Defense of Scots pine against sawfly eggs (Diprion pini) is primed by exposure to sawfly sex pheromones.

Plants respond to insect infestation with defenses targeting insect eggs on their leaves and the feeding insects. Upon perceiving cues indicating imminent herbivory, such as damage-induced leaf odors emitted by neighboring plants, they are able to prime their defenses against feeding insects. Yet it remains unknown whether plants can amplify their defenses against insect eggs by responding to cues indicating imminent egg deposition. Here, we tested the hypothesis that a plant strengthens its defenses against insect eggs by responding to insect sex pheromones. Our study shows that preexposure of Pinus sylvestris to pine sawfly sex pheromones reduces the survival rate of subsequently laid sawfly eggs. Exposure to pheromones does not significantly affect the pine needle water content, but results in increased needle hydrogen peroxide concentrations and increased expression of defense-related pine genes such as SOD (superoxide dismutase), LOX (lipoxygenase), PAL (phenylalanine ammonia lyase), and PR-1 (pathogenesis related protein 1) after egg deposition. These results support our hypothesis that plant responses to sex pheromones emitted by an herbivorous insect can boost plant defensive responses to insect egg deposition, thus highlighting the ability of a plant to mobilize its defenses very early against an initial phase of insect attack, the egg deposition.

Identifying the abiotic and biotic drivers behind the elevational distribution shift of a parasitic plant.

Climate change will alter the biotic and abiotic environment and dissipate ecological barriers, reorganising maps of current distribution of parasites and their hosts. In this study, we analyse the population dynamics of the parasitic plant Viscum album subsp. austriacum and explore key biotic (host availability and seed dispersal) as well as abiotic (temperature) factors influencing elevational distribution. The study was conducted along an elevational gradient of a Mediterranean mountain, covering the distribution belts of three potential pine hosts: Pinus halepensis (1300-1500 m), P. nigra (1300-1900 m) and P. sylvestris var. nevadensis (1600-2000 m). Along this gradient, we measured multiple variables of mistletoe population (prevalence, abundance and demographic profile) and different factors that might define the current mistletoe distribution (host suitability and availability, temperature and seed dispersal services). We found a decline in mistletoe prevalence and abundance with increasing elevation, detecting larger values of both variables at lower elevations of the most suitable host (Pinus nigra). Pinus sylvestris var. nevadensis was a suboptimal but suitable host for the parasite at high elevations. Mistletoe found suitable temperatures and seed dispersal services all along the gradient, being able to recruit at any site. With warming temperatures, the presence of suitable vectors for parasite dispersion, and the presence of a sub-optimal host (P. sylvestris var. nevadensis) at the mountain top, mistletoe currently has a window of opportunity to expand its present geographic distribution to the summits.

We Are What We Eat: A Stoichiometric and Ecometabolomic Study of Caterpillars Feeding on Two Pine Subspecies of Pinus sylvestris.

Many studies have addressed several plant-insect interaction topics at nutritional, molecular, physiological, and evolutionary levels. However, it is still unknown how flexible the metabolism and the nutritional content of specialist insect herbivores feeding on different closely related plants can be. We performed elemental, stoichiometric, and metabolomics analyses on leaves of two coexisting Pinus sylvestris subspecies and on their main insect herbivore; the caterpillar of the processionary moth (Thaumetopoea pityocampa). Caterpillars feeding on different pine subspecies had distinct overall metabolome structure, accounting for over 10% of the total variability. Although plants and insects have very divergent metabolomes, caterpillars showed certain resemblance to their plant-host metabolome. In addition, few plant-related secondary metabolites were found accumulated in caterpillar tissues which could potentially be used for self-defense. Caterpillars feeding on N and P richer needles had lower N and P tissue concentration and higher C:N and C:P ratios, suggesting that nutrient transfer is not necessarily linear through trophic levels and other plant-metabolic factors could be interfering. This exploratory study showed that little chemical differences between plant food sources can impact the overall metabolome of specialist insect herbivores. Significant nutritional shifts in herbivore tissues could lead to larger changes of the trophic web structure.

Insectivorous Birds Are Attracted by Plant Traits Induced by Insect Egg Deposition.

Insectivorous birds feed upon all developmental stages of herbivorous insects, including insect eggs if larvae and adults are unavailable. Insect egg deposition on plants can induce plant traits that are subsequently exploited by egg parasitoids searching for hosts. However, it is unknown whether avian predators can also use egg-induced plant changes for prey localization. Here, we studied whether great tits (Parus major) and blue tits (Cyanistes caeruleus) are attracted by traits of the Scots pine (Pinus sylvestris) induced by pine sawfly (Diprion pini) egg deposition. We chose this plant - insect system because sawfly egg deposition on pine needles is known to locally and systemically induce a change in pine volatile organic compounds (VOCs), and tits are known to prey upon sawfly eggs. In dual choice laboratory experiments, we simultaneously offered the birds an egg-free control branch and a systemically egg-induced branch. Significantly more birds visited the egg-induced branch first. We confirmed by GC-MS analyses that systemically egg-induced branches released more (E)-ß-farnesene compared to control branches. Spectrophotometric analyses showed that control branches reflected more light than egg-induced branches throughout the avian visual range. Although a discrimination threshold model for blue tits suggests that the birds are poor at discriminating this visual difference, the role of visual stimuli in attracting the birds to egg-induced pines cannot be discounted. Our study shows, for the first time, that egg-induced odorous and/or visual plant traits can help birds to locate insect eggs without smelling or seeing those eggs.

Mistletoe-induced growth reductions at the forest stand scale.

The hemiparasite European mistletoe (Viscum album L.) adversely affects growth and reproduction of the host Scots pine (Pinus sylvestris L.) and in consequence may lead to tree death. Here, we aimed to estimate mistletoe-induced losses in timber yield applying the process-based forest growth model 4C. The parasite was implemented into the eco-physiological forest growth model 4C using (literature-derived) established impacts of the parasite on the tree's water and carbon cycle. The amended model was validated simulating a sample forest stand in the Berlin area (Germany) comprising trees with and without mistletoe infection. At the same forest stand, tree core measurements were taken to evaluate simulated and observed growth. A subsample of trees were harvested to quantify biomass compartments of the tree canopy and to derive a growth function of the mistletoe population. The process-based simulations of the forest stand revealed 27% reduction in basal area increment (BAI) during the last 9 years of heavy infection, which was confirmed by the measurements (29% mean growth reduction). The long-term simulations of the forest stand before and during the parasite infection showed that the amended forest growth model 4C depicts well the BAI growth pattern during >100 years and also quantifies well the mistletoe-induced growth reductions in Scots pine stands.

Mistletoe infestation mediates alteration of the phytohormone profile and anti-oxidative metabolism in bark and wood of its host Pinus sylvestris.

European mistletoe (Viscum album L.) has largely infested Central European forests and causes high mortality probability particularly in dry years. However, little information is available about the consequences of mistletoe infestation for metabolic processes in bark and wood of its host, despite their important roles in infestation defense. We analyzed the tissue hydration, carbohydrate composition, phytohormone profile, reactive oxygen species and anti-oxidant levels in bark and wood of Scots pines (Pinus sylvestris L.), as dependent on mistletoe infestation. As a consequence of mistletoe infestation, host bark and wood showed impaired hydration and reduced total carbon content. In the bark, soluble sugar and lignin contents increased, apparently at the expense of holo-cellulose. Hydrogen peroxide accumulation was accompanied by increased glutathione and decreased reduced ascorbic acid levels. Mistletoe infestation mediated alteration of the phytohormone profile in bark and wood of its host. Cytokinins, jasmonic acid and abscisic acid levels increased in both tissues, whereas salicylic acid and indole-3-acetic acid, which were only detected in the bark, declined. The present results show that mistletoe infestation affects both the host's anti-oxidative defense system and the phytohormone profile after establishment of the xylem tapping haustorium. The significance of these processes for the development of the woody mistletoe stem and the haustorium is discussed.

Early plant defence against insect attack: involvement of reactive oxygen species in plant responses to insect egg deposition.

MAIN CONCLUSION: Pinus sylvestris responds to insect egg deposition by ROS accumulation linked with reduced activity of the ROS scavenger catalase. Egg mortality in needles with hypersensitive response (HR)-like symptoms is enhanced. Aggressive reactive oxygen species (ROS) play an important role in plant defence against biotic stressors, including herbivorous insects. Plants may even generate ROS in response to insect eggs, thus effectively fighting against future larval herbivory. However, so far nothing is known on how ROS-mediated plant defence against insect eggs is enzymatically regulated. Neither do we know how insects cope with egg-induced plant ROS. We addressed these gaps of knowledge by studying the activities of ROS-related enzymes in Pinus sylvestris deposited with eggs of the herbivorous sawfly Diprion pini. This species cuts a slit into pine needles and inserts its eggs into the needle tissue. About a quarter of egg-deposited needles show chlorotic tissue at the oviposition sites, indicating hypersensitive response-like direct defence responses resulting in reduced larval hatching from eggs. Hydrogen peroxide and peroxidase sensitive staining of sections of egg-deposited pine needles revealed the presence of hydrogen peroxide and peroxidase activity in needle tissue close to the eggs. Activity of ROS-producing NADPH-oxidase did not increase after egg deposition. However, the activity of the ROS-detoxifying enzyme catalase decreased after egg deposition and ovipositional wounding of needles. These results show that local ROS accumulation at the oviposition site is not caused by increased NADPH-oxidase activity, but reduced activity of pine needle catalase may contribute to it. However, our data suggest that pine sawflies can counteract the egg deposition-induced hydrogen peroxide accumulation in pine needles by high catalase activity in their oviduct secretion which is released with the eggs into pine tissue.

Form resemblance between the white-berried mistletoe (Viscum album L. ) and five host trees / Correspondências morfológicas entre o Viscum album L. e cinco árvores hospedeiras

The development of new forms of thinking, that enables us to participate in the creative processes of nature, became one of the most important tasks for the present time and the future. With this aim, the author presents his research in the Natural Science Section at the Goetheanum (Glashaus). On Goethian basis, the correspondence between white-berried mistletoe (Viscum album L.) and its host tree (Malus domestica, Ulmus minor, Quercus robur, Tilia cordata and Pinus silvestris) was investigated, searching what kind of host tree qualities are expressed by the different mistletoe forms. The methodology was: field observation, photography, artistic exercises and literature research. Some general aspects of the five trees species and the respective mistletoe types were portrayed in terms of their Gestalt. Finally it was compared the types, looking for the polarities that can serve as a starting point for an understanding of the qualitative relationship between mistletoe and host tree. It was possible to gather some evidences to support the two hypotheses: there are resemblances in terms of Gestalt between host trees and mistletoes and there are polarities between the mistletoe varieties.(AU)

O desenvolvimento de novas formas de pensar, que nos permite participar dos processos criativos da natureza, tornou-se uma das tarefas mais importantes para nossa época e para o futuro. Com este objetivo, o autor apresenta sua pesquisa na Seção de Ciências Naturais no Goetheanum (Glashaus). Em bases goethianas, foi investigada a correspondência entre o Viscum album L. e suas árvores hospedeiras (Malus domestica, Ulmus minor, Quercus robur, Tilia cordata e Pinus silvestris), buscando quais tipos de qualidades da árvore hospedeira são expressas pelas diferentes formas de Viscum album. A metodologia foi: observação de campo, fotografia, exercícios artísticos e pesquisa literária. Alguns aspectos gerais das cinco espécies de árvores hospedeiras e os respectivos tipos de Viscum foram retratados em termos de 'Gestalt'. Finalmente, foram comparados os tipos, buscando polaridades que podem servir de ponto de partida para a compreensão da relação qualitativa entre o Viscum e a árvore hospedeira. Foi possível reunir algumas evidências para apoiar as duas hipóteses: há semelhanças em termos de 'Gestalt' entre as árvores hospedeiras e o Viscum album e há polaridades entre as variedades de Viscum album.(AU)

Methyl Jasmonate-Induced Monoterpenes in Scots Pine and Norway Spruce Tissues Affect Pine Weevil Orientation.

In large parts of Europe, insecticide-free measures for protecting conifer plants are desired to suppress damage by the pine weevil Hylobius abietis (L.). Treatment with methyl jasmonate (MeJA), a chemical elicitor already used in crop production, may enhance expression of chemical defenses in seedlings in conifer regenerations. However, in a previous experiment, MeJA treatment resulted in substantially better field protection for Scots pine (Pinus sylvestris L.) than for Norway spruce (Picea abies (L.) Karst.). Hypothesizing that the variations may be at least due partly to volatiles released by MeJA-treated seedlings and their effects on pine weevil orientation, we examined tissue extracts of seedlings (from the same batches as previously used) by two-dimensional GC-MS. We found that the MeJA treatment increased contents of the monoterpene (-)-ß-pinene in phloem (the weevil's main target tissue) of both tree species, however, the (-)-ß-pinene/(-)-α-pinene ratio increased more in the phloem of P. sylvestris. We also tested the attractiveness of individual monoterpenes found in conifer tissues (needles and phloem) for pine weevils using an arena with traps baited with single-substance dispensers and pine twigs. Trap catches were reduced when the pine material was combined with a dispenser releasing (-)-ß-pinene, (+)-3-carene, (-)-bornyl acetate or 1,8-cineole. However, (-)-α-pinene did not have this effect. Thus, the greater field protection of MeJA-treated P. sylvestris seedlings may be due to the selective induction of increases in contents of the deterrent (-)-ß-pinene, in contrast to strong increases in both non-deterrent (-)-α-pinene and the deterrent (-)-ß-pinene in P. abies seedlings.

Effects of mistletoe removal on growth, N and C reserves, and carbon and oxygen isotope composition in Scots pine hosts.

Most mistletoes are xylem-tapping hemiparasites, which derive their resources from the host's xylem solution. Thus, they affect the host's water relations and resource balance. To understand the physiological mechanisms underlying the mistletoe-host relationship, we experimentally removed Viscum album ssp. austriacum (Wiesb.) Vollmann from adult Pinus sylvestris L. host trees growing in a Swiss dry valley. We analyzed the effects of mistletoe removal over time on host tree growth and on concentrations of nonstructural carbohydrates (NSC) and nitrogen (N) in needles, fine roots and sapwood. In addition, we assessed the δ(13)C and δ(18)O in host tree rings. After mistletoe removal, δ(13)C did not change in newly produced tree rings compared with tree rings in control trees (still infected with mistletoe), but δ(18)O values increased. This pattern might be interpreted as a decrease in assimilation (A) and stomatal conductance (gs), but in our study, it most likely points to an inadequacy of the dual isotope approach. Instead, we interpret the unchanged δ(13)C in tree rings upon mistletoe removal as a balanced increase in A and gs that resulted in a constant intrinsic water use efficiency (defined as A/gs). Needle area-based concentrations of N, soluble sugars and NSC, as well as needle length, single needle area, tree ring width and shoot growth, were significantly higher in trees from which mistletoe was removed than in control trees. This finding suggests that mistletoe removal results in increased N availability and carbon gain, which in turn leads to increased growth rates of the hosts. Hence, in areas where mistletoe is common and the population is large, mistletoe management (e.g., removal) may be needed to improve the host vigor, growth rate and productivity, especially for relatively small trees and crop trees in xeric growth conditions.

Mistletoe (Viscum album) infestation in the Scots pine stimulates drought-dependent oxidative damage in summer.

This study sought to contribute to the understanding of the detrimental effect of the mistletoe (Viscum albumL.), a hemiparasitic plant, on the mortality of the Scots pine (Pinus sylvestrisL.). Fieldwork was conducted in the town of Kelkit (Gumushane province, Turkey) from April to October in 2013. Pine needles of similar ages were removed from the branches of mistletoe-infested and noninfested Scots pine plants, then transported to the laboratory and used as research materials. The effects of the mistletoe on the Scots pine during infestation were evaluated by determining the levels of water, electrolyte leakage (EL), malondialdehyde (MDA, being a product of lipid peroxidation) and reactive oxygen species (ROS) such as superoxide anion (O2 (-•)), hydrogen peroxide (H2O2) and hydroxyl radical ((•)OH). In addition, the activities of antioxidative enzymes such as superoxide dismutase (SOD), catalase (CAT) and peroxidase (POX) were measured in the same samples. The highest level of drought stress was found in summer (especially in August) as a result of the lowest water content in the soil and the highest average temperature occurring in these months. The drought stress induced by mistletoe infestation caused a regular decrease in water content, while it increased the levels of EL, MDA and ROS (H2O2, O2 (-•)and(•)OH). The infestation also stimulated the activities of CAT and POX, with the exception of SOD. On the other hand, in August, when the drought conditions were the harshest, the levels of EL and MDA, which are two of the most important indicator parameters for oxidative stress, as well as the levels of H2O2and(•)OH, which are two of the ROS leading to oxidative stress, reached the highest values in both infested and noninfested needles, whereas the O2 (-•)level decreased. For the same period and needles, CAT activity increased, while SOD activity decreased. Peroxidase activity, however, did not exhibit a significant change. Our findings indicate that the increased mortality of the Scots pine may result from the mistletoe-induced very severe drought stress, and that the increase in the capacity of antioxidative enzyme system does not protect the plant against oxidative stress in dry summer seasons.

Nutritional and pathogenic fungi associated with the pine engraver beetle trigger comparable defenses in Scots pine.

Conifer bark beetles are often associated with fungal complexes whose components have different ecological roles. Some associated species are nutritionally obligate fungi, serving as nourishment to the larvae, whereas others are pathogenic blue-stain fungi known to be involved in the interaction with host defenses. In this study we characterized the local and systemic defense responses of Scots pine (Pinus sylvestris L.) against Ophiostoma brunneo-ciliatum Math. (a blue-stain pathogen) and Hyalorhinocladiella macrospora (Franke-Grosm.) Harr. (a nutritional fungus). These fungi are the principal associates of the pine engraver beetle, Ips acuminatus (Gyll.). Host responses were studied following inoculation with the fungi, singly and as a fungal complex, and by identifying and quantifying terpenoids, phenolic compounds and lignin. Although the length of the necrotic lesions differed between control (wound) and fungal treatments, only two compounds (pinosylvin monomethyl ether and (+)-α-pinene) were significantly affected by the presence of the fungi, indicating that Scots pine has a generic, rather than specific, induced response. The fact that both nutritional and blue-stain fungi triggered comparable induced defense responses suggests that even a non-pathogenic fungus may participate in exhausting host plant defenses, indirectly assisting in the beetle establishment process. Our findings contribute to the further development of current theory on the role of associated fungal complexes in bark beetle ecology.

Putative origin of clonal lineages of Amylostereum areolatum, the fungal symbiont associated with Sirex noctilio, retrieved from Pinus sylvestris, in eastern Canada.

The Eurasian Sirex noctilio-Amylostereum areolatum complex was discovered and has become established close to the North American Great Lakes in the 2000s. This invasive forest insect pest represents a very high risk to native and exotic pines in North America. We investigated the geographical origin of clonal lineages of the fungal symbiont A. areolatum in the recently pest-colonized eastern Canadian region by analyzing mitochondrial and nuclear sequence variations and comparing the genetic diversity of a worldwide collection of fungal symbionts among six countries where the Sirex complex is native and four countries from which the insect-fungal complex has been introduced. In total, 102 isolates were analyzed. While 12 multilocus genotypes (MLGs) are observed in the areas where S. noctilio is native, only two MLGs are retrieved from areas where S. noctilio is not native, indicating the wide spread of clonal lineages in the introduced fungal symbiont of S. noctilio. MLG2 comprises 26% of the Canadian isolates and is also observed in Chile and South Africa, where the insect-fungal complex has also been introduced. MLG3 comprises 74% of the Canadian isolates and is also observed in the USA, but nowhere else in our worldwide collection. Thus, at least one of the Canadian clonal lineages shares a common origin with A. areolatum isolates from the Southern Hemisphere. The source of the second clonal lineage is still unknown, but phylogenetic analyses show that MLG3 is isolated. More extended sampling is necessary to determine the origin of this fungal clonal lineage and investigate its probable symbiotic association with native North American Sirex.

Within-tree distributions of the Sirex noctilio Fabricius (Hymenoptera: Siricidae)--parasitoid complex and development of an optimal sampling scheme.

Little is known about the introduced European woodwasp, Sirex noctilio F. (Hymenoptera: Siricidae), and its hymenopteran parasitoid complex in North America. To assess within-tree and landscape-level densities and distributions of these insects, and develop a more cost effective approach for sampling and monitoring S. noctilio, 18 infested pines (12 P. resinosa Aiton and six P. sylvestris L.) were felled from six stands within three regions in central New York and cut into 0.5-m bolts. Bolts were carefully split to recover all siricids (S. noctilio and native siricids) and parasitoids. In total, 2,558 siricids were recovered; 1,972 siricid larvae and 586 teneral adult S. noctilio. Parasitism of siricids, a majority of which were S. noctilio based on larval rearing results, was 16.4% with Ibalia leucospoides ensiger Norton causing 10.7% of the documented mortality. Numbers of siricids and parasitoids declined 33-86% from the northern to the southern sampling regions, peak insect densities occurred in sections of the bole 15-19 cm in diameter and numbers of insects were generally higher in P. sylvestris than P. resinosa according to the highest ranked zero-inflated poisson and probit regression models. Bark thickness was not correlated with siricid or parasitoid species densities. We also describe sampling plans where as few as two 0.5-m samples from infested trees provided reliable within-tree insect densities.

Mistletoe-induced crown degradation in Scots pine in a xeric environment.

Increasing Scots pine (Pinus sylvestris L.) mortality has been recently observed in the dry inner valleys of the European Alps. Besides drought, infection with pine mistletoe (Viscum album ssp. austriacum) seems to play an important role in the mortality dynamics of Scots pines, but how mistletoes promote pine decline remains unclear. To verify whether pine mistletoe infection weakens the host via crown degradation, as observed for dwarf mistletoes, we studied the negative effects of pine mistletoe infestation on the photosynthetic tissues and branch growth of pairs of infested and non-infested branches. Pine mistletoe infection leads to crown degradation in its host by reducing the length, the radial increment, the ramification, the needle length and the number of needle years of the infested branches. This massive loss in photosynthetic tissue results in a reduction in primary production and a subsequent decrease in carbohydrate availability. The significant reduction in needle length due to mistletoe infection is an indication for a lower water and nutrient availability in infested branches. Thus, mistletoe infection might lead to a decrease in the availability of water and carbohydrates, the two most important growth factors, which are already shortened due to the chronic drought situation in the area. Therefore, pine mistletoe increases the risk of drought-induced mortality of its host when growing in a xeric environment.

Reduction of ethylene emission from Scots pine elicited by insect egg secretion.

Pinus sylvestris L. is known to activate indirect defence in response to attack by an herbivorous sawfly. Egg deposition by the sawfly Diprion pini L. induces pine to release, three days after egg laying, locally and systemically terpenoid volatiles that attract parasitoids to kill the eggs. The elicitor of the pine's response is located in the sawfly's oviduct secretion enveloping the eggs after deposition. Application of this secretion on twigs with artificially conducted ovipositional woundings mimics the effects of egg deposition. Furthermore, jasmonic acid (JA) induces a volatile pattern similar, but not identical, to the one induced by egg deposition. To gain deeper insight into the transduction of plant signals induced by herbivore egg deposition, it was investigated whether ethylene emission from pine is affected by sawfly egg deposition. Systemically induced ethylene emission from differently treated pine twigs was monitored for a period of 3 d after treatment. Ethylene emissions from untreated control twigs were compared with those from twigs treated as follows: (i) sawfly egg secretion [=oviduct secretion (OVI)] was transferred on artificially wounded pine needles (attractive volatiles), (ii) needles were artificially wounded (non-attractive volatiles), and (iii) the twig was supplied with JA (attractive volatiles). Ethylene emission from systemically OVI-induced twigs was significantly lower than from untreated controls, whereas artificial wounding had no detectable effect. JA-treated twigs released much more ethylene and showed higher variability of ethylene emission than artificially wounded twigs and OVI-treated ones. Ethylene emissions from pine after the various treatments studied here are discussed with respect to known effects of insect feeding on ethylene release from plants.

Linking increasing drought stress to Scots pine mortality and bark beetle infestations.

In the dry Swiss Rhone Valley, Scots pine forests have experienced increased mortality in recent years. It has commonly been assumed that drought events and bark beetles fostered the decline, however, whether bark beetle outbreaks increased in recent years and whether they can be linked to drought stress or increasing temperature has never been studied. In our study, we correlated time series of drought indices from long-term climate stations, 11-year mortality trends from a long-term research plot, and mortality probabilities modeled from tree rings (as an indicator of tree vitality) with documented occurrences of various bark beetle species and a buprestid beetle, using regional Forest Service reports from 1902 to 2003 and advisory cases of the Swiss Forest Protection Service (SFPS) from 1984 to 2005. We compared the historical findings with measured beetle emergence from a 4-year tree felling and breeding chamber experiment. The documented beetle-related pine mortality cases increased dramatically in the 1990s, both in the forest reports and the advisory cases. The incidents of beetle-related pine mortality correlated positively with spring and summer temperature, and with the tree-ring based mortality index, but not with the drought index. The number of advisory cases, on the other hand, correlated slightly with summer drought index and temperature, but very highly with tree-ring-based mortality index. The tree-ring-based mortality index and observed tree mortality increased in years following drought. This was confirmed by the beetle emergences from felled trees. Following dry summers, more than twice as many trees were colonized by beetles than following wet summers. We conclude that increased temperatures in the Swiss Rhone Valley have likely weakened Scots pines and favored phloeophagous beetle population growth. Beetles contributed to the increased pine mortality following summer drought. Among the factors not addressed in this study, changed forest use may have also contributed to increased beetle populations and Scots pine mortality, whereas air pollution seems to be of lesser importance.

The role of genetic and chemical variation of Pinus sylvestris seedlings in influencing slug herbivory.

This study investigated the genetic and chemical basis of resistance of Pinus sylvestris seedlings to herbivory by a generalist mollusc, Arion ater. Using feeding trials with captive animals, we examined selective herbivory by A. ater of young P. sylvestris seedlings of different genotypes and correlated preferences with seedling monoterpene levels. We also investigated the feeding responses of A. ater to artificial diets laced with two monoterpenes, Delta(3)-carene and alpha-pinene. Logistic regression indicated that two factors were the best predictors of whether seedlings in the trial would be consumed. Individual slug variation (replicates) was the most significant factor in the model; however, alpha-pinene concentration (also representing beta-pinene, Delta(3)-carene and total monoterpenes due to multicollinearity) of needles was also a significant factor. While A. ater did not select seedlings on the basis of family, seedlings not eaten were significantly higher in levels of alpha-pinene compared to seedlings that were consumed. We also demonstrated significant genetic variation in alpha-pinene concentration of seedlings between different families of P. sylvestris. Nitrogen and three morphological seedling characteristics (stem length, needle length and stem diameter) also showed significant genetic variation between P. sylvestris families. Artificial diets laced with high (5 mg g(-1) dry matter) quantities of either Delta(3)-carene or alpha-pinene, were eaten significantly less than control diets with no added monoterpenes, supporting the results of the seedling feeding trial. This study demonstrates that A. ater selectively feed on P. sylvestris seedlings and that this selection is based, in part, on the monoterpene concentration of seedlings. These results, coupled with significant genetic variation in alpha-pinene concentration of seedlings and evidence that slug herbivory is detrimental to P. sylvestris fitness, are discussed as possible evidence for A. ater as a selective force on the evolution of defensive chemistry in P. sylvestris.

Genetic differentiation in Scottish populations of the pine beauty moth, Panolis flammea (Lepidoptera: Noctuidae).

Pine beauty moth, Panolis flammea (Denis & Schiffermüller), is a recent but persistent pest of lodgepole pine plantations in Scotland, but exists naturally at low levels within remnants and plantations of Scots pine. To test whether separate host races occur in lodgepole and Scots pine stands and to examine colonization dynamics, allozyme, randomly amplified polymorphic DNA (RAPD) and mitochondrial variation were screened within a range of Scottish samples. RAPD analysis indicated limited long distance dispersal (FST=0.099), and significant isolation by distance (P<0.05); but that colonization between more proximate populations was often variable, from extensive to limited exchange. When compared with material from Germany, Scottish samples were found to be more diverse and significantly differentiated for all markers. For mtDNA, two highly divergent groups of haplotypes were evident, one group contained both German and Scottish samples and the other was predominantly Scottish. No genetic differentiation was evident between P. flammea populations sampled from different hosts, and no diversity bottleneck was observed in the lodgepole group. Indeed, lodgepole stands appear to have been colonized on multiple occasions from Scots pine sources and neighbouring populations on different hosts are close to panmixia.

The significance of background odour for an egg parasitoid to detect plants with host eggs.

Scots pine has been shown to produce a volatile bouquet that attracts egg parasitoids in response to oviposition of the herbivorous sawfly Diprion pini. Previous analyses of headspace volatiles of oviposition-induced pine twigs revealed only quantitative changes; in particular, the sesquiterpene (E)-beta-farnesene was emitted in significantly higher quantities by oviposition-induced pine. Here we investigated whether (E)-beta-farnesene attracted the egg parasitoid Chrysonotomyia ruforum. We tested the behavioural response of C. ruforum females to different concentrations of (E)-beta-farnesene. Egg parasitoids did not respond to this sesquiterpene at either concentration tested. However, they did respond significantly to (E)-beta-farnesene when this compound was offered in combination with the volatile blend emitted from pine twigs without eggs. This response was dependent on the applied concentration of (E)-beta-farnesene. Further bioassays with other components [(E)-beta-caryophyllene, delta-cadinene] of the odour blend of pine were conducted in combination with the volatile blend from egg-free pine as background odour. None of the compounds tested against the background of odour from an egg-free pine twig were attractive to the egg parasitoid. These results suggest that the egg parasitoids responded specifically to (E)-beta-farnesene, but only when this compound was experienced in the 'right' context, i.e. when contrasted with a background odour of non-oviposition-induced pine volatiles.