Primary and secondary host selection by Ips typographus depends on Norway spruce crown characteristics and phenolic-based defenses.

Climate change is expected to intensify bark beetle population outbreaks in forests globally, affecting biodiversity and trajectories of change. Aspects of individual tree resistance remain poorly quantified, particularly with regard to the role of phenolic compounds, hindering robust predictions of forest response to future conditions. In 2003, we conducted a mechanical wounding experiment in a Norway spruce forest that coincided with an outbreak of the bark beetle, Ips typographus. We collected phloem samples from 97 trees and monitored tree survival for 5 months. Using high-performance liquid chromatography, we quantified induced changes in the concentrations of phenolics. Classification and regression tools were used to evaluate relationships between phenolic production and bark beetle resistance, in the context of other survival factors. The proximity of beetle source populations was a principal determinant of survival. Proxy measures of tree vigor, such as crown defoliation, mediated tree resistance. Controlling for these factors, synthesis of catechin was found to exponentially increase tree survival probability. However, even resistant trees were susceptible in late season due to high insect population growth. Our results show that incorporating trait-mediated effects improves predictions of survival. Using an integrated analytical approach, we demonstrate that phenolics play a direct role in tree defense to herbivory.

Sentinel nurseries to assess the phytosanitary risks from insect pests on importations of live plants.

Ornamental plants represent a major pathway of invasion for alien pests worldwide. Commodity risk analyses are carried out to assess the risk posed by a new trade in a commodity, but they are restricted by our limited knowledge of the pests carried by traded plants. We used the sentinel nursery technique to identify insects attacking woody plants imported into Europe. We established two sentinel nurseries in China, with five traded Asian plants. These nurseries were monitored for two years to obtain lists of insects that can be expected on these commodities. These records were compared with those obtained from literature surveys, which are usually the sources of information available to pest risk assessors. At each site, 105 insect species and host associations were found on sentinel plants and 90% of these associations had not been found in a previous literature survey of insect pests of the five plants. Nearly 80% of these associations were not found in an a posteriori literature survey. An assessment classified 9%, 7% and 84% of the insect records as presenting a high, moderate and low likelihood of introduction, respectively. These results show the benefit of sentinel nurseries to identify potential infestation of plant commodity imports.

Planting sentinel European trees in eastern Asia as a novel method to identify potential insect pest invaders.

Quarantine measures to prevent insect invasions tend to focus on well-known pests but a large proportion of the recent invaders were not known to cause significant damage in their native range, or were not even known to science before their introduction. A novel method is proposed to detect new potential pests of woody plants in their region of origin before they are introduced to a new continent. Since Asia is currently considered to be the main supplier of insect invaders to Europe, sentinel trees were planted in China during 2007-2011 as an early warning tool to identify the potential for additional Asian insect species to colonize European trees. Seedlings (1-1.5 m tall) of five broadleaved (Quercus petraea, Q. suber, Q. ilex, Fagus sylvatica, and Carpinus betulus) and two conifer species (Abies alba and Cupressus sempervirens) were planted in blocks of 100 seedlings at two widely separated sites (one in a nursery near Beijing and the other in a forest environment near Fuyang in eastern China), and then regularly surveyed for colonization by insects. A total of 104 insect species, mostly defoliators, were observed on these new hosts, and at least six species were capable of larval development. Although a number of the insects observed were probably incidental feeders, 38 species had more than five colonization events, mostly infesting Q. petraea, and could be considered as being capable of switching to European trees if introduced to Europe. Three years was shown to be an appropriate duration for the experiment, since the rate of colonization then tended to plateau. A majority of the identified species appeared to have switched from agricultural crops and fruit trees rather than from forest trees. Although these results are promising, the method is not appropriate for xylophagous pests and other groups developing on larger trees. Apart from the logistical problems, the identification to species level of the specimens collected was a major difficulty. This situation could be improved by the development of molecular databases.

Sentinel trees as a tool to forecast invasions of alien plant pathogens.

Recent disease outbreaks caused by alien invasive pathogens into European forests posed a serious threat to forest sustainability with relevant environmental and economic effects. Many of the alien tree pathogens recently introduced into Europe were not previously included on any quarantine lists, thus they were not subject to phytosanitary inspections. The identification and description of alien fungi potentially pathogenic to native European flora before their introduction in Europe, is a paramount need in order to limit the risk of invasion and the impact to forest ecosystems. To determine the potential invasive fungi, a sentinel trees plot was established in Fuyang, China, using healthy seedlings of European tree species including Quercus petreae, Q. suber, and Q. ilex. The fungal assemblage associated with symptomatic specimens was studied using the tag-encoded 454 pyrosequencing of the nuclear ribosomal internal transcribed spacer-1 (ITS 1). Taxa with probable Asiatic origin were identified and included plant pathogenic genera. These results indicate that sentinel plants may be a strategic tool to improve the prevention of bioinvasions.

A suite of models to support the quantitative assessment of spread in pest risk analysis.

Pest Risk Analyses (PRAs) are conducted worldwide to decide whether and how exotic plant pests should be regulated to prevent invasion. There is an increasing demand for science-based risk mapping in PRA. Spread plays a key role in determining the potential distribution of pests, but there is no suitable spread modelling tool available for pest risk analysts. Existing models are species specific, biologically and technically complex, and data hungry. Here we present a set of four simple and generic spread models that can be parameterised with limited data. Simulations with these models generate maps of the potential expansion of an invasive species at continental scale. The models have one to three biological parameters. They differ in whether they treat spatial processes implicitly or explicitly, and in whether they consider pest density or pest presence/absence only. The four models represent four complementary perspectives on the process of invasion and, because they have different initial conditions, they can be considered as alternative scenarios. All models take into account habitat distribution and climate. We present an application of each of the four models to the western corn rootworm, Diabrotica virgifera virgifera, using historic data on its spread in Europe. Further tests as proof of concept were conducted with a broad range of taxa (insects, nematodes, plants, and plant pathogens). Pest risk analysts, the intended model users, found the model outputs to be generally credible and useful. The estimation of parameters from data requires insights into population dynamics theory, and this requires guidance. If used appropriately, these generic spread models provide a transparent and objective tool for evaluating the potential spread of pests in PRAs. Further work is needed to validate models, build familiarity in the user community and create a database of species parameters to help realize their potential in PRA practice.

Setae from the pine processionary moth (Thaumetopoea pityocampa) contain several relevant allergens.

BACKGROUND: Pine processionary larvae produce urticating hairs (setae) that serve for protection against predators. Setae induce cutaneous reactions in animals and humans. The presence of toxic or allergic mechanisms is a matter of debate. OBJECTIVES: To detect the presence of allergens in setae and to characterize them. MATERIALS AND METHODS: Setae extracts were characterized by gel staining and immunoblot, with sera from patients with immediate reactions and positive prick test reactions, as well as a rabbit antiserum raised against setae. Setae proteins were fractionated by high-performance liquid chromatography. The most relevant allergen was analysed by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS), and its sequence was deduced from an expressed sequence tag bank. Results. Setae contained at least seven different allergens. The most intense detection corresponded to a protein of MW ~ 14,000 that was similar to thaumetopoein, a previously described protein with mast cell-degranulating properties. MALDI-MS-based de novo sequencing provided a partial amino acid sequence different from that of the previously described allergen Tha p 1, and it was named Tha p 2. This allergen was detected in 61% of patients, and it is therefore a new major caterpillar allergen. CONCLUSIONS: Penetration of the setae from the pine processionary caterpillar delivers their allergenic content in addition to causing mechanical or toxic injury.

Seasonal water stress and the resistance of Pinus yunnanensis to a bark-beetle-associated fungus.

We examined the influence of seasonal water stress on the resistance of Pinus yunnanensis (Franch.) to inoculation with Leptographium yunnanense, a pathogenic fungus associated with the aggressive bark beetle, Tomicus n. sp. Experiments took place between October 1997 and November 1999 in two plots located at the top and at the foot of a hill in Shaogiu, China, a region characterized by dry winters and wet summers. Following isolated and mass fungal inoculations, we observed the reaction zone length, fungal growth in the phloem, and the occlusion, blue-staining and specific hydraulic conductivity of the sapwood. Measurements of soil and needle water contents and predawn needle water potentials confirmed that trees were subject to mild water stress during winter, especially at the drier hilltop site. Measures of tree resistance to fungal infection of phloem and sapwood were congruent and indicated that trees were most susceptible to inoculation during the wet summer, especially at the lower-elevation plot. Specific hydraulic conductivity decreased after inoculation in summer. The results indicate that mild seasonal water stress is not likely responsible for the recent extensive damage to young P. yunnanensis stands by Tomicus n. sp. in the vicinity of our study plots. Rather, the results suggest that mild water stress enhances tree resistance to fungal pathogens associated with Tomicus n. sp.

High intercontinental migration rates and population admixture in the sapstain fungus Ophiostoma ips.

Ophiostoma ips is a common fungal associate of various conifer-infesting bark beetles in their native ranges and has been introduced into non-native pine plantations in the Southern Hemisphere. In this study, we used 10 microsatellite markers to investigate the population biology of O. ips in native (Cuba, France, Morocco and USA) and non-native (Australia, Chile and South Africa) areas to characterize host specificity, reproductive behaviour, and the potential origin as well as patterns of spread of the fungus and its insect vectors. The markers resolved a total of 41 alleles and 75 haplotypes. Higher genetic diversity was found in the native populations than in the introduced populations. Based on the origin of the insect vectors, the populations of O. ips in Australia would be expected to reflect a North American origin, and those in Chile and South Africa to reflect a European origin. However, most alleles observed in the native European population were also found in the native North American population; only the allele frequencies among the populations varied. This admixture made it impossible to confirm the origin of the introduced Southern Hemisphere (SH) populations of O. ips. There was also no evidence for specificity of the fungus to particular bark beetle vectors or hosts. Although O. ips is thought to be mainly self-fertilizing, evidence for recombination was found in the four native populations surveyed. The higher genetic diversity in the North American than in the European population suggests that North America could be the possible source region of O. ips.

Host selection in Tomicus piniperda L.: composition of monoterpene hydrocarbons in relation to attack frequency in the shoot feeding phase.

The aim of this study was to investigate the host selection capacity of the pine shoot beetle, Tomicus piniperda, in the shoot-feeding phase and analyze the chiral and non-chiral host volatiles by means of GC-MS and 2D-GC in five Pinus species originating from France (Pinus sylvestris, P. halepensis, P. nigra laricio, P. pinaster maritima, P. pinaster mesogeensis). Dominating monoterpenes were (-)-alpha-pinene, (+)-alpha-pinene, (-)-beta-pinene and (+)-3-carene. The amounts of the enantiomers varied considerably within and among the species. In a principal component analysis-plot, based on the absolute amounts of 18 monoterpene hydrocarbons, separation of the pine species into two groups was obtained. P. halepensis and P. sylvestris were grouped according to the amount of (+)-alpha-pinene and (+)-3-carene, while P. nigra laricio, P. pinaster maritima and P. pinaster mesogeensis were grouped according to (-)-alpha-pinene and (-)-beta-pinene. P. nigra laricio was the species most attacked and P, halepensis the one least attacked by T. piniperda.

Intra- and inter-provenance variability in phloem phenols of Picea abies and relationship to a bark beetle-associated fungus.

One hundred Norway spruce (Picea abies (L.) Karst.) clones (three ramets per clone) were analyzed for phloem phenol composition and concentration before and 10 days after wound inoculation with sterile malt agar. Fifty clones (Experiment 1) belonged to the same provenance, whereas the remaining clones (Experiment 2) belonged to five provenances from three geographic areas. In Experiment 2, two additional ramets from the same clones were mass inoculated with Ceratocystis polonica (Siem.) C. Moreau to quantify the resistance of each clone. Tree response to wound inoculations was characterized by increased catechin concentration in both experiments, accompanied by increases in astringin and decreases in piceid in Experiment 1. In both experiments, we observed a diverse group of phenolic compounds whose concentrations increased (catechin, astringin) or did not vary (taxifolin glucoside) in response to wound inoculations, whereas concentrations of a homogeneous group of stilbene compounds decreased (piceid) or did not vary (isorhapontin, unidentified stilbene). In Experiment 2, provenances from the alpine and Hercynian-Carpatic areas differed from provenances from the Baltic area with respect to the relative importance of these two groups of compounds, further indicating that the two groupings of phenolic compounds structure the Norway spruce populations. Eighty days after mass inoculation, the percentage of healthy sapwood, which was taken as a measure of tree resistance, indicated that clones from the Baltic area were less resistant to mass inoculations than clones from the alpine and Hercynian-Carpatic areas. We conclude that the degree of resistance of Norway spruce trees to mass inoculations with a bark beetle-associated fungus can be predicted based on the diversity of constitutive phloem phenols and the ability to induce phenol synthesis in response to wounding.