Search

The number of non-native invasive pests and pathogens has increased dramatically in recent years, with disastrous consequences for the health of forests worldwide. Multiple studies have shown that mixed forests may suffer less damage from insect pests than single species forest. This “associational resistance” can be notably explained by the fact that heterospecific neighbours make it more difficult for herbivores to locate and then exploit their host tree. However, the validity of these findings in the case of non-native, invasive pests and pathogens remains to be demonstrated. In this study, we monitored over two hundred Douglas firs in pure and mixed plots of a tree diversity experiment to assess the damage from the non-native gall midge Contarinia pseudotsugae and the non-native needle cast Nothophaeocryptopus gaeumannii. The probability of Swiss needle-cast infection was lower in Douglas fir trees surrounded by heterospecific neighbours. Gall midge damage was lower on Douglas firs surrounded by taller neighbours, consistent with the hypothesis of reduced host Apparency. Douglas fir trees that were more damaged by C. pseudotsugae were also more often infected by N. gaeumannii. Our study thus provides partial support of the associational resistance hypothesis of mixed forests against exotic pests and pathogens. Promoting forest species diversity at the stand level could, therefore, offer interesting prospects for reducing the impact of biological invasions, especially those involving both pests and pathogens.

When a non-native species succeeds in establishing in a new habitat, one of the possible responses is to attempt its eradication. In the present study, we analysed European eradication programmes against non-native pests and pathogens of woody plants (PPWP) from 1945 to date. Our main goal was to identify which factors affect the success of an eradication programme, reinforcing guidelines for future eradication of PPWP. Data on eradication campaigns were obtained from online databases, scientific and grey literature, and Plant Protection Organizations’ reports. Factors influencing eradication success for both arthropods and pathogens were analysed with LASSO regression and decision tree learning. A total of 848 cases officially declared as eradication attempts were documented in our database (8-fold higher than previous reports). Both the number of programmes and their rate of success increased sharply over the last two decades. Only less than 10% of the non-native organisms affecting woody plants were targeted for attempted eradication despite the high economic and ecological impacts caused by some species for which no efforts were undertaken. Almost one-third of the officially declared cases of eradication concerned organisms that were still restricted to the material with which they were introduced. For these cases the success rate was 100%. The success rate of established species was only 50% for arthropods and 61% for pathogens. The spatial extent of the outbreak was the factor that most affected the outcome of eradication campaigns. The eradication success decreased abruptly above 100 ha for arthropods and 10 ha for pathogens. Additionally, other variables were shown to influence the outcome of eradication programmes, in particular the type of environment, with the highest eradication success rate found in nurseries and glasshouses, with successful outcomes increasing if quarantine measures were applied and when monitoring included asymptomatic plants. Particular species traits may reduce eradication success: parthenogenetic arthropods, saprotrophic pathogens, wind dispersal, the possibility to remain asymptomatic indefinitely, and the existence of resting spores or stages. In conclusion, small affected areas, quick response, and efficient implementation of quarantine restrictions, together with particular species traits, may allow a high probability of eradication success. Preparedness at the country and European level would allow a larger number of target species to be included in future eradication programmes.

In a pilot study, a biological control with hypovirus-infected Cryphonectria parasitica was applied in 3 study plots in Turkey, in June 2013. The aim of this study was to evaluate the success of the applied biological control by verifying the establishment of the released hypovirus (CHV-1) strains using molecular markers. C. parasitica isolates were sampled from cankers at 3 different time points: before the biological control treatments (April 2013), and 5 months (October 2013) and 11 months (May 2014) after the treatment. In total, 255 C. parasitica isolates were recovered and characterized. First, the culture morphology and vegetative compatibility type of these isolates were assessed. Next, the presence of hypoviruses in white isolates was checked by RNA extraction and subsequent RTPCR. Finally, a sequence analysis was performed to compare the hypoviruses to the released biological control hypoviruses by examining single nucleotide markers and reconstructing their phylogenetic relationship. The study sites in Izmir and Kutahya were found to be free of hypoviruses prior to the biological control, whereas in Sinop, the occurrence of hypoviruses of the Italian subtype was observed. Reisolations of the treated cankers and subsequent molecular analysis resulted in the detection of the released biocontrol hypovirus strains in all 3 study sites. The reisolated hypoviruses in Izmir and Kutahya originated from either one of the released biocontrol strains. In Sinop, both natural and artificially introduced hypoviruses were detected. Our study showed that the released biological control hypoviruses persisted in the treated cankers, which is promising for the biological control of chestnut blight in Turkey.

Chestnut blight has spread throughout Europe since the introduction of its causal agent, Cryphonectria parasitica, >70 years ago. In our study, we analyzed the diversity of vegetative compatibility (vc) and microsatellite genotypes of C. parasitica, as well as sequence diversity of Cryphonectria hypovirus 1 (CHV1) in six populations from Switzerland, Croatia, and North Macedonia. Resampling of local populations that were already investigated more than a decade ago allowed us to analyze the spatial and temporal population structure across an invasive range of the pathogen in Europe. Regardless of which genetic marker was used, the >60-year-old Swiss and Croatian populations had high population diversity, whereas more recent North Macedonian populations were mostly clonal. These diversity differences between the investigated populations remained stable over time. A high diversity of CHV1 was observed in all three countries, with North Macedonian strains forming a separate cluster from strains obtained in other countries. No correlation between vc diversity and CHV1 prevalence was observed, suggesting a well-established and maintained natural hypovirulence in all countries, further corroborated by an observed increase in genetic diversity of Croatian C. parasitica populations over time, without collapse of CHV1 prevalence.

International audience; The basidiomycete genus Armillaria s.l. (Armillaria s.s. and Desarmillaria) has a worldwide distribution and plays a central role in the dynamics of numerous woody ecosystems, including natural forests, tree plantations for timber production, orchards, vineyards and gardens. Early studies have shown that all Armillaria species are capable of degrading dead woody substrates, causing white rot. Moreover, most species exhibit a parasitic ability, and can be considered as facultative necrotrophs. Although over the years extensive research has been conducted on the phylogeny, biology and ecology of different Armillaria species, numerous theoretical and applied questions remain open. Recently published studies have provided new perspectives, the most significant of which we present in this review. First, new investigations have highlighted the importance of a multilocus approach for depicting the phylogeny of the genus Armillaria. Second, the importance of clonality and sexuality for the different species is now better described, enabling a more accurate prediction of population dynamics in various environments. Third, genome sequencing has provided new insights into genome evolution and the genetic basis of pathogenicity and wood degradation ability. Fourth, several new studies have pointed out the possible influence of climate change on Armillaria distribution, biology and ecology, raising questions regarding the future evolution of Armillaria species and their effect on ecosystems. In this review, we also give a state-of-the-art overview of the control possibilities of parasitic Armillaria species. Finally, we outline some still open questions in Armillaria research, the investigation of which will strongly benefit from recent methodological advances.

Invasions by non-native pathogens represent a major threat to managed and natural ecosystems worldwide. Although necessary for adopting preventive strategies, the identification of invasive species before they are introduced is particularly difficult. Indeed, most pathogenic species that have become established in the last decades were first described only after they became invasive. To prevent further biological invasions, not only the early identification of potential new invasive plant pathogens is crucial, but also the assessment of their potential host range. In this study, we determined the pathogenicity and the saprotrophic ability of three Cryphonectria species towards three potential hosts in the family Fagaceae. For this, seedlings and dormant stems of European chestnut (Castanea sativa), pedunculate oak (Quercus robur) and European beech (Fagus sylvatica) were inoculated with different genotypes of C. parasitica (Asian species, invasive in Europe), C. naterciae (European species), and C. japonica (Asian species, not present in Europe). Lesion growth was measured and mortality assessed for four months. The highest damage was caused by C. parasitica on European chestnut, while C. japonica and C. naterciae induced significantly smaller lesions on this host species. All three Cryphonectria species did not grow saprophytically on F. sylvatica and Q. robur, but successfully colonized dormant stems of C. sativa. In the context of biological invasions, our study shows that the Asian C. japonica most likely represents a much less severe threat than C. parasitica for the tested European host species. Nonetheless, the ability of C. naterciae and C. japonica to saprotrophically colonize fresh chestnut wood may suggest that they could become established in chestnut forests and eventually infect weakened chestnut trees or other hosts not tested in this study.

International audience; Gnomoniopsis castaneae is an emerging fungal pathogen currently scored as the major nut rot agent on chestnut, although it is also associated with cankers on both chestnut and hazelnut, as well as with necrosis on chestnut galls and leaves. Described for the first time in 2012, G. castaneae has been reported in several countries across Europe, Asia and Australasia, often in relation to severe outbreaks. The goal of this review is to provide a comprehensive summary of the state of the art about G. castaneae, highlighting the main results achieved by the research and stressing the most relevant knowledge gaps that still need to be filled. This overview includes topics encompassing the taxonomy of the fungal pathogen, its host range and geographic distribution, the symptomatology and the diagnostic methods available for its detection, its impact, biology, ecology and epidemiology. The main interactions between G. castaneae and other organisms are also discussed, as well as the possible control strategies. In these past few years, relevant progresses in the knowledge of G. castaneae have been achieved, yet the complexity of the challenges that this pathogen poses to chestnut growers and to the scientific community advocates for further advances.

Aim Early warning against potentially harmful organisms of woody plant species can be achieved by sampling sentinel plants in exporting countries. However, it is unclear where sentinel plants can best be located, and how many samples are required and when and how often sampling optimally should take place for the adequate assessment of the biodiversity associated with the target plant species. We aimed to review spatial and temporal factors affecting associate biodiversity of single woody plant species and to develop guidance for the design of global biodiversity sampling studies. Location Worldwide. Taxon Insects and Fungi. Methods Literature about factors affecting the diversity of insects and fungi in association with single plant species on global, regional, local and different temporal scales was reviewed. Case studies of insect and fungal diversity, primarily collected on single plant species, and the cost of collecting and analysing samples from locations around the world were analysed. Results The review of the literature illustrated various factors affecting diversity, and the case studies allowed quantification of the relative impact of some spatial, temporal and financial aspects on captured biodiversity and, thus, illustrate the need to consider all possible factors that may affect the result of the sampling when deciding on a sampling design. Main conclusions Our study illustrates the factors that should be considered when deciding on the location and timing of sampling for sentinel plants, which is important because of the trade‐off between the number of samples and sampling locations needed to detect many of the species which may be potential pests, and the cost of (repeated) sampling in many locations. Decisions about the sampling design must be based on the objective of the sampling, but our recommendations apply irrespective of the targeted plant species or country.

International audience; The international seed trade is considered relatively safe from a phytosanitary point of view and is therefore less regulated than trade in other plants for planting. However, the pests carried by traded seeds are not well known. We assessed insects and fungi in 58 traded seed lots of 11 gymnosperm and angiosperm tree species from North America, Europe, and Asia. Insects were detected by X-raying and molecular methods. The fungal community was characterized using high-throughput sequencing (HTS) and by growing fungi on non-selective agar. About 30% of the seed lots contained insect larvae. Gymnosperms contained mostly hymenopteran (Megastigmus spp.) and dipteran (Cecidomyiidae) larvae, while angiosperms contained lepidopteran (Cydia latiferreana) and coleopteran (Curculio spp.) larvae. HTS indicated the presence of fungi in all seed lots and fungi grew on non-selective agar from 96% of the seed lots. Fungal abundance and diversity were much higher than insect diversity and abundance, especially in angiosperm seeds. Almost 50% of all fungal exact sequence variants (ESVs) found in angiosperms were potential pathogens, in comparison with around 30% of potentially pathogenic ESVs found in gymnosperms. The results of this study indicate that seeds may pose a greater risk of pest introduction than previously believed or accounted for. A rapid risk assessment suggests that only a small number of species identified in this study is of phytosanitary concern. However, more research is needed to enable better risk assessment, especially to increase knowledge about the potential for transmission of fungi to seedlings and the host range and impact of identified species.

International audience; The number of invasive alien pest and pathogen species affecting ecosystem functioning, human health and economies has increased dramatically over the last decades. Discoveries of invasive pests and pathogens previously unknown to science or with unknown host associations yet damaging on novel hosts highlights the necessity of developing novel tools to predict their appearance in hitherto naive environments. The use of sentinel plant systems is a promising tool to improve the detection of pests and pathogens before introduction and to provide valuable information for the development of preventative measures to minimize economic or environmental impacts. Though sentinel plantings have been established and studied during the last decade, there still remains a great need for guidance on which tools and protocols to put into practice in order to make assessments accurate and reliable. The sampling and diagnostic protocols chosen should enable as much information as possible about potential damaging agents and species identification. Consistency and comparison of results are based on the adoption of common procedures for sampling design and sample processing. In this paper, we suggest harmonized procedures that should be used in sentinel planting surveys for effective sampling and identification of potential pests and pathogens. We also review the benefits and limitations of various diagnostic methods for early detection in sentinel systems, and the feasibility of the results obtained supporting National Plant Protection Organizations in pest and commodity risk analysis.

The xylophagous cerambycid Anoplophora glabripennis, the Asian long-horned beetle (ALB), is highly polyphagous and can colonize a wide range of broadleaved host trees causing significant economic damage. For this reason, it is considered a quarantine pest in Europe and North America. Although the global spread of ALB has been depicted recently, no comprehensive studies exist on the genetic pattern of populations’ establishment and dynamics at fine-scale (i.e. within invasive outbreaks), before eradication measures are applied. This information may, however, be particularly important for an efficient management and control of invasive pests. Here, we characterized population genetic diversity and patterns of spread of ALB within and among the four outbreaks detected in Switzerland between 2011 and 2015. For this, we genotyped 223 specimens at 15 nuclear microsatellite loci and conducted specific population-based analyses. Our study shows: (1) At least three independent introductions and a, human-mediated, secondary dispersal event leading to the four outbreaks in the country; (2) An overall low intra-population genetic diversity in the viable and several years active invasive populations; (3) A colonization of single trees by homogeneous ALB genotypes; And (4) an establishment of populations several generations prior to its official discovery.

International audience; Retracing introduction routes is crucial for understanding the evolutionary processes involved in an invasion, as well as for highlighting the invasion history of a species at the global scale. The Asian long-horned beetle (ALB) Anoplophora glabripennis is a xylophagous pest native to Asia and invasive in North America and Europe. It is responsible for severe losses of urban trees, in both its native and invaded ranges. Based on historical and genetic data, several hypotheses have been formulated concerning its invasion history, including the possibility of multiple introductions from the native zone and secondary dispersal within the invaded areas, but none have been formally tested. In this study, we characterized the genetic structure of ALB in both its native and invaded ranges using microsatellites. In order to test different invasion scenarios, we used an approximate Bayesian "random forest" algorithm together with traditional population genetics approaches. The strong population differentiation observed in the native area was not geographically structured, suggesting complex migration events that were probably human-mediated. Both native and invasive populations had low genetic diversity, but this characteristic did not prevent the success of the ALB invasions. Our results highlight the complexity of invasion pathways for insect pests. Specifically, our findings indicate that invasive species might be repeatedly introduced from their native range, and they emphasize the importance of multiple, human-mediated introductions in successful invasions. Finally, our results demonstrate that invasive species can spread across continents following a bridgehead path, in which an invasive population may have acted as a source for another invasion.

Infection of American and European chestnuts with the chestnut blight fungus Cryphonectria parasitica results in the formation of cankers, lesions caused by the growth of mycelia within bark tissue of the host plant. Infection of the fungus with Cryphonectria hypovirus 1 (CHV‐1) results in conversion of the mycelial phenotype from virulent to hypovirulent, thus allowing production of callus around cankers as a reaction by infected trees, rendering active into inactive cankers. In this study, we sampled one USA and six European chestnut stands and assessed frequency of hypovirulent C. parasitica and diversity of vegetative compatibility (vc) types present in calluses and randomly sampled cankers. Callused cankers on C. dentata at West Salem in the USA yielded significantly more hypovirulent C. parasitica isolates compared with four sampled populations on C. sativa, while all six sampled European populations did not show any statistically significant differences among themselves. We observed no correlation between hypovirulence frequencies in randomly sampled cankers and calluses, as well as no correlation of C. parasitica vc type diversity in calluses and residential populations of the fungus. Furthermore, even though we have observed calluses with more than one vc type, they do not occur regularly. Even when present in C. parasitica populations with high vc type diversity, no more than three different vc types were observed in a single callus.

In this study, we investigated the diversity and ecology of Armillaria species in virgin pure beech and mixed conifer forests (15,000 ha) of the Carpathian Biosphere Reserve in Ukraine. Armillaria rhizomorphs were systematically sampled, both from the soil and from the root collar of trees (epiphytic), on 79 plots (25 × 20 m) of a 1.5 × 1.5 km grid. In both forest massifs, rhizomorphs were present in the majority of the soil samples, with an estimated dry weight of 512 kg/ha in the pure beech forests and 223 kg/ha in the mixed conifer forests. Similarly, in both forest massifs, most of the trees inspected had rhizomorphs at the root collar. Species identification based on DNA analyses showed that all five annulated European Armillaria species occur in these virgin forests, as previously observed in managed forests in central Europe. However, differences in the frequencies of the single species were observed. The predominance of the preferentially saprotrophic A. cepistipes and A. gallica (84 and 15% of the specimens, respectively) and the absence of significant pathogenic activity suggest that in these virgin forests Armillaria species are most likely to behave as saprotrophs. Forest management may increase the frequency of the pathogenic species A. ostoyae, which is rare in virgin forests.

Introduced plant pathogens are increasingly recognized as a major threat to biodiversity and ecosystem functioning. One such pathogen, the causal agent of the devastating ash dieback in Europe, Hymenoscyphus fraxineus, was most likely introduced into Europe from eastern Asia in the 1990s. To investigate the genetic population structure of this invasive fungus at the epidemic disease front (Switzerland) and in the post-epidemic phase (Lithuania), a total of 847 H. fraxineus isolates were genotyped at 11 microsatellite loci. Among these isolates, 244 multilocus genotypes were found in five post-epidemic subpopulations (367 isolates) of the fungus and 263 in five epidemic subpopulations (480 isolates). No genetic differentiation was found between isolates recovered from bark lesions and fallen leaf petioles, which suggests that all H. fraxineus genotypes have the potential to induce bark infections on living trees and to survive saprophytically. Moreover, no genetic differentiation and no difference in genetic diversity were detected between the epidemic and post-epidemic populations. The entire genetic diversity present in the original founding populations in north-eastern Europe seems to have been transmitted to the epidemic disease front. Nonetheless, gene flow among post-epidemic subpopulations occurs slightly more random than among epidemic subpopulations. Furthermore, the probability of correct assignment of a particular H. fraxineus genotype to its subpopulation of origin was greater in Switzerland than in Lithuania. These two analyses point to weak founder effects at the disease front.

Armillaria possesses several intriguing characteristics that have inspired wide interest in understanding phylogenetic relationships within and among species of this genus. Nuclear ribosomal DNA sequence-based analyses of Armillaria provide only limited information for phylogenetic studies among widely divergent taxa. More recent studies have shown that translation elongation factor 1-α (tef1) sequences are highly informative for phylogenetic analysis of Armillaria species within diverse global regions. This study used Neighbor-net and coalescence-based Bayesian analyses to examine phylogenetic relationships of newly determined and existing tef1 sequences derived from diverse Armillaria species from across the Northern Hemisphere, with Southern Hemisphere Armillaria species included for reference. Based on the Bayesian analysis of tef1 sequences, Armillaria species from the Northern Hemisphere are generally contained within the following four superclades, which are named according to the specific epithet of the most frequently cited species within the superclade: (i) Socialis/Tabescens (exannulate) superclade including Eurasian A. ectypa, North American A. socialis (A. tabescens), and Eurasian A. socialis (A. tabescens) clades; (ii) Mellea superclade including undescribed annulate North American Armillaria sp. (Mexico) and four separate clades of A. mellea (Europe and Iran, eastern Asia, and two groups from North America); (iii) Gallica superclade including Armillaria Nag E (Japan), multiple clades of A. gallica (Asia and Europe), A. calvescens (eastern North America), A. cepistipes (North America), A. altimontana (western USA), A. nabsnona (North America and Japan), and at least two A. gallica clades (North America); and (iv) Solidipes/Ostoyae superclade including two A. solidipes/ostoyae clades (North America), A. gemina (eastern USA), A. solidipes/ostoyae (Eurasia), A. cepistipes (Europe and Japan), A. sinapina (North America and Japan), and A. borealis (Eurasia) clade 2. Of note is that A. borealis (Eurasia) clade 1 appears basal to the Solidipes/Ostoyae and Gallica superclades. The Neighbor-net analysis showed similar phylogenetic relationships. This study further demonstrates the utility of tef1 for global phylogenetic studies of Armillaria species and provides critical insights into multiple taxonomic issues that warrant further study.

Invasive oomycete pathogens have been causing significant damage to native ecosystems worldwide for over a century. A recent well-known example is Phytophthora ramorum, the causal agent of sudden oak death, which emerged in the 1990s in Europe and North America. In Europe, this pathogen is mainly restricted to woody ornamentals in nurseries and public greens, while severe outbreaks in the wild have only been reported in the UK. This study presents the results of the P. ramorum survey conducted in Swiss nurseries between 2003 and 2011. In all 120 nurseries subjected to the plant passport system, the main P. ramorum hosts were visually checked for above ground infections. Phytophthora species were isolated from tissue showing symptoms and identified on the basis of the morphological features of the cultures and sequencing of the ribosomal ITS region. Phytophthora was detected on 125 plants (66 Viburnum, 58 Rhododendron and one Pieris). Phytophthora ramorum was the most frequent species (59·2% of the plants), followed by P. plurivora, P. cactorum, P. citrophthora, P. cinnamomi, P. cactorum/P. hedraiandra, P. multivora and P. taxon PgChlamydo. The highest incidence of P. ramorum was observed on Viburnum × bodnantense. Microsatellite genotyping showed that the Swiss P. ramorum population is highly clonal and consists of seven genotypes (five previously reported in Europe, two new), all belonging to the European EU1 clonal lineage. It can therefore be assumed that P. ramorum entered Switzerland through nursery trade. Despite sanitation measures, repeated P. ramorum infections have been recorded in seven nurseries, suggesting either reintroduction or unsuccessful eradication efforts.

Phytophthora ramorum, the causal agent of sudden oak death and ramorum blight, is known to exist as three distinct clonal lineages which can only be distinguished by performing molecular marker-based analyses. However, in the recent literature there exists no consensus on naming of these lineages. Here we propose a system for naming clonal lineages of P. ramorum based on a consensus established by the P. ramorum research community. Clonal lineages are named with a two letter identifier for the continent on which they were first found (e.g., NA = North America; EU = Europe) followed by a number indicating order of appearance. Clonal lineages known to date are designated NA1 (mating type: A2; distribution: North America; environment: forest and nurseries), NA2 (A2; North America; nurseries), and EU1 (predominantly A1, rarely A2; Europe and North America; nurseries and gardens). It is expected that novel lineages or new variants within the existing three clonal lineages could in time emerge.

We followed the local intensification and dispersal of Phytophthora ramorum Werres, De Cock, Man In't Veld in Oregon tanoak (Lithocarpus densiflorus (Hook Arn.) Rehd.) forests from its initial detection in 2001 through 2006, co- incident with a continuing eradication effort. The initial infested area included nine scattered sites below 400 m elevation, close to the Pacific Ocean near Brookings, Oregon. In subsequent years, one-half of new infections were within 122 m of a previous infection, and 79% of the newly detected trees occurred within 300 m of a previously identified tree. Dispersal up to 4 km was occasionally recorded. Initial infection occurred in the upper crowns of tanoak trees. The pathogen was re- covered in rainwater collected beneath diseased tanoak trees in every month from November 2006 through October 2007. Twenty-four multilocus microsatellite genotypes were identified among 272 P. ramorum isolates collected from Curry County. Genotypic analysis provided independent estimates of time of origin of the Oregon infestation, its clustered distri- bution, and dispersal distances. In all sampling years, 60%-71% of the isolates belonged to the same multilocus genotype. In 2001, 12 genotypes were detected and new genotypes were identified in each of the subsequent years, but all isolates belonged to the same clonal lineage. Knowledge of local intensification of the disease and long-distance dispersal should inform both Oregon eradication efforts and national quarantine regulations. Resume´ : Nous avons suivi la progression locale et la dispersion de Phytophtora ramorum Werres, De Cock, Man In't Veld dans des forets de lithocarpe de Californie (Lithocarpus densiflorus (Hook. Arn.) Rehd.) en Oregon depuis que la maladie a etedetectee pour la premiere fois, en 2001, jusqu'en 2006, pendant que se poursuivait les efforts d'eradication. Les zones initialement infectees incluaient neuf stations dispersees en dessous de 400 m d'altitude, aproximitede l'ocean Pacifique, pres de Brookings en Oregon. Au cours des annees qui ont suivi, la moitiedes nouvelles infections etaient sit- uees amoins de 122 m d'une infection precedente et 79% des arbres nouvellement infectesetaient situesamoins de 300 m d'un arbre dejainfecte´. La dispersion de la maladie a occasionnellement eteobservee sur une distance pouvant aller jus- qu'aquatre kilometres. L'infection initiale s'est produite dans la partie superieure de la cime des lithocarpes. Le cham- pignon pathogene a eteretrouvedans l'eau de pluie collectee sous les lithocarpes malades achaque mois, de novembre 2006 aoctobre 2007. Vingt-quatre genotypes multilocus de microsatellites ont eteidentifies parmi les 272 isolats de P. ra- morum collectes dans le comtede Curry. L'analyse genotypique a fourni des estimations independantes du moment du de´- but de l'infection en Oregon, de sa repartition en grappes et des distances de dispersion. Durant toutes les annees

The genetic structure of the clonally reproducing Sudden Oak Death (SOD) pathogen in California was investigated using seven variable microsatellites. A total of 35 multilocus genotypes were identified among 292 samples representative of populations from 14 forest sites and of the nursery trade. amova indicated significant genetic variability both within (44.34%) and among populations (55.66%). Spatial autocorrelation analyses indicated that Moran's index of similarity reached a minimum of 0.1 at 350 m, increased to 0.4 at 1500 m and then decreased to zero at 10 km. These results suggest a bimodal pattern of spread, with medium range dispersal (1500-10,000 m) putatively attributed to the presence of strong winds. Lack of genetic structure was identified for three groups of populations. One group notably included the nurseries' population and two forest populations, both linked to early reports of the pathogen. A neighbour-joining analysis based on pairwise Phi(ST) values indicated that the clade inclusive of the nurseries' populations is basal to all California populations. A network analysis identified three common genotypes as the likely founders of the California infestation and proposes a stepwise model for local evolution of novel genotypes. This was supported by the identification in the same locations of novel genotypes and of their 1- or 2-step parents. We hypothesize that the few undifferentiated population groups indicate historical human spread of the pathogen, while the general presence of genetically structured populations indicates that new infestations are currently generated by rare medium or long-range natural movement of the pathogen, followed by local generation of new genotypes.

[EN] An analysis of incidence of Phytophthora spp. in 732 European nurseries producing forest transplants, larger specimen trees, landscape plants and ornamentals, plus 2525 areas in which trees and shrubs were planted, is presented based on work conducted by 38 research groups in 23 European countries between 1972 and 2013. Forty-nine Phytophthora taxa were recorded in 670 nurseries (91.5%); within these nurseries, 1614 of 1992 nursery stands (81.0%) were infested, although most affected plants appeared healthy. In forest and landscape plantings, 56 Phytophthora taxa were recovered from 1667 of 2525 tested sites (66.0%). Affected plants frequently showed symptoms such as crown thinning, chlorosis and dieback caused by extensive fine root losses and/or collar rot. Many well-known highly damaging host¿Phytophthora combinations were frequently detected but 297 and 407 new Phytophthora¿host associations were also observed in nurseries and plantings, respectively. On average, 1.3 Phytophthora species/taxa per infested nursery stand and planting site were isolated. At least 47 of the 68 Phytophthora species/taxa detected in nurseries and plantings were exotic species several of which are considered well established in both nurseries and plantings in Europe. Seven known Phytophthora species/taxa were found for the first time in Europe, while 10 taxa had not been previously recorded from nurseries or plantings; in addition, 5 taxa were first detections on woody plant species. Seven Phytophthora taxa were previously unknown to science. The reasons for these failures of plant biosecurity in Europe, implications for forest and semi-natural ecosystems and possible ways to improve biosecurity are discused., The authors are grateful to all public and private owners of nurseries and plantings who contributed to this extensive study. T. Jung acknowledges the support by the Regione Autonoma della Sardegna, Visiting Professor Program at the University of Sassari, Italy. The authors also thank the COST Office and the European Council for providing the European COST Actions FP0801 (http://www.cost.eu/domains_actions/fps/Actions/FP0801) and FP 1002 (http://www.cost.eu/domains_actions/fps/Actions/FP1002), the EU projects FOR-THREATS and ISEFOR, and BiodivERsA project RESIPATH as platforms for stimulating discussions on the nursery pathway and possible management solutions.

International audience; Development of molecular markers for phylogenetic, population genetics and phylogeographic studies remains arduous in non-model species with low or no genomic resources. Sequencing the whole or a large part of the genome of the target species using next-generation sequencing technologies is considered a promising method, although it still needs a large investment in bioinformatics. To quickly find polymorphic markers in fungal species, we tested an alternative method, named PHYLORPH. This method allows users to quickly target polymorphic regions of single copy genes in fungi using public databases. We applied this method to Armillaria species, which are important fungal pathogens and saprophytes playing a central role in the dynamics of forest ecosystems worldwide. We isolated 32 single copy genes with numerous single nucleotide polymorphism (SNP) sites. A genetic analysis of two French populations validated the polymorphism of 80 among 92 SNPs tested, and seven of these sequences exactly reconstructed the known phylogenetic tree of four tested Armillaria species. These results confirmed that the PHYLORPH method is efficient to identify various markers at both the intra- and interspecific levels for fungal species with no or few previous genetic markers.

The impact of invasive fungal pathogens and pests on trees is often studied individually, thereby omitting possible interactions. In this study the ecological interaction between the chestnut blight fungus Cryphonectria parasitica and the chestnut gall wasp Dryocosmus kuriphilus was investigated. We determined if abandoned galls could be colonized by C. parasitica and thereby act as an entry point and a source of pathogen inoculum. Moreover we assessed the identity and diversity of other gall-colonizing fungal species. A total of 1973 galls were randomly sampled from 200 chestnut trees in eight Swiss stands. In a stand C. parasitica was isolated from 0.4–19.2% of the galls. The incidence of C. parasitica on the galls and the fungal diversity significantly increased with the residence time of D. kuriphilus in a stand. All but one C. parasitica cultures were virulent. The predominant fungus isolated from galls was Gnomoniopsis castanea whose abundance influenced negatively that of C. parasitica . This study shows that D. kuriphilus galls can be colonized by virulent strains of the chestnut blight fungus C. parasitica . This can have effects on the chestnut blight incidence even in chestnut stands where the disease is successfully controlled by hypovirulence. The gall wasp presence influences also the fungal species composition on chestnut trees.

Summary In managed spruce forests, Armillaria cepistipes and A. ostoyae are efficient stump colonizers and may compete for these resources when they co-occur at the same site. The aim of this experiment was to quantify the mutual competitive ability of the two Armillaria species in producing rhizomorphs and in colonizing Norway spruce (Picea abies) stumps. Five isolates of A. cepistipes and two isolates of A. ostoyae were simultaneously inoculated pair-wise into pots containing a 4-year-old spruce seedling. For comparison, each isolate was also inoculated alone. One year after inoculation, stumps were created by cutting down the seedlings. Six months after creation of the stumps, rhizomorph production and stump colonization were assessed. Armillaria spp. were identified from 347 rhizomorphs and 48 colonized stumps. Armillaria cepistipes dominated both as rhizomorphs in the soil and on the stumps. Nevertheless, A. ostoyae was relatively more frequent on the stumps than in the soil and A. cepistipes was relatively more frequent in the soil than on the stumps. In both species, the ability to colonize the stumps in simultaneous inoculations was significantly reduced compared with single inoculations. In respect to rhizomorph production, simultaneous co-inoculations had a slightly stimulatory effect on A. cepistipes and no significant effect on A. ostoyae. Our study suggests a rather neutralistic co-existence of A. cepistipes and A. ostoyae as rhizomorphs in the soil. Concerning the ability to colonize stumps, the two species experience a mutual negative effect from the interaction, probably because of interspecific competition. Resume Dans les forets gerees d’epiceas, Armillaria cepistipes et Armillaria ostoyae sont de bons colonisateurs de souches et sont en competition pour cette ressource quand ils sont presents ensemble dans le meme site. L'objectif de l'experimentation etait de quantifier l'aptitude competitive mutuelle des deux especes d’Armillaria pour la production de rhizomorphes et la colonisation de souches d’epicea commun (Picea abies). Cinq isolats de A. cepistipes et deux isolats de A. ostoyae ont ete inocules simultanement par paires dans des pots contenant chacun un semis d’epicea de 4 ans. Chaque isolat a egalement ete inocule seul, a titre de comparaison. Un an apres inoculation, les semis ont ete coupes pour creer des souches. Six mois apres la coupe, la production de rhizomorphes et la coloniasation des souches ont ete estimees. Armillaria spp. a ete identifiea partir de 347 rhizomorphes et 48 souches colonisees. A. cepistipes domine a la fois sous forme de rhizomorphes dans le sol et dans les souches. Toutefois, A. ostoyae est relativement plus frequent sur souches que dans le sol alors que l'inverse est observe pour A. cepistipes. Pour les deux especes, l'aptitude a coloniser les souches a significativement ete reduite dans le cas des inoculations simultanees par rapport aux inoculations simples. En ce qui concerne la production de rhizomorphes, les inoculations simultanees ont eu un leger effet stimulant sur A. cepistipes et pas d'effet significatif sur A. ostoyae. Nos resultats suggerent une co-existence relativement neutre entre A. cepistipes et A. ostoyaea l’etat de rhizomorphes dans le sol. Concernant l‘aptitude a coloniser les souches, les deux especes subissent un effet negatif mutuel de l'interaction, provenant probablement de la competition interspecifique. Zusammenfassung In bewirtschafteten Fichtenwaldern sind Armillaria cepistipes und Armillaria ostoyae effiziente Besiedler von Baumstumpfen und sie konnen um diese Ressource miteinander konkurrieren, da sie haufig gemeinsam vorkommen. Es wurde ein Experiment durchgefuhrt mit dem Ziel, die wechselseitige Konkurrenzfahigkeit dieser beiden Armillariaarten bezuglich ihrer Rhizomorphenproduktion und ihrer Fahigkeit zur Besiedlung von Fichtenstumpfen (Picea abies) zu quantifizieren. Funf Isolate von A. cepistipes und zwei Isolate von A. ostoyae wurden gleichzeitig paarweise in Topfe mit einem vierjahrigen Fichtensamling inokuliert. Zum Vergleich wurde jedes Isolat auch alleine inokuliert. Ein Jahr nach der Inokulation wurden durch Abschneiden der Pflanzen Stumpfe erzeugt. Sechs Monate nach dem Schnitt wurde die Rhizomorphenproduktion und die Besiedelung der Stumpfe erfasst. Die beiden Arten wurden aus 347 Rhizomorphen und 48 besiedelten Stumpfen reisoliert. Armillaria cepistipes dominierte sowohl in Form von Rhizomorphen im Boden als auch in den Stumpfen. Armillaria ostoyae kam haufiger in den Stumpfen vor als im Boden und A. cepistipes wurde haufiger im Boden gefunden als in den Stumpfen. Bei beiden Arten war die Fahigkeit zur Stumpfbesiedelung bei gleichzeitiger Inokulation im Vergleich zur Einzelinokulation signifikant reduziert. Die gleichzeitige Inokulation mit beiden Arten stimulierte die Rhizomorphenproduktion bei A. cepistipes leicht, hatte aber keinen signifikanten Einfluss auf A. ostoyae. Diese Befunde deuten auf eine weitgehend neutrale Koexistenz der Rhizomorphen von A. cepistipes und A. ostoyae im Boden hin, bezuglich ihrer Fahigkeit zur Stumpfbesiedelung hemmen sich beide Arten aber gegenseitig. Dieser Effekt ist wahrscheinlich durch interspezifische Konkurrenz zu erklaren.

Summary The basidiomycetes Armillaria cepistipes and Armillaria ostoyae frequently occur in the same forest stand. In this study, we determined the virulence of 20 isolates of A. cepistipes and 16 isolates of A. ostoyae on four different provenances of 2-year-old Norway spruce (Picea abies). Within 30 months after inoculation, 1.1 and 19.1% of the seedlings inoculated with A. cepistipes and A. ostoyae, respectively, had died or were dying. The incidence of dead and dying seedlings varied between 3 and 49% among the A. ostoyae isolates. The virulence of an isolate was positively correlated to its ability to produce rhizomorphs. One Norway spruce provenance showed significantly lower susceptibility to A. ostoyae than the other three. Rhizomorphs of both Armillaria species were attached to the root surface. The attached rhizomorphs of A. ostoyae, however, were associated with significantly more lesions. The virulence of the isolates was not correlated with their wood-degrading capability for either of the Armillaria species. Resume Les basidiomycetes Armillaria cepistipes et Armillaria ostoyae sont frequemment trouves ensemble dans une meme parcelle forestiere. Dans cette etude, nous avons compare la virulence de 20 isolats de A. cepistipes et 16 isolats d'A. ostoyae sur quatre provenances d'epicea commun (Picea abies). Trente mois apres l'inoculation, 1.1% et 19.1% des semis inocules avec A. cepistipes et A. ostoyae respectivement, etaient morts ou fortement deperissants. Un taux de mortalite de semis (y compris fortement deperissants) etait entre 3% et 49% a ete obtenu selon les isolats d’A. ostoyae. La virulence d'un isolat etait positivement correlee a sa capacite de production de rhizomorphes. Une des provenances d'epicea commun a montre une sensibilite significativement plus faible aA. ostoyae que celle des trois autres provenances. Les rhizomorphes des deux especes d'Armillaire etaient attaches a la surface de la racine. Toutefois, les rhizomorphes attaches d’A. ostoyaeetaient significativement plus souvent associes a des lesions. Pour les deux especes d'Armillaire, la virulence des isolats n'etait pas correlee avec leur capacite de degradation du bois. Zusammenfassung Die beiden Hallimasch-Arten Armillaria cepistipes und A. ostoyae kommen haufig zusammen im gleichen Waldbestand vor. In der vorliegenden Arbeit wurde die Virulenz von 20 Isolaten von A. cepistipes und 16 Isolaten von A. ostoyae an zweijahrigen Fichtensamlingen (Picea abies) aus vier verschiedenen Provenienzen untersucht. Nach 30 Monaten waren 1.1% (22 von 1980) der Samlinge als Folge einer Infektion durch A. cepistipes abgestorben oder am absterben. Bei A. ostoyae waren im gleiche Zeitraum 19.1% (255 von 1335) der Samlinge betroffen, wobei eine grosse Variation (3–49%) zwischen den Pilzisolaten festgestellt wurde. Die A. ostoyae Isolate mit der hochsten Virulenz zeigten auch die starkste Rhizomorphen-Bildung. Eine Fichten-Provenienz war signifikant weniger anfallig gegen A. ostoyae als die drei anderen Provenienzen. Rhizomorphen beider Hallimasch-Arten bildeten ahnlich haufig feste Kontaktstellen an den Wurzeln der Samlinge. Die Rhizomorphen von A. ostoyae verursachten jedoch signifikant haufiger Wurzellasionen. Bei beiden Hallimasch-Arten wurde kein Zusammenhang zwischen der Virulenz und der Fahigkeit Holz abzubauen gefunden.

Summary • The preferentially saprotrophic basidiomycete Armillaria cepistipes and the pathogenic Armillaria ostoyae occur sympatrically in many European forests. • The spatial population structure of both Armillaria species was investigated in three 1-ha plots established in managed Norway spruce (Picea abies) forests in the Swiss Alps (1400 m asl). A total of 740 Armillaria isolates, 296 from rhizomorphs in the soil and 444 from the stumps, were recovered and identified to species and genets (somatic incompatibility groups). • The incidence of A. cepistipes and A. ostoyae varied greatly among the plots. Two to six genets of each Armillaria species were identified and mapped in each plot. Genets of the same species overlapped rarely and only on the borders. Large spatial overlaps between A. cepistipes and A. ostoyae genets were observed in the plot with the highest incidence of fragmented genets. In five subplots (c. 0.1 ha) established along interspecific boundaries, overlaps were found in all three plots. • Our study suggests a strong intraspecific competition in both Armillaria species. Evidence for competition between the two species, as indicated by spatial mutual exclusion, was only observed on parts of the investigated area.

The ability of the preferentially saprotrophic fungus Armillaria cepistipes and the pathogenic A. ostoyae to capture fresh stump resources was investigated in managed Alpine Norway spruce (Picea abies) forests where both species occur sympatrically. The incidence of Armillaria species and genets as rhizomorphs in the soil as well as external and internal (heart rot) stump colonisation were determined in three comparable 1 ha plots. The results indicate that A. cepistipes and A. ostoyae have a very similar strategy to capture fresh stump resources. Both species produce dense networks of rhizomorphs in the soil enveloping living trees. After the felling of the trees, the stumps are rapidly captured through spread of mycelial fans in the cambial zone. Our study suggests that A. ostoyae is slightly more competitive than A. cepistipes in primary resource capture.

Summary Chestnut blight was first recorded in Azerbaijan on the native European chestnut (Castanea sativa) in 2004, and since then, the disease is expanding in the country. In this study, chestnut blight was detected in seven of eight forest districts that were inspected. To characterize the local population structure of the chestnut blight fungus, 199 Cryphonectria parasitica isolates were collected and assessed for vc type, mating type and microsatellite genotype. Among these isolates, one dominant vc type was detected comprising 93% of the isolates. Six additional vc types were identified among the other isolates. The microsatellite analysis revealed a very similar pattern with 96.5% of the isolates belonging to the same multilocus genotype. Both mating types of C. parasitica were present in all seven districts with a mating type ratio not different from 1:1 in five districts. In accordance with the occurrence of both mating types, sexual fruiting bodies (perithecia) of C. parasitica were found in all districts with an overall prevalence on 20.6% of the cankers. The dominant vc type and microsatellite genotype in Azerbaijan do not occur in Europe, but have previously been found to be widespread in neighbouring Georgia. Our study reveals that sexual reproduction in the invasive C. parasitica population in Azerbaijan is frequent, although the population shows a low genetic diversity. This could favour the biological control of chestnut blight using hypovirulence, which so far does not seem to be present in Azerbaijan.

Human activity has been shown to considerably affect the spread of dangerous pests and pathogens worldwide. Therefore, strict regulations of international trade exist for particularly harmful pathogenic organisms. Phytophthora plurivora, which is not subject to regulations, is a plant pathogen frequently found on a broad range of host species, both in natural and artificial environments. It is supposed to be native to Europe while resident populations are also present in the US. We characterized a hierarchical sample of isolates from Europe and the US and conducted coalescent-, migration, and population genetic analysis of sequence and microsatellite data, to determine the pathways of spread and the demographic history of this pathogen. We found P. plurivora populations to be moderately diverse but not geographically structured. High levels of gene flow were observed within Europe and unidirectional from Europe to the US. Coalescent analyses revealed a signal of a recent expansion of the global P. plurivora population. Our study shows that P. plurivora has most likely been spread around the world by nursery trade of diseased plant material. In particular, P. plurivora was introduced into the US from Europe. International trade has allowed the pathogen to colonize new environments and/or hosts, resulting in population growth.

We describe specific primers and conditions to amplify two dinucleotide and five trinucleotide microsatellite DNA loci isolated from the oomycete Phytophthora ramorum, the causal agent of sudden oak death. The primer sets were tested on 14–30 isolates from North America and Europe. Seven of 14 loci differentiated between A1 and A2 mating types. All seven loci successfully amplified DNA isolated from infected plant tissue. Four loci may be useful for the diagnosis of P. ramorum because they do not amplify closely related Phytophthora species.

The recently emerged plant pathogen Phytophthora ramorum is responsible for causing the sudden oak death epidemic. This review documents the emergence of P. ramorum based on evolutionary and population genetic analyses. Currently infection by P. ramorum occurs only in Europe and North America and three clonal lineages are distinguished: EU1, NA1 and NA2. Ancient divergence of these lineages supports a scenario in which P. ramorum originated from reproductively isolated populations and underwent at least four global migration events. This recent work sheds new light on mechanisms of emergence of exotic pathogens and provides crucial insights into migration pathways.

Prospero, S., Grunwald, N. J., Winton, L. M., and Hansen, E. M. 2009. Migration patterns of the emerging plant pathogen Phytophthora ramorum on the West Coast of the United States of America. Phytopathology 99:739-749. Phytophthora ramorum (oomycetes) is the causal agent of sudden oak death and ramorum blight on trees, shrubs, and woody ornamentals in the forests of coastal California and southwestern Oregon and in nurseries of California, Oregon, and Washington. In this study, we investigated the genetic structure of P. ramorum on the West Coast of the United States, focusing particularly on population differentiation potentially indicative of gene flow. In total, 576 isolates recovered from 2001 to 2005 were genotyped at 10 microsatellite loci. Our analyses of genetic diversity and inferences of reproductive mode confirm previous results for the Oregon and California populations, with the strong majority of the genotypes belonging to the NA1 clonal lineage and showing no evidence for sexual reproduction. The high incidence of genotypes shared among populations and the lack of genetic structure among populations show that important large-scale, interpopulation genetic exchanges have occurred. This emphasizes the importance of human activity in shaping the current structure of the P. ramorum population on the West Coast of the United States.

International audience; The Landes de Gascogne forest (southwestern France) is the largest maritime pine (Pinus pinaster) plantation in Europe. Armillaria root disease (Armillaria ostoyae) has been reported since the early 1920s in the coastal area (western sector), but its incidence over the last 20 years has increased in the eastern sector. We investigated the genetic structure of the A. ostoyae population in this forest, focusing particularly on geographical differentiation potentially indicative of disease expansion in this area. In total, 531 isolates obtained from mycelial fans on symptomatic trees or undecayed stumps in 31 different disease foci were genotyped at five microsatellite loci. In 20 of these disease foci, a single genotype dominated, reflecting a predominantly clonal local spread of A. ostoyae. By contrast, at the regional scale, A. ostoyae probably spreads mostly via basidiospores (sexual spores), as no genotype common to several disease foci was identified. The absence of a clear pattern of isolation by distance may indicate either substantial gene flow or stochastic colonisation independent of spatial distance. The gradient of genetic diversity from the coast inwards and the greater genetic divergence of the eastern disease foci are consistent with the expansion of the A. ostoyae population from the coast eastwards.