisms. This request is limited to an assessment of plant pests and harmful alien organisms (hereinafter alien organisms). The survival of infectious diseases harmful to people and animals is considered in separate assessments. Methods We have conducted initiating workshops for identifying relevant fundamental processes and parameters, of relevant organisms and of relevant search terms for the literature surveys, as well as for discussion and validation. Visits to composting facilities and contact with stakeholders in Norway were also conducted. This information was further implemented in an extensive literature search. This assessment include/encompass organic waste and other materials that are currently treated in biogas and composting facilities, including garden and park waste (incl. soil), plant waste from garden centres, etc., food waste and waste from the food and animal feed industry (including grain/seed husks and waste from enterprises which package and process potatoes and vegetables), manure, bulking agents used in composting facilities, and husks from contracted grain/seed cleaners for sowing. We have used a quantitative risk assessment. The level of confidence in the risk assessment is described, and uncertainties and data gaps identified. Furthermore, we have used re-submission commenting and external expert reviewing before final approval and publication. ...........

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Purpose of Review Outbreaks of tree-killing bark beetles have reached unprecedented levels in conifer forests in the northern hemisphere and are expected to further intensify due to climate change. In parts of Europe, bark beetle outbreaks and efforts to manage them have even triggered social unrests and political instability. These events have increasingly challenged traditional responses to outbreaks, and highlight the need for a more comprehensive management framework. Recent Findings Several synthesis papers on different aspects of bark beetle ecology and management exist. However, our understanding of outbreak drivers and impacts, principles of ecosystem management, governance, and the role of climate change in the dynamics of ecological and social systems has rapidly advanced in recent years. These advances are suggesting a reconsideration of previous management strategies. Summary We synthesize the state of knowledge on drivers and impacts of bark beetle outbreaks in Europe and propose a comprehensive context-dependent framework for their management. We illustrate our ideas for two contrasting societal objectives that represent the end-members of a continuum of forest management goals: wood and biomass production and the conservation of biodiversity and natural processes. For production forests, we propose a management approach addressing economic, social, ecological, infrastructural, and legislative aspects of bark beetle disturbances. In conservation forests, where non-intervention is the default option, we elaborate under which circumstances an active intervention is necessary, and whether such an intervention is in conflict with the objective to conserve biodiversity. Our approach revises the current management response to bark beetles in Europe and promotes an interdisciplinary social-ecological approach to dealing with disturbances.

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Nursery-grown Norway spruce Picea abies seedlings are often heavily attacked by the pine weevil Hylobius abietis on clear-cuts the first years after planting. Because the seedlings are not resource-limited during the growing phase in the nursery they are expected to invest less in defence than naturally regenerated seedlings already present on the clear-cuts. The latter have had to cope with various environmental stressors that could make them invest more in defence. We tested if naturally regenerated plants have stronger chemical defences than nursery-grown plants. Nursery-grown plants were planted in-between naturally regenerated plants on fresh clear cuts, and phenolic and terpene compounds in the stem bark were measured after one growing season. To test both constitutive and inducible defences, plants were either wounded, painted with methyl jasmonate (MeJA) to induce defences, or given a combination of both treatments. Growth and pine weevil attacks of the plants were registered. Nursery-grown plants had higher total concentrations of phenolic compounds and lower concentrations of terpenes than naturally regenerated plants. These opposite responses were reflected in very different compound profiles in the two plant types. We suggest the differences between plant types to be results of differences in plant age, stress level, genetic origin or possibly a combination of these factors. Most compounds showed no response to wounding, MeJA-treatment or wounding and MeJA-treatment combined, but the terpenes 3-carene, eucalyptol, limonene and para-cymene had higher concentrations in MeJA-treated nursery-grown plants than in control plants. These compounds are known to be effective in conifer resistance against weevils and bark beetles. Overall, 27% of our 400 study plants had signs of pine weevil damage after 3 ½ months in the field. However, treatment or plant type had no significant effect on whether plants were attacked or not and this might have been a result of the relatively low overall level of attacks in this study. Further studies are needed to disentangle the importance of plant age, stress level, genetic origin and resource availability for chemical defence mechanisms of young Norway spruce plants, as strengthening the natural resistance of nursery plants may be increasingly important in a future with less pesticide use.

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Abstract

The spruce bark beetle Ips typographus is the most damaging pest in European spruce forests and has caused great ecological and economic disturbances in recent years. Although native to Eurasia, I. typographus has been intercepted more than 200 times in North America and could establish there as an exotic pest if it can find suitable host trees. Using in vitro bioassays, we compared the preference of I. typographus for its coevolved historical host Norway spruce (Picea abies) and two non-coevolved (naïve) North American hosts: black spruce (Picea mariana) and white spruce (Picea glauca). Additionally, we tested how I. typographus responded to its own fungal associates (conspecific fungi) and to fungi vectored by the North American spruce beetle Dendroctonus rufipennis (allospecific fungi). All tested fungi were grown on both historical and naïve host bark media. In a four-choice Petri dish bioassay, I. typographus readily tunneled into bark medium from each of the three spruce species and showed no preference for the historical host over the naïve hosts. Additionally, the beetles showed a clear preference for bark media colonized by fungi and made longer tunnels in fungus-colonized media compared to fungus-free media. The preference for fungus-colonized media did not depend on whether the medium was colonized by conspecific or allospecific fungi. Furthermore, olfactometer bioassays demonstrated that beetles were strongly attracted toward volatiles emitted by both con- and allospecific fungi. Collectively, these results suggest that I. typographus could thrive in evolutionary naïve spruce hosts if it becomes established in North America. Also, I. typographus could probably form and maintain new associations with local allospecific fungi that might increase beetle fitness in naïve host trees.

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Abstract

Key words: VKM, risk assessment, Norwegian Scientific Committee for Food and Environment, Norwegian Food Safety Authority, biological control, Nematodes, Phasmarhabditis californica, Moraxella osloensis. Parasitic nematodes and associated bacteria are increasingly being used for biocontrol of molluscs. Functionally, it is the bacteria that kill and thus control the targeted pests, but the function of the bacteria is dependent on the nematodes, which should be regarded as vectors of biocontrol. Although the nematodes and the bacteria have a symbiotic relationship within such biocontrol formulations, it should be noted that they are not dependent on each other in the wild, but can establish separate populations which can be free-living or hosted by other organisms. The biocontrol product Nemaslug 2.0 contains the nematode Phasmarhabditis californica (strain P19D) and the bacterial symbiont Moraxella osloensis (unknown strain). The nematode was first described in 2016 and has never been reported in Norway. The lack of reports suggests that it is absent from Norway, but this conclusion comes with a high degree of uncertainty since there have been limited search efforts. The climatic thresholds of the nematode are not known, but its current distribution, spanning widely varying climates, suggests that it could survive and establish in Norway. Natural spread from currently known areas of establishment to natural habitats in Norway is ruled out due to the nematode’s limited dispersal capacity. However, human-assisted spread (e.g. via the use of biocontrol products) and establishment would be likely if Nemaslug 2.0 is allowed for use in open fields in Norway. Use of Nemaslug 2.0 in greenhouses and other enclosed areas is not likely to facilitate spread to natural habitats in Norway provided that residues are properly handled. However, deposition of product residues from greenhouses to outdoor areas may result in local establishment of the nematode in the vicinity of the deposition. Phasmarhabditis californica has a broad host range and may parasitize both rare/endangered and common mollusc species. However, there is no scientific evidence suggesting that the nematode can affect natural populations of molluscs in wild habitats, or otherwise have negative effects on biodiversity. The nematodes’ association with the bacteria Moraxella osloensis is most likely lost, or at least weakened, in natural habitats, suggesting that the nematode becomes less capable of killing its hosts in the wild. Phasmarhabditis californica is not capable of harming or infecting humans. The bacterial species Moraxella osloensis is already present in Norway in a few locations and at a low abundance, and it may be native to Norway. Little is known regarding its distribution in natural environments, but the literature shows that it can infect humans and other mammals. In humans with immunodeficiency or other comorbidities, M. osloensis can cause meningitis, vaginitis, sinusitis, bacteremia, endocarditis, and septic arthritis. The risk of infection in people handling Nemaslug 2.0 can probably be substantially reduced by protective clothing and appropriate handling. We are not aware of any reported health issues arising from use of the previous version of Nemaslug, which also contains M. osloensis. Different strains of M. osloensis are known to vary in their sensitivity to antibiotics, and likely in other traits too. Thus, the lack of information provided about the strain identity and specific characteristics of the strain used in Nemaslug 2.0 generates a high degree of uncertainty regarding its pathogenicity, climate tolerance, sensitivity to antibiotics etc.

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Abstract

In 2018, up to 4 million m3 Norway spruce was killed by the spruce bark beetle Ips typographus in Sweden. The event was unique for Sweden, in terms of both affected volume and the fact that it was triggered by severe drought stress, not by ample availability of relatively defenseless storm-felled trees. The outbreak continued in 2019 and 2020, each year with twice as many trees killed as in 2018. The aim of this study was to quantify seasonal variation and potential lag-effects in tree defense capacity the year after a severe drought stress. Inoculation with a bark beetle-associated bluestain fungus, repeated four times with one-month-intervals between May and August 2019, were carried out at three field sites with spruce provenances of Swedish and East European origin representing early and late bud burst, respectively. All sites had experienced moderate to severe drought stress in 2018, and site-specific defense capacity correlated positively with the cumulative precipitation two months before inoculation. Sites with two-month precipitation levels <100 mm had larger necrotic lesions in the phloem following inoculation, an indication of lower tree defense capacity. Lesion size did not differ between provenances, and all trees were able to confine fungal infection successfully. There were some seasonal differences in necrotic lesion size, with the sites Skärsnäs and Norberg having significantly larger lesions in June than in May, and site Lugnet having large lesions also in May. Lesions were generally smaller in July and August than in June. The cross-sectional area and number of traumatic resin ducts was measured in sapwood samples from one site, Lugnet, to quantify an additional aspect of tree defenses. The area of resin ducts produced in May and June were larger than that in July and August. This is in line with a positive correlation between lesion area and resin duct area, indicating that a stronger fungal infection following inoculation in spring triggered a stronger induced defense response. The East European provenances had more resin ducts than Swedish provenances, but the area of resin ducts did not differ significantly between provenances.

Abstract

Methyl jasmonate (MeJA) treatment elicits induced resistance (IR) against pests and diseases in Norway spruce (Picea abies). We recently demonstrated using mRNA-seq that this MeJA-IR is associated with both a prolonged upregulation of inducible defenses and defense priming. Gene expression can be regulated at both a transcrip-tional and post-transcriptional level by small RNAs, including microRNAs (miRNAs). Here we explore the effects of MeJA treatment and subsequent challenge by wounding on the Norway spruce miRNA transcriptome. We found clusters of prolonged down- or upregulated miRNAs as well as miRNAs whose expression was primed after MeJA treatment and subsequent wounding challenge. Differentially expressed miRNAs included miR160, miR167, miR172, miR319, and the miR482/2118 superfamily. The most prominent mRNA targets predicted to be differentially expressed by miRNA activity belonged to the nucleotide-binding site leucine-rich repeat (NBS- LRR) family. Among other predicted miRNA targets were genes regulating jasmonic acid biosynthesis. Our re-sults indicate that miRNAs have an important role in the regulation of MeJA-IR in Norway spruce.

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Abstract

In response to various stimuli, plants acquire resistance against pests and/or pathogens. Such acquired or induced resistance allows plants to rapidly adapt to their environment. Spraying the bark of mature Norway spruce (Picea abies) trees with the phytohormone methyl jasmonate (MeJA) enhances resistance to tree‐killing bark beetles and their associated phytopathogenic fungi. Analysis of spruce chemical defenses and beetle colonization success suggests that MeJA treatment both directly induces immune responses and primes inducible defenses for a faster and stronger response to subsequent beetle attack. We used metabolite and transcriptome profiling to explore the mechanisms underlying MeJA‐induced resistance in Norway spruce. We demonstrated that MeJA treatment caused substantial changes in the bark transcriptional response to a triggering stress (mechanical wounding). Profiling of mRNA expression showed a suite of spruce inducible defenses are primed following MeJA treatment. Although monoterpenes and diterpene resin acids increased more rapidly after wounding in MeJA‐treated than control bark, expression of their biosynthesis genes did not. We suggest that priming of inducible defenses is part of a complex mixture of defense responses that underpins the increased resistance against bark beetle colonization observed in Norway spruce. This study provides the most detailed insights yet into the mechanisms underlying induced resistance in a long‐lived gymnosperm.

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Abstract

Plants can form an immunological memory known as defense priming, whereby exposure to a priming stimulus enables quicker or stronger response to subsequent attack by pests and pathogens. Such priming of inducible defenses provides increased protection and reduces allocation costs of defense. Defense priming has been widely studied for short‐lived model plants such as Arabidopsis, but little is known about this phenomenon in long‐lived plants like spruce. We compared the effects of pretreatment with sublethal fungal inoculations or application of the phytohormone methyl jasmonate (MeJA) on the resistance of 48‐year‐old Norway spruce (Picea abies) trees to mass attack by a tree‐killing bark beetle beginning 35 days later. Bark beetles heavily infested and killed untreated trees but largely avoided fungus‐inoculated trees and MeJA‐treated trees. Quantification of defensive terpenes at the time of bark beetle attack showed fungal inoculation induced 91‐fold higher terpene concentrations compared with untreated trees, whereas application of MeJA did not significantly increase terpenes. These results indicate that resistance in fungus‐inoculated trees is a result of direct induction of defenses, whereas resistance in MeJA‐treated trees is due to defense priming. This work extends our knowledge of defense priming from model plants to an ecologically important tree species.

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Abstract

Key words: VKM, risk assessment, Norwegian Scientific Committee for Food and Environment, Norwegian Environment Agency, mycorrhiza. Mycorrhiza is a beneficial association between plant roots and fungi. This mutualistic symbiosis is essential for plant growth in most natural terrestrial ecosystems and in agriculture. Commercial mycorrhizal products containing fungi and bacteria may promote plant growth, especially on sites without a natural microbial community. Due to the risk of unintended negative effects, introduction of new species or genetically different isolates of native species should always be considered carefully. This report assesses the risk of establishment and spread of six fungal species and six bacterial species included in different commercial mycorrhizal products, as well as the species’ potential impact on Norwegian biodiversity. Most of the evaluated fungi and bacteria are probably present in Norway, even though presence at present data only exist for two of the six fungal species. Establishment of the assessed fungi on the plants and sites where they are applied is considered moderately likely, with medium uncertainty, while establishment of the bacterial species is considered to range from very unlikely to very likely depending on the bacterial group, with low uncertainty. The probability of spread to the wider environment ranges from unlikely (four fungal species), to moderately likely (two fungal species), to very likely (five of the six bacterial species). However, for all species it is considered unlikely that establishment and spread would have negative effects on other native species, habitats and ecosystems in Norway.

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Abstract

ANDERcontrol with the predatory mite Amblyseius andersoni as the active organism is sought to be used as a biological control agent in Norway. ANDERcontrol is intended for use against different mites (such as the two-spotted, fruit-tree, and red spider mite, russet mite,cyclamen mite) and in horticultural crops such as fruits, berries, vegetables, and ornamental. VKM’s conclusions are as follows Prevalence, especially if the organism is found naturally in Norway: Amblyseius andersoni has not been observed in Norway. It has been observed, in low numbers, in southern Sweden and has the capability to enter diapause under unfavourable conditions which suggests the potential for establishing under Norwegian conditions. It is however, the view of VKM that it likely lacks the ability to survive and establish in areas with cold winters and chilly summers, as found in most parts of Norway under current climatic conditions. The potential of the organism for establishment and spread under Norwegian conditions specified for use in greenhouses and open field: The thermal preference of A. andersoni restricts its establishment, and the species has not been observed in Norway. The species is capable of entering diapause, but the lack of records, despite targeted surveys, makes it the opinion of VKM that it is unlikely that A. andersoni will be able to establish in outdoor areas in Norway. However, the lack of information on temperature tolerance of the species constitute an uncertainty factor. The risk of spread from greenhouses is low because no wind or vector are likely to carry the mites from the greenhouse to suitable outdoor habitats, and mite populations in greenhouses do not enter the more cold-tolerant diapause. All conclusions are uncertain due to lack of relevant information regarding the species’ climate tolerance. Any ambiguities regarding the taxonomy, which hampers risk assessment: There are no taxonomic challenges related to the assessment of A. andersoni. Assessment of the product and the organism with regard to possible health risk: VKM is unaware of reports where harm to humans by A. andersoni itself, or associated pathogenic organisms have been observed. Mites may however produce allergic reactions in sensitive individuals handling plant material with high numbers of individuals. There is reason to believe that this holds true also for A. andersoni. Key words: VKM, risk assessment, Norwegian Scientific Committee for Food and Environment, Norwegian Food Safety Authority, biological control, predatory mite

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Abstract

The product Limonica, with the predatory mite Amblydromalus limonicus as the active organism, is sought to be used as a biological control agent in Norway. Limonica is intended for use against western flower thrips (Frankliniella occidentallis), other thrips (e.g. Thrips tabaci), spider mites and whiteflies (e.g. Trialeurodes, Aleyrodes and Bemisia spp.) in protected horticultural crops such as cucumber, sweet pepper, strawberry and ornamentals. The product is not recommended for greenhouse-grown tomatoes. VKM’s conclusions are as follows Distribution, especially if the organism is found naturally in Norway Amblydromalus limonicus has a very wide natural distribution, being reported from New Zealand, Australia South America, Central America, and North America as well as Hawaii. It has also recently established populations in crop productions and non-crop vegetation in Catalonia, North Eastern Spain. Amblydromalus limonicus have not been observed in Norway. The species seems not to have the capability to enter diapause under unfavourable conditions and it is the view of VKM that it likely lacks the ability to survive and establish in areas with cold winters and chilly summers, as found in most parts of Norway under current climatic conditions. The potential of the organism for establishment and spread under Norwegian conditions specified for use in greenhouses and open field The thermal preference of A. limonicus restricts its establishment, and the species has not been observed outdoors in Norway. As the species is incapable of entering diapause it is the opinion of VKM that it is unlikely that A. limonicus will be able to establish in outdoor areas in Norway. However, the lack of detailed information on temperature tolerance of the species constitutes an uncertainty factor. The risk of spread from greenhouses is low because no wind or vector are likely to carry the mites from the greenhouse to suitable outdoor habitats. However, mites that have escaped from a greenhouses to may spread in the nature. All conclusions are uncertain due to lack of relevant information regarding the species’ climate tolerance. Its origin and current distribution suggest that it cannot survive cold winters. Any ambiguities regarding taxonomy that hamper risk assessment There are no taxonomic challenges related to the assessment of A. limonicus. Assessment of the product and the organism with regard to possible health risks VKM Report 2020: 13 8 VKM is unaware of reports where harm to humans has been observed, whether by A. limonicus itself. Mites may, however, produce allergic reactions in sensitive individuals handling plant material with high numbers of individuals. There is reason to believe that this holds true also for A. limonicus. Key words: VKM, risk assessment, Norwegian Scientific Committee for Food and Environment, Norwegian Food Safety Authority, biological control, predatory mite

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Abstract

Atheta-System with the rove beetle Atheta coriaria (Kraatz 1856) as the active organism is sought to be used as a biocontrol agent for augmentation biological control in Norway. Atheta-System is intended for use against soil dwelling stages of fungus gnats (e.g. Bradysia paupera), shore flies (Scatella stagnalis), and thrips (e.g. Frankliniella occidentallis) in greenhouses, plastic tunnels, and other closed or controlled climate cultivations of horticultural crops, incl. soft-fruit crops, vegetables, ornamentals, and kitchen herbs. VKM’s conclusions are as follows Distribution, especially if the organism is found naturally in Norway Atheta coriaria is established (naturalized) in Norway since 1919 and has been reported numerous times from Agder in the South to Trøndelag in mid-Norway. The potential of the organism for establishment and spread under Norwegian conditions specified for use in greenhouses and open field The thermal thresholds of A. coriaria are not well-studied, but its current distribution in Southern and mid-Scandinavia shows that it tolerates relatively low winter temperatures, and that the Norwegian summer climate allows for successful reproduction. A. coriaria overwinters in the soil, which provides a relatively sheltered environment. Adults disperse rapidly by flying. All life stages can be vectored by humans – mainly by movement of soil and compost material. Thus, further spread northwards in Norway is predicted irrespective of additional introductions. It is unknown if it can enter diapause under greenhouse conditions. Any ambiguities regarding the taxonomy which hamper risk assessment There are no major taxonomic challenges related to the assessment of A. coriaria. Assessment of the product and the organism with regard to possible health risk VKM is unaware of reports of harm inflicted to humans by A. coriaria itself. Atheta-System comes with the cosmopolitan cheese mite (Tyrophagus putrescentiae), serving as food for A. coriaria. As with most mites, T. putrescentiae may induce allergic reactions in sensitive persons handling the product. Key words: VKM, risk assessment, Norwegian Scientific Committee for Food and Environment, Norwegian Food Safety Authority, biological control, rove beetle

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Abstract

Tree-killing bark beetles are the most economically important insects in conifer forests worldwide. However, despite >200 years of research, the drivers of population eruptions and crashes are still not fully understood and the existing knowledge is thus insufficient to face the challenges posed by the Anthropocene. We critically analyze potential biotic and abiotic drivers of population dynamics of an exemplary species, the European spruce bark beetle (ESBB) (Ips typographus) and present a multivariate approach that integrates the many drivers governing this bark beetle system. We call for hypothesis-driven, large-scale collaborative research efforts to improve our understanding of the population dynamics of this and other bark beetle pests. Our approach can serve as a blueprint for tackling other eruptive forest insects.

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Abstract

Convergent evolution of semiochemical use in organisms from different Kingdoms is a rarely described phenomenon. Tree-killing bark beetles vector numerous symbiotic blue-stain fungi that help the beetles colonize healthy trees. Here we show for the first time that some of these fungi are able to biosynthesize bicyclic ketals that are pheromones and other semiochemicals of bark beetles. Volatile emissions of five common bark beetle symbionts were investigated by gas chromatography-mass spectrometry. When grown on fresh Norway spruce bark the fungi emitted three well-known bark beetle aggregation pheromones and semiochemicals (exo-brevicomin, endo-brevicomin and trans-conophthorin) and two structurally related semiochemical candidates (exo-1,3-dimethyl-2,9-dioxabicyclo[3.3.1]nonane and endo-1,3-dimethyl-2,9-dioxabicyclo[3.3.1]nonane) that elicited electroantennogram responses in the spruce bark beetle Ips typographus. When grown on malt agar with 13C D-Glucose, the fungus Grosmannia europhioides incorporated 13C into exo-brevicomin and trans-conophthorin. The enantiomeric compositions of the fungus-produced ketals closely matched those previously reported from bark beetles. The production of structurally complex bark beetle pheromones by symbiotic fungi indicates cross-kingdom convergent evolution of signal use in this system. This signaling is susceptible to disruption, providing potential new targets for pest control in conifer forests and plantations.

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Abstract

Several studies have looked at how individual environmental factors influence needle morphology in conifer trees, but interacting effects between drought and canopy position have received little attention. In this study, we characterized morphological responses to experimentally induced drought stress in sun exposed and shaded current-year Norway spruce needles. In the drought plot trees were suffering mild drought stress, with an average soil water potential at 50 cm depth of -0.4 MPa. In general, morphological needle traits had greater values in sun needles in the upper canopy than in shaded needles in the lower canopy. Needle morphology 15 months after the onset of drought was determined by canopy position, as only sun needle morphology was affected by drought. Thus, canopy position was a stronger morphogenic factor determining needle structure than was water availability. The largest influence of mild drought was observed for needle length, projected needle area and total needle area, which all were reduced by ~27% relative to control trees. Needle thickness and needle width showed contrasting sensitivity to drought, as drought only affected needle thickness (10% reduction). Needle dry mass, leaf mass per area and needle density were not affected 15 months after the onset of mild drought. Our results highlight the importance of considering canopy position as well as water availability when comparing needle structure or function between conifer species. More knowledge about how different canopy parts of Norway spruce adapt to drought is important to understand forest productivity under changing environmental conditions.

Abstract

Plants are exposed to various pathogens in their environment and have developed immune systems with multiple defense layers to prevent infections. However, often pathogens overcome these resistance barriers, infect plants and cause disease. Pathogens that cause diseases on economically important crop plants incur huge losses to the agriculture industry. For example, the 2016 outbreak of strawberry grey mold (Botrytis cinerea) in Norway caused up to 95% crop losses. Such outbreaks underline the importance of developing novel and sustainable tools to combat plant diseases, for example by increasing the plants’ natural disease resistance. Priming plant defenses using chemical elicitors may enhance resistance against multiple pathogens. Such an approach may reduce the use of chemical fungicides and pesticides that often select for resistant strains of pests and pathogens. My presentation will focus on the effectiveness of different chemical agents to prime woodland strawberry (Fragaria vesca) defenses against the necrotroph B. cinerea. We have identified several genes that seem to play a role in disease resistance in strawberry and associated epigenetic memory mechanisms. Our results point out new management avenues for more sustainable crop protection schemes.

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Abstract

As primary producers, plants are under constant pressure to defend themselves against potentially deadly pathogens and herbivores. In this review, we describe short- and long-term strategies that enable plants to cope with these stresses. Apart from internal immunological strategies that involve physiological and (epi)genetic modifications at the cellular level, plants also employ external strategies that rely on recruitment of beneficial organisms. We discuss these strategies along a gradient of increasing timescales, ranging from rapid immune responses that are initiated within seconds to (epi)genetic adaptations that occur over multiple plant generations. We cover the latest insights into the mechanistic and evolutionary underpinnings of these strategies and present explanatory models. Finally, we discuss how knowledge from short-lived model species can be translated to economically and ecologically important perennials to exploit adaptive plant strategies and mitigate future impacts of pests and diseases in an increasingly interconnected and changing world.

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Abstract

Yunnan pine is the most important tree species in SW China in both economical and ecological terms, but it is often killed by pine shoot beetles (Tomicus spp.). Tomicus beetles are secondary pests in temperate regions and the aggressiveness of the beetles in SW China is considered to be due to the warm subtropical climates as well as the beetles’ virulent fungal associates. Here, we assessed the virulence of three blue-stain fungi (Leptographium wushanense, L. sinense and Ophiostoma canum) associated with pine shoot beetles to Yunnan pine (Pinus yunnanensis) in SW China. Following fungal inoculation, we measured necrotic lesion lengths, antioxidant enzyme activities and monoterpene concentrations in the stem phloem of Yunnan pine. Leptographium wushanense induced twice as long lesions as L. sinense and O. canum, and all three fungi induced significantly longer lesions than sterile agar control inoculations. The activity of three tested antioxidant enzymes (peroxidase, polyphenol oxidase, and superoxide dismutase) increased after both fungal inoculation and control inoculation. However, L. wushanense and L. sinense generally caused a greater increase in enzyme activities than O. canum and the control treatment. Fungal inoculation induced stronger increases in six major monoterpenes than the control treatment, but the difference was significant only for some fungus-monoterpene combinations. Overall, our results show that L. wushanense and L. sinense elicit stronger defense responses and thus are more virulent to Yunnan pine than O. canum. The two Leptographium species may thus contribute to the aggressiveness of their beetle vectors and could damage Yunnan pine across SW China if they spread from the restricted geographical area they have been found in so far.

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Abstract

Climate change is one of the greatest challenges for the biosphere. As sessile organisms, plants must adapt quickly to keep pace with the rapidly changing climatic conditions. Epigenetic memory is one mechanism which would provide sufficient plasticity under rapid climate change and enable long-lived organisms to survive long enough to adapt by classical genetic selection. In Norway spruce, the timing of bud burst and bud set are regulated by an epigenetic memory established by the temperature sum endured during embryogenesis. The resulting epitypes display a life-long shift in seasonal timing of the bud phenology, a trait previously presumed to be under strict classical selection and highly heritable. However, Norway spruce is a difficult plant to study because it has a very long generation time and an extensive genome size. We therefore seek to find a suitable perennial model plant to study the phenomenon of epigenetic climatic memory. Woodland strawberry (Fragaria vesca) may be an ideal model to research the role of epigenetic memory on plant phenology. Fragaria vesca is a perennial plant with a small well-characterized genome, a short sexual reproduction cycle and can also propagate asexually trough clonal daughter plants formed by stolons. We will explore whether the temperature sum experienced during sexual and asexual reproduction impact on the phenology of Fragaria vesca and use this as a model to decipher the molecular mechanism underlying epigenetic memory in plants.

Abstract

Bark beetles and their symbiotic bluestain fungi kill more trees than all other natural factors and cause great economic losses in Norway spruce and other conifers. The tree's natural defenses are the most important factor maintaining bark beetle-fungus complexes at low, endemic levels. Spraying Norway spruce trees with the plant hormone methyl jasmonate (MeJA) primes tree defenses without eliciting notable induced defenses, but enables the trees to respond much more quickly and strongly when challenged by bark beetles or fungi several weeks after treatment. This phenomenon, known as defense priming, is a form of acquired resistance that enables cost-effective and vigorous defense responses. In field experiments with 50-year-old clonal spruce trees terpene concentrations in the bark increased 60-fold within 24 h after mechanical wounding of MeJA primed trees, compared with a 13-fold increase in unprimed control trees. We also observed altered transcriptional patterns in primed trees using Illumina deep transcriptome sequencing. When wounded, primed trees launched vigorous induced defenses with significant differential regulation of gene transcripts, such as those involved in phenylpropanoid synthesis leading to lignification. Resistance-like genes, such as the NB-LRR coding genes, are also more rapidly induced in primed than in unprimed trees. Transcriptome results from primed but unwounded trees indicate an alteration in the state of the chromatin, resembling changes associated with the activity of the epigenetic machinery creating long-lasting epigenetic marks. We do not know yet how long the primed state is activated in Norway spruce, but our data so far indicate that it may last for at least 3 years.

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Abstract

The bark beetle Ips typographus carries numerous fungi that could be assisting the beetle in colonizing live Norway spruce (Picea abies) trees. Phenolic defenses in spruce phloem are degraded by the beetle's major tree-killing fungus Endoconidiophora polonica, but it is unknown if other beetle associates can also catabolize these compounds. We compared the ability of five fungi commonly associated with I. typographus to degrade phenolic compounds in Norway spruce phloem. Grosmannia penicillata and Grosmannia europhioides were able to degrade stilbenes and flavonoids faster than E. polonica and grow on minimal growth medium with spruce bark constituents as the only nutrients. Furthermore, beetles avoided medium amended with phenolics but marginally preferred medium colonized by fungi. Taken together our results show that different bark beetle-associated fungi have complementary roles in degrading host metabolites and thus might improve this insect's persistence in well defended host tissues.

Abstract

Plants are exposed to various pathogens in their environment and have developed immune systems with multiple layers of defence to fight-back. However, often pathogens overcome the resistance barriers, infect the plants to cause the disease. Pathogens that cause diseases on economically important crop plants like strawberry incur huge losses to the agriculture industry. For example, The 2016 outbreak of strawberry grey mould (Botrytis cinerea) in Norway caused up to 95% crop losses. Outbreaks like this underline the importance of developing novel and sustainable tools to combat plant diseases, for example by increasing the plants’ natural disease resistance. Priming plant defences using chemical elicitors may be effective in providing the enhanced resistance against multiple pathogens. We have used β-aminobutyric acid (BABA) as a chemical priming agent to induce resistance in Fragaria vesca against Botrytis cinerea. Effects of BABA on disease progression and defence responses of Fragaria are being characterized using molecular tools like RNAseq, RT-PCR and ChIP. As priming chemicals may induce an epigenetic memory in treated plants, we also plan to study the histone methylation patterns in primed plants and the genes that are regulated. Our long-term aim is to understand the duration of the epigenetic memory and its cross-generational transmission to the progeny in Fragaria. Our results will help guide various crop protection strategies in addition to providing new insights to develop novel tools for plant disease management.

Abstract

1 The European spruce bark beetle Ips typographus is a damaging pest on spruce in Europe. Beetle interactions with tree species originating outside the natural range of the beetle are largely unknown and may be unpredictable because trees without a co-evolutionary history with the beetle may lack effective defences. 2 The terpenoid composition and breeding suitability for I. typographus of the historic host Norway spruce Picea abies were compared with two evolutionary naïve spruces of North American origin that are extensively planted in North-West Europe: Sitka spruce Picea sitchensis and Lutz spruce Picea glauca x lutzii. 3 The bark of all three species had a similar chemical composition and similar levels of total constitutive terpenoids, although Norway spruce had higher total induced terpenoid levels. 4 Beetles tunnelling in the three spruce species produced similar amounts of aggregation pheromone. Controlled breeding experiments showed that I. typographus could produce offspring in all three species, with a similar offspring length and weight across species. However, total offspring production was much lower in Sitka and Lutz spruce. 5 Overall, the results of the present study suggest that I. typographus will be able to colonize Sitka and Lutz spruce in European plantations and in native spruce forests in North America if introduced there.

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Abstract

Global warming will most likely lead to increased drought stress in forest trees. We wanted to describe the adaptive responses of fine roots and fungal hyphae, at different soil depths, in a Norway spruce stand to long-term drought stress induced by precipitation exclusion over two growing seasons. We used soil cores, minirhizotrons and nylon meshes to estimate growth, biomass and distribution of fine roots and fungal hyphae at different soil depths. In control plots fine roots proliferated in upper soil layers, whereas in drought plots there was no fine root growth in upper soil layers and roots mostly occupied deeper soil layers. Fungal hyphae followed the same pattern as fine roots, with the highest biomass in deeper soil layers in drought plots. We conclude that both fine roots and fungal hyphae respond to long-term drought stress by growing into deeper soil layers.

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Abstract

The purpose of this study is to increase the basic understanding of outbreak dynamics in order to improve the management of bark beetle outbreaks. The spruce bark beetle Ips typographus is a major disturbance agent of European forests and is the continent’s most economically and environmentally damaging bark beetle. Outbreaks of the spruce bark beetle are often triggered by large windfall episodes, and we have utilized a unique opportunity to study a Slovakian outbreak where little salvage logging was performed in some areas after a 2.5 million m3 storm-felling in 2004. Our analyses focused on the first five years after the windfall, and we used a combination of empirical data and simulation models to understand the spatial patterns of beetle-killed forest patches developing during the outbreak. The univoltine beetle population used an increasing proportion of the windfelled trees during the two first seasons after the storm, but from the third season onwards our comparisons of inter-patch distance distributions indicated a transition from beetle production largely in windfall areas to a self-sustaining outbreak with infestation patches developing independently of the windthrows. The size of new infestation patches formed after this transition was modeled as a function of beetle pressure, estimated by the proportion of a circle area surrounding new patches that was covered by infestation patches the previous year. Our model results of patch size distribution did not correspond well with the empirical data if patch formation was modeled as a pure dispersal–diffusion process. However, beetle aggregation on individual trees appears to be important for patch development, since good correspondence with empirical data was found when beetle aggregation was incorporated in the modeled dispersal process. The strength of correspondence between the beetle aggregation model and the empirical data varied with the density of aggregation trees in the modeled landscape, and reached a maximum of 83% for a density of three aggregation trees per infestation patch. Our results suggest that efficient removal of windfelled trees up until the start of the second summer after a major windfall is important to avoid a transition into a patch-driven bark beetle outbreak that is very difficult to manage. Our results also indicate that the outcome of a patch-driven outbreak is difficult to predict, since the development of new infestation patches is not a simple function of beetle pressure but is also affected by beetle behavior and local forest conditions.

Abstract

Increasing inter-continental trade with wood chips represents a challenge for phytosanitary authorities, as such trade may lead to pest introductions and invasions with huge impacts on forest ecosystems and economy. Predicting species invasions and their impacts in advance may be difficult, but improved information about potential invasive species ahead of any interceptions is an important precautionary step to reduce the probability of invasions. Here we identify bark- and wood-boring insects that have a potential to become invasive in northern Europe, and that may be introduced by import of deciduous wood chips from North America. The potentially most damaging species belong to the beetle genus Agrilus (Buprestidae), which includes the highly damaging emerald ash borer A. planipennis. We give a brief presentation of this and seven other Agrilus species or subspecies, and review factors of importance for the risk of establishment and potential economic and ecological impacts of these species. We also discuss one Scolytinae, Hylurgopinus rufipes. There are strong indications in the literature that some north European trees are highly susceptible to attack from the selected beetle species. We therefore conclude that because north European trees have not coevolved with these herbivores and thus may lack adequate defenses, most of the identified beetle species are likely to spread in “defense- and enemy-free space” if they are introduced to northern Europe, with considerable economic and ecological consequences.

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Abstract

Trees must respond to many environmental factors during their development, and light is one of the main stimuli regulating tree growth. Thinning of forest stands by selective tree removal is a common tool in forest management that increases light intensity. However, morphological and anatomical adaptations of individual shoots to the new environmental conditions created by thinning are still poorly understood. In this study, we evaluated shoot morphology (shoot length, needle number, projected leaf area) and anatomy (tracheid lumen area, tracheid number, tracheid dimensions, xylem area, potential hydraulic conductivity) in three Norway spruce (Picea abies/L./Karst.) families exposed to different thinning regimes. We compared shoot characteristics of upper-canopy (i.e. sun-exposed) and lower-canopy (i.e. shaded) current-year shoots in a control plot and a plot thinned to 50 % stand density the previous year. One tree per family was chosen in each treatment, and five shoots were taken per canopy position. We found that upper-canopy shoots in both plots had higher values than lower-canopy shoots for all studied parameters, except lumen roundness and tracheid frequency (i.e. tracheid number per xylem area). Thinning had little effect on shoot morphology and anatomy 1 year after thinning, except for small but significant changes in tracheid dimensions. Needles were more sensitive to altered light conditions, as projected leaf area of shoot, needle number and leaf hydraulic conductivity changed after thinning. Differences between upper- and lower-canopy shoots did not seem to be influenced by thinning and were almost the same in both plots. Our results suggest that lower-canopy shoots require several years to modify their morphology and anatomy to new light conditions following thinning. The slow light adaptation of the lower canopy may be of practical importance in forest management: thinned stands may be predisposed to drought stress because newly exposed shoots experience increased illumination and transpiration after thinning.

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Abstract

Bark beetles and associated fungi are among the greatest natural threats to conifers worldwide. Conifers have potent defenses, but resistance to beetles and fungal pathogens may be reduced if tree stored resources are consumed by fungi rather than used for tree defense. Here, we assessed the relationship between tree stored resources and resistance to Ceratocystis polonica , a phytopathogenic fungus vectored by the spruce bark beetle Ips typographus. We measured phloem and sapwood nitrogen, non-structural carbohydrates (NSC), and lipids before and after trees were attacked by I. typographus (vectoring C. polonica) or artificially inoculated with C. polonica alone. Tree resistance was assessed by measuring phloem lesions and the proportion of necrotic phloem around the tree’s circumference following attack or inoculation. While initial resource concentrations were unrelated to tree resistance to C. polonica, over time, phloem NSC and sapwood lipids declined in the trees inoculated with C. polonica. Greater resource declines correlated with less resistant trees (trees with larger lesions or more necrotic phloem), suggesting that resource depletion may be caused by fungal consumption rather than tree resistance. Ips typographus may then benefit indirectly from reduced tree defenses caused by fungal resource uptake. Our research on tree stored resources represents a novel way of understanding bark beetle-fungal-conifer interactions.

Abstract

Insects are among the organisms that will react most rapidly to climate change. They have a short generation span, they are very mobile, and their rate of development is directly impacted by temperature. This means that the development from egg to adult insect is much quicker when temperatures rise – a fact that may bode bad news for forestry.

Abstract

This chapter provides an overview of anatomical and ecological aspects of resin-based defences in pines and contrasts the defence strategy of pines with that of other conifers. The main constituents of conifer resin are mono- and diterpenes in about equal amounts, with smaller amounts of sesquiterpenes. Resin production and storage represent a great cost for the trees, and because resin is both chemically toxic and physically deterring to insects and pathogens it has long been considered an important defence mechanism in conifers. Preformed or constitutive resin structures are present in pines and all other members of the pine family, but are generally absent in non-Pinaceae species. Resin stored under pressure in constitutive ducts flows out when a tree is injured and helps trapping or repelling invading organisms and sealing the wound. Pines have constitutive resin ducts in needles, phloem and xylem. In the phloem and xylem constitutive resin ducts are oriented both radially (within the radial rays) and axially in the form of cortical resin ducts in the outer phloem and constitutive resin ducts in the xylem. Numerous connections between the radial resin ducts and the axial resin ducts in the xylem create a large inter-connected resin reservoir. In addition, so-called traumatic resin ducts can be induced axially in the xylem in response to wounding, insect attack or other biotic and abiotic stresses. Traumatic resin ducts may contribute to so-called acquired or systemic induced resistance that increases tree resistance to future attacks.

Abstract

We compared gene expression in Norway spruce secondary phloem (bark) and developing xylem (sapwood) in response to the necrotrophic pathogen Heterobasidion parviporum, wounding and methyl jasmonate (MeJ). The pathogen induced systemic and local up-regulation of PaPX3, PaPX2 and PaChi4 in both bark and sapwood that returned to constitutive levels as the plants recovered from the infection, whereas the local responses to MeJ were similar in both tissues but was longer lasting for PaPX3 and PaChi4. Genes involved in lignin biosynthesis (PaPAL1, PaPAL2, PaC4H3/5 and PaHCT1) were up-regulated locally in the bark in response to pathogen and wounding whereas MeJ induced a similar but stronger local response. The ethylene biosynthesis related transcripts PaACO and PaACS did not increase in response to MeJ treatment or the pathogen, however it increased both locally and systemically as a response to wounding in the sapwood. These results demonstrate that the local and systemic host responses to pathogen infection and wounding largely correspond and reveal striking similarities between the local response to a necrotroph, wounding and MeJ treatment in both bark and living wood.

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Abstract

Purpose: Drought-induced tree susceptibility is a major risk associated with climate change. Here we report how an 11-week drought affected tracheid structure, gene expression, and above- and belowground growth in 5-year-old Norway spruce trees (Picea abies) under controlled conditions. Results: The canopy of trees subjected to severe drought had significantly less current-year needle biomass, and fewer tracheids and tracheid rows in current-year shoots compared to fully watered control trees. Belowground tissues were more strongly affected by drought than aboveground tissues. In fine roots (<2 mm diameter) severe drought significantly reduced root biomass, root diameter, root length density and root surface area per soil volume compared to the control. Tracheid diameter and hydraulic conductivity in fine roots were significantly lower and tracheid flatness higher in trees subjected to severe drought than in control trees, both for long and short roots. Transcripts of the drought-related dehydrins PaDhn1 and PaDhn6 were strongly upregulated in stem bark and current-year needles in response to drought, whereas PaDhn4.5 was down-regulated. Conclusions: This study demonstrates that drought reduces biomass and hydraulic conductivity in fine roots and needles. We suggest that the ratio between PaDhn6 and PaDhn4.5 may be a sensitive marker of drought stress in Norway spruce.

Abstract

Ceratocystis polonica and Heterobasidion parviporum are important fungal pathogens in Norway spruce (Picea abies). Tree susceptibility to these pathogens with respect to phenology was studied using artificial fungal inoculations at six stages of bud development, and assessed by measuring phloem necroses in the stems of 2- and 8-year-old trees. Tree capacity for resistance was assessed by measuring phloem nonstructural carbohydrates at each stage. Phloem necroses were significantly larger in trees with fungal versus control inoculations and increased significantly over time. Changes in nonstructural carbohydrates occurred in the trees; a significant decline in starch and a slight but significant increase in total sugars occurred over time. These results suggest that susceptibility to fungal pathogens and carbohydrate levels in the stems of the trees were related to fine-scale changes in bud development. A trade-off may occur between allocation of starch (the major fraction of the stem carbohydrate pool) to bud development/shoot growth versus defence of the stem. Previous tests of plant defence hypotheses have focused on herbivory on plants growing under different environmental conditions, but the role of phenology and the effect of pathogens are also important to consider in understanding plant resource allocation patterns.

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Abstract

Conifer needles are extraordinarily variable and much of this diversity is linked to the water transport capacity of the xylem and to xylem conduit properties. However, we still know little about how anatomical characteristics influence the hydraulic efficiency of needle xylem in different parts of the crown. In this study we evaluated needle function and anatomy in Norway spruce families exposed to different light conditions. We measured tracheid and needle characteristics of sun-exposed and shaded current-year needles in two experimental plots: a control plot and a thinned plot with 50% reduction in stand density. Sun-exposed needles had a larger tracheid lumen area than shaded needles, and this was caused by a larger maximum tracheid lumen diameter, while the minimum lumen diameter was less plastic. Sun-exposed needles had also higher theoretical hydraulic conductivity than shaded needles. Thinning leads to increased radiation to the lower branches, and presumably exposes the upper branches to stronger water stress than before thinning. Thinning affected several needle parameters both in sun-exposed and shaded needles; tracheid lumens were more circular and minimum tracheid lumen diameter was larger in the thinned plot, whereas maximum tracheid lumen diameter was less plastic on both plots. This study demonstrates that needle xylem structure in Norway spruce is clearly influenced by the light gradient within the tree crown.

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Abstract

Secondary attraction to aggregation pheromones plays a central role in the host colonization behavior of the European spruce bark beetle Ips typographus. However, it is largely unknown how the beetles pioneering an attack locate suitable host trees, and eventually accept or reject them. To find possible biomarkers for host choice by I. typographus, we analyzed the chemistry of 58 Norway spruce (Picea abies) trees that were subsequently either (1) successfully attacked and killed, (2) unsuccessfully attacked, or (3) left unattacked. The trees were sampled before the main beetle flight in a natural Norway spruce-dominated forest. No pheromones were used to attract beetles to the experimental trees. To test the trees’ defense potential, each tree was treated in a local area with the defense hormone methyl jasmonate (MeJ), and treated and untreated bark were analyzed for 66 different compounds, including terpenes, phenolics and alkaloids. The chemistry of MeJ-treated bark correlated strongly with the success of I. typographus attack, revealing major chemical differences between killed trees and unsuccessfully attacked trees. Surviving trees produced significantly higher amounts of most of the 39 analyzed mono-, sesqui-, and diterpenes and of 4 of 20 phenolics. Alkaloids showed no clear pattern. Differences in untreated bark were less pronounced, where only 1,8-cineole and (−)-limonene were significantly higher in unsuccessfully attacked trees. Our results show that the potential of individual P. abies trees for inducing defense compounds upon I. typographus attack may partly determine tree resistance to this bark beetle by inhibiting its mass attack.

Abstract

Background: NB-LRR resistance proteins are involved in recognizing pathogens and other exogenous stressors in plants. Resistance proteins are the first step in induced defence responses and a better understanding of their regulation is important to understand the mechanisms of plant defence. Much of the post-transcriptional regulation in plants is controlled by microRNAs (miRNA). We examined the expression of five Norway spruce miRNA that may regulate NB-LRR related transcripts in secondary phloem (bark) of resistant Norway spruce after wounding and inoculation with the necrotrophic blue stain fungus Ceratocystis polonica. Results: The plants of this clone recovered from both the pathogen inoculations and wounding alone. We found local and systemic induction of the resistance marker genes PaChi4, PaPAL and PaPX3 indicative of an effective induced host defence response. There were minor local and systemic changes in the expression of five miRNAs and 21 NB-LRRs between healthy and treated plants. Only five putative NB-LRRs (PaLRR1, PaLRR3, PaLRR14, PaLRR15 and PaLRR16) showed significant increases greater than two-fold as a local response to C. polonica. Of all NB-LRRs only PaLRR3, the most highly differentially regulated NB-LRR, showed a significant increase also due to wounding. The five miRNAs showed indications of an initial local and systemic down-regulation at day 1, followed by a later increase up to and beyond the constitutive levels at day 6. However, the initial down-regulation was significant only for miR3693 and miR3705. Conclusions: Overall, local and systemic expression changes were evident only for the established resistance marker genes and PaLRR3. The minor expression changes observed both for the followed miRNAs and their predicted NB-LRR targets suggest that the expression of most NB-LRR genes are maintained close to their constitutive levels in stressed and healthy Norway spruce plants.

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Abstract

Norway spruce (Picea abies) bark contains specialized phloem parenchyma cells that swell and change their contents upon attack by the bark beetle Ips typographus and its microbial associate, the blue stain fungus Ceratocystis polonica. These cells exhibit bright autofluorescence after treatment with standard aldehyde fixatives, and so have been postulated to contain phenolic compounds. Laser microdissection of spruce bark sections combined with cryogenic NMR spectroscopy demonstrated significantly higher concentrations of the stilbene glucoside astringin in phloem parenchyma cells than in adjacent sieve cells. After infection by C. polonica, the flavonoid (+)-catechin also appeared in phloem parenchyma cells and there was a decrease in astringin content compared to cells from uninfected trees. Analysis of whole-bark extracts confirmed the results obtained from the cell extracts and revealed a significant increase in dimeric stilbene glucosides, both astringin and isorhapontin derivatives (piceasides A to H), in fungus-infected versus uninfected bark that might explain the reduction in stilbene monomers. Phloem parenchyma cells thus appear to be a principal site of phenolic accumulation in spruce bark.

Abstract

1. Neodiprion sertifer nucleopolyhedrovirus (NeseNPV) is widely used as a viral bio-insecticide against larvae of the European pine sawfly N. sertifer (Geoff.) (Hymenoptera: Diprionidae), which is one of the most harmful defoliators of pines in Northern Europe. A major obstacle to studying this pathogenic virus in nature is the difficulty of confirming and quantifying the presence of NeseNPV. 2. In the present study, we developed real-time polymerase chain reaction (PCR) primers, based on the caspid gene 39 sequence, for the specific and quantitative detection of NeseNPV. The quantitative real-time PCR (qPCR) assay can detect virus from any substrate tested, including different insect life stages (egg, larval, adult), pine foliage, and litter or ground vegetation. The reproducible detection limit for the real-time assay is 0.013 pg of viral DNA (0.013 × 10−12 g), corresponding to 136 viral genomes or approximately one to seven virus occlusion bodies per sample. 3. qPCR is a specific, quantitative, sensitive, reliable and flexible procedure, and is a good supplement to conventional microscopy- or bioassay-based methods for detection of the virus. We have used qPCR to quantify the level of NeseNPV in samples collected in the field after aerial application of the virus, and demonstrated significantly higher virus levels in sawfly larvae from sprayed areas compared with unsprayed control areas 4 weeks after spraying. 4. This qPCR assay can be used to determine important aspects of the biology of NeseNPV (e.g. virus levels in different insect life stages and in their microhabitats on pine foliage and in forest litter).

Abstract

Damage by cone and seed insects is a severe impediment to those producing conifer seeds in seed orchards. The SNS-supported network `Nordic cone and seed insects` has been cooperating to address this problem since 2007. One of the outcomes is the coordinated monitoring of the most important insect species. The data collected form a basis for establishing strategies for pest management.

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Abstract

Conifers and other trees are constantly adapting to changes in light conditions, water/nutrient supply and temperatures by physiological and morphological modifications of their foliage. However, the relationship between physiological processes and anatomical characteristics of foliage has been little explored in trees. In this study we evaluated needle structure and function in Norway spruce families exposed to different light conditions and transpiration regimes. We compared needle characteristics of sun-exposed and shaded current-year needles in a control plot and a thinned plot with 50% reduction in stand density. Whole-tree transpiration rates remained similar across plots, but increased transpiration of lower branches after thinning implies that sun-exposed needles in the thinned plot were subjected to higher water stress than sun-exposed needles in the control plot. In general, morphological and anatomical needle parameters increased with increasing tree height and light intensity. Needle width, needle cross-section area, needle stele area and needle flatness (the ratio of needle thickness to needle width) differed most between the upper and lower canopy. The parameters that were most sensitive to the altered needle water status of the upper canopy after thinning were needle thickness, needle flatness and percentage of stele area in needle area. These results show that studies comparing needle structure or function between tree species should consider not only tree height and light gradients, but also needle water status. Unaccounted for differences in needle water status may have contributed to the variable relationship between needle structure and irradiance that has been observed among conifers.

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Abstract

We treated Norway spruce (Picea abies) stems with methyl jasmonate (MeJA) to determine possible quantitative and qualitative eVects of induced tree defenses on pheromone emission by the spruce bark beetle Ips typographus. We measured the amounts of 2-methyl-3-buten-2-ol and (S)-cis-verbenol, the two main components of the beetle’s aggregation pheromone, released from beetle entrance holes, along with phloem terpene content and beetle performance in MeJA-treated and untreated Norway spruce logs. As expected, phloem terpene levels were higher and beetle tunnel length was shorter (an indication of poor performance) in MeJA-treated logs relative to untreated logs. Parallel to the higher phloem terpene content and poorer beetle performance, beetles in MeJA-treated logs released signiWcantly less 2-methyl-3-buten-2-ol and (S)-cis-verbenol, and the ratio between the two pheromone components was signiWcantly altered. These results suggest that host resistance elicited by MeJA application reduces pheromone emission by I. typographus and alters the critical ratio between the two main pheromone components needed to elicit aggregation. The results also provide a mechanistic explanation for the reduced performance and attractivity observed in earlier studies when bark beetles colonize trees with elicited host defenses, and extend our understanding of the ecological functions of conifer resistance against bark beetles.

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Abstract

Background: Tree-killing bark beetles (Coleoptera, Scolytinae) are among the most economically and ecologically important forest pests in the northern hemisphere. Induction of terpenoid-based oleoresin has long been considered important in conifer defense against bark beetles, but it has been difficult to demonstrate a direct correlation between terpene levels and resistance to bark beetle colonization. Methods: To test for inhibitory effects of induced terpenes on colonization by the spruce bark beetle Ips typographus (L.) we inoculated 20 mature Norway spruce Picea abies (L.) Karsten trees with a virulent fungus associated with the beetle, Ceratocystis polonica (Siem.) C. Moreau, and investigated induced terpene levels and beetle colonization in the bark. Results: Fungal inoculation induced very strong and highly variable terpene accumulation 35 days after inoculation. Trees with high induced terpene levels (n = 7) had only 4.9% as many beetle attacks (5.1 vs. 103.5 attacks m22) and 2.6% as much gallery length (0.029 m m22 vs. 1.11 m m22) as trees with low terpene levels (n = 6). There was a highly significant rank correlation between terpene levels at day 35 and beetle colonization in individual trees. The relationship between induced terpene levels and beetle colonization was not linear but thresholded: above a low threshold concentration of ,100 mg terpene g21 dry phloem trees suffered only moderate beetle colonization, and above a high threshold of ,200 mg terpene g21 dry phloem trees were virtually unattacked. Conclusion/Significance: This is the first study demonstrating a dose-dependent relationship between induced terpenes and tree resistance to bark beetle colonization under field conditions, indicating that terpene induction may be instrumental in tree resistance. This knowledge could be useful for developing management strategies that decrease the impact of tree-killing bark beetles.

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Abstract

Norway spruce (Picea abies) defends itself against herbivores and pathogens by formation of traumatic resin ducts filled with terpenoid-based oleoresin. An important group of enzymes in terpenoid biosynthesis are the short-chain isoprenyl diphosphate synthases which produce geranyl diphosphate (C10), farnesyl diphosphate (C15), and geranylgeranyl diphosphate (C20) as precursors of monoterpenes, sesquiterpenes, and diterpene resin acids, respectively. After treatment with methyl jasmonate (MJ) we investigated the expression of all isoprenyl diphosphate synthase genes characterized to date from Norway spruce and correlated this with formation of traumatic resin ducts and terpene accumulation. Formation of traumatic resin ducts correlated with higher amounts of monoterpenes, sesquiterpenes and diterpene resin acids and an upregulation of isoprenyl diphosphate synthase genes producing geranyl diphosphate or geranylgeranyl diphosphate. Among defense hormones, jasmonate and jasmonate-isoleucine conjugate accumulated to higher levels in trees with extensive traumatic resin duct formation, whereas salicylate did not. Jasmonate and ethylene are likely to both be involved in formation of traumatic resin ducts based on elevated transcripts of genes encoding lipoxygenase and 1-aminocyclopropane-1-carboxylic acid oxidase associated with resin duct formation. Other genes involved in defense signalling in other systems, mitogen-activated protein kinase3 and nonexpressor of pathogenesis-related gene1, were also associated with traumatic resin duct formation. These responses were detected not only at the site of MJ treatment, but also systemically up to 60 cm above the site of treatment on the trunk.

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Abstract

The Eurasian spruce bark beetle, Ips typographus, is one of the major forest insect pests in Europe, capable of mass-attacking and killing mature Norway spruce trees. The initiation and development of a new generation are strongly controlled by temperature and a warmer climate may affect the number of generations that is produced per year and hence the outbreak dynamics. Experimental knowledge regarding reproductive diapause adaptations is, however, too sparse for largescale assessments of future trends. We developed a model description of diapause induction, and used gridded observational temperature data to evaluate multiple combinations of day length and temperature thresholds to find the model parameterisation most coherent with I. typographus monitoring data from Scandinavia. The selected model parameterisation is supported by European literature data, though further experimental studies are required to analyse population specific adaptations and capacity for adjustments to changing climate conditions. Implementing the model description of reproductive diapause in a temperature driven model of bark beetle phenology (swarming activity and development from egg to mature bark beetle), enabled us to assess the length of the late summer swarming period that is a critical determinant of the risk of forest damage. By using regional climate model data we show that higher temperatures can result in increased frequency and length of late summer swarming events, producing a second generation in southern Scandinavia and a third generation in lowland parts of central Europe. Reproductive diapause will not prevent the occurrence of an additional generation per year, but the day length cues may restrict the length of the late summer swarming period.

Abstract

Today the spruce bark beetle Ips typographus is always univoltine in Northern Europe including Norway and completes development from egg to adult between May and August. Further south in Europe, development is bivoltine with the completion of two generations in most years. A temperature-driven developmental model suggests that by 2070-2100 the voltinism of I. typographus will change dramatically in Norway. If summers become only 2.5°C warmer than today bivoltinism can be expected every single year in the major spruce growing areas in S-Norway. This is likely to have dramatic effects on forestry since two generations per year will give two, instead of one, attack periods each summer. In addition to increasing the number of attacked trees the effect of the attacks may also be more severe, as Norway spruce is more susceptible to beetle attacks later in the summer. However, climate change will probably also change the phenology of Norway spruce and thus its susceptibility to attack by I. typographus and its phytopathogenic fungal associates. We are currently modelling how tree resistance varies with temperature and tree phenology in order to provide more well-founded advice to forest managers on the interaction between bark beetles and tree in a future climate.

Abstract

We monitored the effects of the drought stress on 20-year old clones of Norway spruce (Picea abies) by using a range of instrumental methods. On two experimental plots (Hoxmark, Norway, 59°40\"14`N, 10°47\"36`E) the drought was induced in a period between May and October 2009 by removing the throughfall using the rain shelters and trenching. We collected data on soil moisture, stem and branch sap flow, xylem diameter, anatomical and calorimetric analysis of the needles, fine root biomass and dynamics and resistance to pathogens. Standard meteorological data were collected locally throughout the whole period. Here we present the preliminary analysis of sap flow and xylem diameter in a period 1-17 august 2009. The sap flow was measured on stems in the breast height by using the method of stem tissue heat balance (THB, EMS Brno). The values were measured once in 2 minutes and saved as the average of 10 minutes. The fluctuations in xylem diameter was monitored by using the automatic dendrometers DR26 (EMS Brno). We evaluated both diurnal and seasonal dynamics. Preliminary results show a significant difference in shape of diurnal curves of transpiration as well as different time lag among the sap flow and the potential evapo-transpiration. Also the differences in diurnal dynamics of the stem circumference suggested different xylem water potential in stressed and control trees. In the drought-stressed trees the diurnal fluctuation in stem diameter was about 4 times higher and the total stem increment one third lower, com-pared to the control trees.

Abstract

Cone and seed insects reduce seed production in seed orchards by feeding on cones and/or seeds and thus cause a lack of seedlings for reforestation. One of the most serious pest species in conifers is the spruce cone worm, Dioryctria abietella. Since 2007, we have used pheromone traps to monitor the flight of this species in Denmark, Finland, Norway and Sweden (and Estonia from 2009). In addition to monitoring, all countries have measured air temperatures in the vicinity of the traps. It is thus possible to correlate flight activity with temperature and temperature sums (accumulated day degrees >5°C). In Sweden cone development has also been registered throughout the monitoring period. Our results show that D. abietella has an extended flight period in Northern Europe, which lasts from late May to late September. Further research is needed to determine if late flying individuals oviposit on shoots or in cones. Increased knowledge about the flight period of D. abietella and how it coincides with temperature and cone development may help us develop better control measures against this important pest.

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Abstract

The blue-stain fungus Ceratocystis resinifera colonizes wounds on living Picea spp. and other conifers in Europe and North America. Little is known regarding the pathogenicity of this fungus and consequently, four Norwegian C. resinifera isolates were inoculated on to Norway spruce (Picea abies) using two different techniques. These included single-point inoculations on young trees (two inoculations per tree on 14-year-old trees) and mass-inoculations on older trees (∼200 inoculations per tree on 34-year-old trees). In both experiments, C. resinifera induced minor symptoms that in most cases did not differ significantly from inoculation with sterile agar. The virulent blue-stain fungus C. polonica, which was inoculated for comparative purposes, induced extensive symptoms, causing 83% dead cambium circumference and 82% blue-stained sapwood, and long necrotic lesions in the phloem. The results suggest that C. resinifera is non-pathogenic or only mildly pathogenic to Norway spruce and does not present a threat to these trees.

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Abstract

Constitutive and inducible terpene production is involved in conifer resistance against bark beetles and their associated fungi. In this study 72 Norway spruce (Picea abies) were randomly assigned to methyl jasmonate (MJ) application, inoculation with the bluestain fungus Ceratocystis polonica, or no-treatment control. We investigated terpene levels in the stem bark of the trees before treatment, 30 days and one year after treatment using GC–MS and two-dimensional GC (2D-GC) with a chiral column, and monitored landing and attack rates of the spruce bark beetle, Ips typographus, on the trees by sticky traps and visual inspection. Thirty days after fungal inoculation the absolute amount and relative proportion of (+)-3-carene, sabinene, and terpinolene increased and (+)-α-pinene decreased. Spraying the stems with MJ tended to generally increase the concentration of most major terpenes with minor alteration to their relative proportions, but significant increases were only observed for (−)-β-pinene and (−)-limonene. Fungal inoculation significantly increased the enantiomeric ratio of (−)-α-pinene and (−)-limonene 1 month after treatment, whereas MJ only increased that of (−)-limonene. One year after treatment, both MJ and fungal inoculation increased the concentration of most terpenes relative to undisturbed control trees, with significant changes in (−)-β-pinene, (−)-β-phellandrene and some other compounds. Terpene levels did not change in untreated stem sections after treatment, and chemical induction by MJ and C. polonica thus seemed to be restricted to the treated stem section. The enantiomeric ratio of (−)-α-pinene was significantly higher and the relative proportions of (−)-limonene were significantly lower in trees that were attractive to bark beetles compared to unattractive trees. One month after fungal inoculation, the total amount of diterpenes was significantly higher in putative resistant trees with shorter lesion lengths than in putative susceptible trees with longer lesions. Thus, terpene composition in the stem bark may be related to resistance of Norway spruce against I. typographus and C. polonica.

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Abstract

The anamorph genus Leptographium Lagerberg and Melin includes species that are typically bark beetleassociated fungi, with teleomorphs in Grosmannia. During a survey of ophiostomatoid fungi in Norway, two unusual species, that fit the broader morphological description of Leptographium, were isolated directly from the rootfeeding beetles, Dryocetes authographus and Hylastes cunicularius, as well as from roots infested by these insects. The first of these could be distinguished from other described species based on a sparse sporulation, black spore drops and chlamydospores in older cultures. This species also produces a Hyalorhinocladiella synanamorph. The second species was characterised by distinctly curved conidia. Based on these unusual morphological characteristics and distinct DNA sequences, these fungi were recognised as new taxa for which the names Leptographium chlamydatum sp. nov. and L. curvisporum sp. nov. are provided.

Abstract

The interaction between conifers, ophiostomatoid fungi and bark beetles is a key factor in conifer ecosystems worldwide, since combined beetle-fungus attacks may cause massive tree mortality, huge economical losses and landscape-level ecological changes. For more than a century researchers have been arguing about who is responsible for tree mortality in bark beetle attacked trees - the beetles themselves or their associated fungi. As in many such polarized debates the best answer probably lies somewhere in between the extremes. The beetles are obviously central in tree killing, as they are actively selecting suitable host trees, short-circuiting tree defenses by boring straight into the relatively defenseless cambial area, and causing mechanical damage to the phloem. However, ophiostomatoid fungi are equally obvious contributors to tree death since nearly all tree-killing bark beetles are associated with such fungi, these fungi are able to colonize and kill healthy phloem and sapwood far beyond the beetle tunnels, and many fungi can kill healthy trees in experimental mass-inoculations. Thus, the trees are facing a beetle-fungus complex that probably acts synergistically to overwhelm tree defenses. The beetles’ mass-attack strategy is central to the success of the beetle-fungus complex, since multiple attacks deplete tree defenses and speed up tree-killing.

Abstract

Clonal variation towards resistance has been observed in Norway spruce Heterobasidion annosum s.l. (H.a). H.a. is the main cause of root rot and has a severe economic impact on an economically important conifer tree species. Annual financial losses are in the hundreds of millions of Euros annually. Less susceptible clones appear to have an efficient system of recognizing the pathogen and initiating early defense signalling events. Active defense responses can be started locally and transmitted systemically. This work focus on the expression both spatially (systemically) and temporally in this pathosystem. Two-year-old, somatic saplings of the Norway spruce clone were challenged with H.a., wounded, methyl jasmonate painted and compared to untreated controls and ninety plants were used for the experiment. Stem samples were collected at 1, 3, 6 and 13 days post inoculation (d.p.i). The stem of the saplings were divided into sections along its length and the bark and wood separated from each other at time of collection. In order to see local response an area of 1cm including the site of inoculation was collected, while the spatial (systemic) response was assessed in sections collected at distances of 3 and 6cm away from the site of inoculation. The separated bark and wood were analysed for differential gene expression by qRT-PCR, and the results from peroxidases (PaPX3 and PaPX2) and a chitinase (PaChi4) are presented. Both local and systemic up- and down-regulation were observed at the transcriptional level in both bark and wood, up to 2000 fold local increase in expression was observed for PaChi4.

Abstract

Combined bark beetle-fungus attacks are a key factor in conifer ecosystems in the Northern Hemisphere, where they regularly cause massive tree mortality. Central to the success of the bark beetle-fungus complex is the fact that bark beetles are among the few insects that are capable of breaching the potent anatomical and chemical defenses of healthy tree stems.....

Abstract

Tree-killing bark beetles conquer healthy trees by means of pheromone mediated mass-attacks. The initial attackers select trees through a process of random landing and processing of tactile and short-range chemical stimuli. Inside acceptable hosts they produce aggregation pheromones that attract additional attackers. In a recent field experiment we induced defense responses in 60-year-old Norway spruce trees and monitored colonization by the spruce bark beetle Ips typographus. Induced stem sections had significantly less beetle colonization than control sections, with shorter parental galleries and fewer eggs deposited. In addition, fewer beetles were attracted to beetles tunnelling in induced bark. This reduced ability to attract conspecifics suggests that induced host defenses interfere with beetle pheromone production or emission. The mechanisms responsible for this are unknown, but may involve emission of volatile terpenes such as (E)-b-farnesene and linalool, which have been shown to increase up to 100-fold in induced Norway spruce trees. These compounds have been reported to attract natural enemies of herbivores or repel herbivores directly in other conifer-insect interactions.

Abstract

Seed production in Norway spruce (Picea abies) seed orchards may be severely reduced by insects attacking cones and seeds. Insects in several orders and genera are responsible for the damage, and at present there are no effective and environmentally acceptable control options. Studies in Sweden have shown promising results with spraying Bt (Bacillus thuringiensis var. kurstaki x aizawai) against species of Lepidoptera, but for other pests other control methods must be developed....

Abstract

The European pine sawfly Neodiprion sertifer is a widely distributed defoliator of pines that can cause substantial growth losses over extensive areas. It attacks most species of twoneedle pines in its distribution area, and have occasional short-lived outbreaks that may cover up to 200.000 ha. In Norway we have had outbreak populations in the eastern part of the country since 2004, and in an ongoing research project we are evaluating aerial application of the Neodiprion sertifer nuclear polyhedrosis virus (NsNPV) to control Neodiprion sertifer....

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Abstract

Host tree terpenes can influence attraction of conifer-infesting bark beetles to their aggregation pheromones, and both synergistic and inhibitory effects have been reported. We tested a gradient of ratios of (–)-α-pinene, the predominant monoterpene in Norway spruce, to the pheromone of Ips typographus, a major pest of Norway spruce. Attraction of I. typographus increased as the release rate of (–)-α-pinene increased. The two highest (–)-α-pinene : pheromone ratios (526 : 1 and 2595 : 1) attracted twice as many I. typographus as pheromone alone, whereas low to intermediate ratios (56 : 1, 274 : 1) did not differ from pheromone alone. Our results are in agreement with a proposed model, which suggests that bark beetles display unique response profiles to host terpenes depending on the physiological condition of the host trees that they typically colonize. Ips typographus, which is an aggressive species capable of colonizing and killing healthy trees, showed an increased attraction to monoterpene : pheromone ratios, and this may be high enough to inhibit attraction of less aggressive beetle species typically colonizing dead, dying or stressed trees. Attraction of associates of I. typographus was also modified by (–)-α-pinene. Ips duplicatus, a competitor of I. typographus, showed increased attraction to the pheromone of I. typographus across all concentrations of (–)-α-pinene.

Abstract

The Norwegian Monitoring Programme for Forest Damage (OPS) has since its start registered damage to selected trees. The aim of the registrations has been to explain variations in crown density and crown colour. In answer to international requests, the Norwegian Forest and Landscape Institute has prepared a short guide to the determination of the most common forms of damage found in Norwegian forests...

Abstract

Model simulations show that an increased frequency in storms and drought periods may result in more frequent and shorter outbreaks of bark beetles. Warmer summers can result in two bark beetle generations per summer instead of one, giving bark beetles the opportunity to attack forests twice in a single year.

Abstract

Studies of defense mechanisms in Norway spruce Picea abies against the spruce bark beetle Ips typographus show that inducible defenses are triggered by unsuccessful beetle attacks. Moreover, spruce trees can be vaccinated against bark beetle attacks through stem bark inoculation with Ceratocystis polonica, an aggressive fungal associate of Ips typographus. Artificial inoculation with certain other fungi has a similar effect, making treated trees a tougher mach for attacking beetles. A similar enhancement of resistance can be achieved by stem surface treatment with methyl jasmonate. Enhanced resistance is associated with the formation of traumatic resin ducts, and mobilization of phenolic substances in living parenchyma cells. The phenolics appear to mix with the traumatic resin. Similar mechanisms are operating in other conifers. We hypothesize that unsuccessful beetle attacks produce a vaccination effect in the trees, enhancing their resistance to later attack. Adding the effect of induced resistance as a model factor shows an immediate effect on the population dynamics of Ips typographus.

Abstract

Bark beetles and associated phytopathogenic fungi elicit defence responses in conifers that may interfere with beetle establishment and development. Norway spruce is serving as a useful model species for studies of induced defences elicited by beetle attacks, fungal inoculation, and treatment with chemical elicitors.When trees are pretreated with a sublethal dose of fungal inoculations or with the phytohormone methyl jasmonate they become much more resistant to subsequent bark beetle attacks or artificial mass inoculations with fungi. This induced disease resistance follows dose-response dynamics, is nonspecific with respect to the pretreatment organism, appears to be nonsystemic, takes weeks rather than days to become activated, and can also be activated by mechanical wounding alone.Application of methyl jasmonate to Norway spruce stems induces a massive increase in terpene levels and external resin flow on the stem, whereas no increase is observed in soluble phenolics. Methyl jasmonate-application also leads to significantly less bark beetle colonization, with shorter parental galleries and fewer eggs laid in treated bark. There were also reductions in the number of beetles produced and the mean dry weight per beetle in methyl jasmonate-treated bark. Furthermore, fewer beetles were attracted to conspecifics tunneling in MJ-treated bark.The exact mechanisms responsible for induced resistance in Norway spruce and other conifers have not been determined, but inducible anatomical defense responses such as changes in polyphenol-containing parenchyma cells (PP cells) in the phloem and induction of traumatic resin duct formation in the sapwood seem to play an important role.

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Abstract

When conifers such as Picea abies Karst. (Norway spruce) are attacked by insects or pathogens, they often respond by producing increased quantities of terpenoid oleoresin. This response can be mimicked in young P. abies seedlings by treatment with methyl jasmonate (MJ). In this study, we determined the effects of MJ on terpenoids and other chemical defenses of mature P. abies, and investigated whether this treatment protected trees against attack by the blue-stain fungus Ceratocystis polonica (Siem.) C. Moreau, the most important fungal associate of the spruce bark beetle Ips typo-graphus L. Methyljasmonate treatment induced the formation of traumatic resin ducts in the developing xylem, enhanced resin flow and stimulated increased accumulation of monoterpenes, sesquiterpenes and diterpene resin acids. However, only minor changes were detected in terpene composition in response to MJ treatment and no changes in soluble phenolic concentration were measured. There was much variability in the timing and degree of response to MJ among clones. The observed chemical and anatomical changes in response to MJ treatment were correlated with increased resistance to C. polonica, suggesting that terpenoid oleoresin may function in defense against this pathogen.

Abstract

The spruce bark beetle is the most serious pest of mature spruce trees in Eurasia. At low population densities it breeds in weakened or newly dead trees, but at high densities it becomes a tree killer. The exact factors triggering outbreaks are not fully understood, but climatic variables are important candidates. Populations in SE Norway have been monitored since 1979. Various time series methods are used to explore the role of climate in outbreaks of the spruce bark beetle in space and time, and to estimate the distribution of bivoltism under different scenarios of climate change. Large windfall events appear to be a major synchronizer of beetle outbreaks in areas subjected to regionalized weather systems, and the northern border of bivoltism may be moved towards north as a function of increasing temperature. Preliminary models of the population dynamics emphasize the frequency of large windfall events and phenological changes due to temperature increase. Final aims are to estimate the regional risks of forest damage under different scenarios of climate change, and to describe practices that may reduce the impact for forest managers.

Abstract

At least three mechanisms are known to cause synchrony among spatially separated insect populations: 1) dispersal among populations, 2) synchronous stochastic effects, often referred to as the Moran effect, and 3) trophic interactions with other species that are either themselves synchronized or mobile.The present study brings in the role of insect taxa for spatial synchrony. The spatial synchrony observed in several North American and Eurasian epidemic bark beetles was compared with patterns of synchrony in outbreaks of defoliating forest Lepidoptera, revealing a marked difference between these two major insect taxa.The bark beetles exhibited a generally lower degree of spatial synchrony than the Lepidoptera, possibly because bark beetles are synchronized by different weather variables that are acting on a smaller scale than those affecting the Lepidoptera, or because inherent differences in their dynamics leads to more cyclic oscillations and hence more synchronous spatial dynamics in the Lepidoptera. Among the epidemic bark beetles tested, spatial synchrony of outbreaks in the Eurasian spruce bark beetle Ips typographus was significantly higher than for the other species.

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Abstract

The anatomical defense responses in stems of Norway spruce (Picea abies) clones of different resistance to pathogenic fungi were characterized over time and distance from small mechanical wounds or wounds inoculated with the root rot fungus Heterobasidion annosum. Common responses for both treatments included division of ray parenchyma and other cells in the cambial zone, accumulation of phenolic inclusions in ray parenchyma cells, activation of phloem parenchyma (PP) cells, and formation of traumatic resin ducts (TDs) in the xylem. TD formation occurred synchronously from a tangential layer of cells, or symplasmic domain, within the zone of xylem mother cells. TD induction is triggered by a signal, which propagates a developmental wave in the axial direction at about 2.5cm per day. TDs are formed at least 30cm above single inoculations within 16–36days after inoculation. The size and number of TDs is attenuated further away from the inoculation site, indicating a dose-dependent activity leading to TD development. Compared to sterile wounding, fungal inoculation gave rise to more and larger TDs in all clones, and multiple rows of TDs in weak clones. Fungal inoculation also induced the formation of more new PP cells, increasing the number of PP cells in the phloem in the year of inoculation up to 100%. TD and PP cell formation was greater in susceptible compared to resistant clones and after fungal versus sterile inoculation. Potential mechanisms responsible for this variable response are discussed.

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Abstract

Polyphenolic parenchyma cells (PP cells) in Norway spruce (Picea abies (L.) Karst.) stem phloem play important roles in constitutive and inducible defenses. To determine whether anatomical and molecular changes in PP cells are correlated with tree resistance, we infected two Norway spruce clones with the pathogenic fungus Ceratocystis polonica (Siem.) C. Moreau. The fungus induced significantly different lesion lengths in the two clones, indicating that one clone was more resistant to the fungus (short lesions) than the other (long lesions). After infection, the cross-sectional area of PP cells and their vacuolar polyphenol bodies increased in the three most recent annual rings of PP cells in both clones. The more resistant clone had larger PP cells with denser polyphenol bodies than the less resistant clone, whereas the less resistant clone accumulated relatively more polyphenols after infection. Compared with the less resistant clone, the more resistant clone contained higher starch concentrations before infection that were reduced more quickly after infection before returning to original values. Low transcript levels of chalcone synthase were detected in uninfected tissues of both clones, but the levels increased dramatically after infection. Transcript levels were higher and peaked 6 days earlier in the more resistant clone than in the less resistant clone. The activity of at least one highly basic peroxidase isoform was greatly enhanced after infection, and this increase occurred earlier in the more resistant clone.

Abstract

The study of conifer chemical defense has been dominated by investigations of oleoresin and its components. However, the actual function of resin components in plant defense and their mode of action is still uncertain, and the role of other defense compounds is relatively unexplored.We are studying the biochemical and molecular bases of chemical defenses, including terpenes, phenolics and chitinases, in Norway spruce (Picea abies) to learn more about how the accumulation of defense compounds is regulated, with the long-term goal of manipulating defense levels to test their function.Manipulation can be crudely accomplished by treatment with methyl jasmonate, which often mimics the general increases in defenses seen following herbivore or pathogen attack. Such treatment was shown to increase resistance to a fungal associate of bark beetles.To more conclusively test function, isolated genes of defense biosynthetic pathways are being transformed into Norway spruce to produce plants whose defense profiles are altered more precisely.

Abstract

When conifers such as Picea abies (Norway spruce) are attacked by insects or pathogens, they often produce increased quantities of terpenoid oleoresin. This response can be mimicked in young P. abies seedlings by treatment with methyl jasmonate. In this study, we determined the effect of methyl jasmonate on the terpenoids and other chemical defenses of mature P. abies, and investigated if this treatment protected trees against attack by the blue-stain fungus Ceratocystis polonica, the most important fungal associate of the bark beetle Ips typographus. Methyl jasmonate treatment induced the formation of traumatic resin ducts in the developing xylem, enhanced resin flow, and stimulated increased accumulation of monoterpenes, sesquiterpenes, and diterpene resin acids. However, almost no significant changes in terpene composition were detected. In addition, no changes in soluble phenolic content were observed. There was a very high variability both among and within clones in the timing and degree of response to methyl jasmonate. These chemical and anatomical changes were correlated with increased resistance to C. polonica, suggesting that terpenoid oleoresin may function in defense against this pathogen.

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Abstract

In 1996, 7000 ha of pine forests were defoliated by the pine looper Bupalus piniaria in south-western Sweden.The susceptibility of trees of different defoliation classes (0, 30, 60, 90 and 100% defoliation) to beetle-vectored blue-stain fungi was tested in inoculation experiments.Forty and 120-year-old Scots pine trees were inoculated with `single\", i.e. a few inoculations of Leptographium wingfieldii and Ophiostoma minus, two blue-stain fungi associated with the pine shoot beetle Tomicus piniperda. The young trees were also \"mass\" inoculated with L. wingfieldii at a density of 400 inoculation points per m2 over a 60 cm stem belt.Host tree symptoms indicated that only trees with 90100% defoliation were susceptible to the mass inoculation.Single inoculations did not result in any consistent differences in fungal performance between trees of different defoliation classes, regardless of inoculated species or tree age class.Leptographium wingfieldii produced larger reaction zones than O. minus, and both species produced larger lesions in old than in young trees.As beetle-induced tree mortality in the study area occurred only in totally defoliated stands, mass inoculations seem to mimic beetle-attacks fairly well, and thus seem to be a useful tool for assessing host resistance.As even severely defoliated pine trees were quite resistant, host defence reactions in Scots pine seem to be less dependent on carbon allocation than predicted by carbon-based defence hypotheses.

Abstract

The anatomical response of Norway spruce bark polyphenolic parenchyma cells (PP cells) to inoculation with the phytopathogenic fungus Ceratocystis polonica and attack by its bark-beetle vector Ips typographus was examined.Fungal inoculation on the periderm surface had no effect, while inoculation just below the periderm or halfway into the phloem (mid-phloem) generated detectable responses within 3 wk. The responses included increase in PP cell size and in periodic acid - Schiffs staining of PP cell phenolics, wound periderm initiation from PP cells, and cambial zone traumatic resin duct formation. Fungi were not seen in samples 3 wk after subperiderm or mid-phloem inoculation, but were found in some samples 6 and 9 wk after mid-phloem inoculation.In contrast, inoculations into the cambium resulted in partial (3 wk) or complete (6 and 9 wk) fungal colonization and death of tissue in the infected area. This indicates that PP cells have defenses capable of inhibiting fungal growth. Samples taken near bark-beetle galleries had similar anatomical responses as inoculated samples, validating the inoculation approach to studying defense responses in spruce.These results show that PP cells represent not only a constitutive defense system, but are also involved in local and remote inducible defenses against fungal and beetle attack.

Abstract

A field experiment was carried out to test the hypothesis that treatment of Norway spruce trees with the Ips typographus-transmitted blue-stain fungus Ceratocystis polonica enhances tree resistance to later mass attack by this bark beetle. Twenty-five mature trees were pre-treated by inoculating a non-lethal dose of the fungus into the bark, while 18 trees served as untreated controls. Three and a half weeks after treatment a bark beetle attack was initiated by attaching dispensers with I. typographus pheromone to the tree trunks. A significantly larger proportion (67%) of the control trees than of the pre-treated trees (36%) were killed by the beetle attack. The result is discussed in relation to recent results regarding defence mechanisms in Norway spruce trees.

Abstract

Twenty-five year old Norway spruce trees (Picea abies) were inoculated with four blue-stain fungi. Each tree was inoculated three times with each fungus and three times with sterile agar as a control, giving a total of 15 inoculations per tree. There was little variation in the extent of phloem necrosis produced in response to the different fungi, but five weeks after inoculation necrosis induced by Ceratocystis polonica and Ambrosiella sp. were significantly longer than those for the other fungi. At the same time, C. polonica had induced sapwood desicc-ation twice as deeply into the wood as any other fungus.Hyphal growth of the fungi into phloem and sapwood followed the same pattern as necrosis length and desiccation depth. Five weeks after inoculation C. polonica had penetrated phloem and sapwood further than any other fungus. It grew slower than the other fungi in both tissues the first week after inoculation, but the four following weeks it grew faster than all other fungi.

Abstract

Aggressive bark beetles kill healthy conifers through pheromone-mediated mass attacks. The exact mechanism by which trees are killed is still disputed, but phytopathogenic blue-stain fungi associated with the beetles are probably involved in most cases. This thesis compares the blue-stain flora of five bark beetle species that colonize Norway spruce of different resistance. The aggressive Ips typographus can kill healthy trees, while the other species are non-aggressive and colonize either severly stressed or dying trees (the facultatively parasitic Ips duplicatus, Polygraphus poligraphus and Pityogenes chalcographus), or dead trees (the saprophagic Hylurgops palliates). Ips typographus and I. duplicatus were both found to carry very high frequencies of the phytopathogenic fungus Ceratocystis polonica. This fungus has previously been found associated only with I. typographus, and has been shown to be pathogenic to Norway spruce in experimental mass inoculations. The other bark beetles carried no known pathogenic fungi. The phytopathogenicity of four of the isolated blue-stain fungi were evaluated through mass and low-density inoculation in young Norway spruce trees. Mass inoculations confirmed C. polonica\"s pathogenicity, while the other fungi, which are associated with non-aggressive beetles, were not pathogenic (Ophiostoma piceae, cfr. Ambrosiell sp., Dark sterile sp.A). Low-density inoculations revealed only small differences between the four fungi in phloem necrosis length. Necrosis length is used as a standard criterion of fungal pathogenicity, but it was not useful for assessing the pathogenicity of the fungi in the fungus-host tree system under study here. Ceratocystis polonica was found to penertrate deeper into the sapwood and induce deeper sapwood desiccation than the other fungi. The ability to invade sapwood is probably more important for fungal pathogenicity than the ability to colonize phloem, and may thus be a better criterion for assessing the pathogenicity of blue-stain fungi. General aspects of the association between bark beetles and blue-stain fungi are discussed.