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Populus nigra is an important autochthonous woody plant that can be grown as a renewable energy source. The possibility of its improvement through intraspecific hybridization was tested. Differences in biomass production, growth parameters, Melampsora larici-populina rust resistance and drought tolerance were evaluated among 19 intraspecific hybrids from controlled crosses, 2 clones selected from natural populations and the “MAX 4” clone (P. nigra × P. maximowiczii). These P. nigra clones from controlled crosses were chosen from more than 2000 hybrid individuals whose parents were selected from natural populations in the Czech Republic. A field trial was set up in Pruhonice, Czech Republic (320 m a.s.l., 591 mm rainfall annually, mean annual temperature of 9.5 °C). The planting density was 6061 plants ha-1, and the plants were coppiced three times at 3-year intervals. The trial was irrigated only during its establishment. Among the clones, significant differences were found in all the evaluated traits over three rotations. An average dry matter yield of the best clone “MAX 4” was 12.8 t ha-1 yr-1 over three harvests. The best black poplar clone reached up to 9.4 t ha-1 yr-1 in three harvests. Rust resistance was constant over 9 years and high for the three P. nigra clones (two from controlled crosses and one from natural populations). Moreover, the best P. nigra clones from controlled crosses showed higher drought tolerance than the “MAX 4” clone. Breeding progress was confirmed, and most of the P. nigra clones from controlled crosses performed better than the clones selected from natural populations. The trial validated the suitability of natural populations for use as gene sources for intraspecific hybridization and as sources of clones with traits comparable with those of interspecific clones. These new P. nigra clones can replace allochthonous clones in areas where autochthonous P. nigra populations are threatened by introgression.

Melampsora larici-populina is a macrocyclic rust, and the haploid stage with two nuclei and the diploid of mononuclear sequentially occur annually. During the preservation of dry urediniospores at −80°C, we found that one isolate, ΔTs06, was different from the usual wild-type isolate Ts06 at −20°C because it has mixed polykaryotic urediniospores. However, the other spores, including the 0, I, III, and IV stages of a life cycle, were the same as Ts06. After five generations of successive inoculation and harvest of urediniospores from the compatible host Populus purdomii, the isolate ΔTs06 steadily maintained more than 20% multiple nucleus spores. To test the pathogenesis variation of ΔTs06, an assay of host poplars was applied to evaluate the differences between ΔTs06 and Ts06. After ΔTs06 and Ts06 inoculation, leaves of P. purdomii were used to detect the expression of small secreted proteins (SSPs) and fungal biomasses using quantitative real-time PCR (qRT-PCR) and trypan blue staining. ΔTs06 displayed stronger expression of five SSPs and had a shorter latent period, a higher density of uredinia, and higher DNA mass. A transcriptomic comparison between ΔTs06 and Ts06 revealed that 3,224 were differentially expressed genes (DEGs), 55 of which were related to reactive oxygen species metabolism, the Mitogen-activated protein kinase (MAPK) signaling pathway, and the meiosis pathway. Ten genes in the mitotic and meiotic pathways and another two genes associated with the “response to DNA damage stimulus” all had an upward expression, which were detected by qRT-PCR in ΔTs06 during cryopreservation. Gas chromatography–mass spectrometry (GC-MS) confirmed that the amounts of hexadecanoic acid and octadecadienoic acid were much more in ΔTs06 than in Ts06. In addition, using spectrophotometry, hydrogen peroxide (H2O2) was also present in greater quantities in ΔTs06 compared with those found in Ts06. Increased fatty acids metabolism could prevent damage to urediniospores in super-low temperatures, but oxidant species that involved H2O2 may destroy tube proteins of mitosis and meiosis, which could cause abnormal nuclear division and lead to multinucleation, which has a different genotype. Therefore, the multinuclear isolate is different from the wild-type isolate in terms of phenotype and genotype; this multinucleation phenomenon in urediniospores improves the pathogenesis and environmental fitness of M. larici-populina.

Transcription factors (TFs) have been shown to play important roles in determining poplar susceptibility. In this study, the transcript profiles of five resistance-related TF groups at different time points were investigated to study the roles of TFs in the compatible interaction between &lsquo, Robusta&rsquo, (Populus nigra &times, P. deltoides) and the virulent E4 race of Melampsora larici-populina. The susceptibility test indicated that the parasitic process of E4 could be divided into two representative time periods: the infection phase and the production phase. Bioinformatics analysis showed that in these two phases, E4 infection induced a network of TFs in &lsquo, Although some TFs responded rapidly and positively, most TFs did not respond to E4, especially during the infection phase. The ethylene, jasmonic acid, and auxin pathways were downregulated, while a calcium-binding protein was upregulated. No other significantly changed phytohormone-related genes were found, which was consistent with the pathological process in the absence of an immune response, suggesting that the lack of response of most TFs during the infection phase of E4 is related to the susceptibility of &lsquo

Melampsora larici-populina causes serious poplar foliar diseases called rust worldwide. Salicylic acid (SA) and jasmonic acid (JA) are important phytohormones that are related to plant defence responses. To investigate the transcriptome profiles of SA- and JA-related genes involved in poplar rust interaction, two tolerant poplars and one intolerant poplar were selected for this study. Weighted gene coexpression network analysis (WGCNA) was applied to characterize the changes in the transcriptome profiles and contents of SA and JA after infection with the virulent E4 race of M. larici-populina. In response to infection with the E4 race of M. larici-populina, tolerant symptoms were correlated with the expression of genes related to SA and JA biosynthesis, the levels of SA and JA, and the expression of defence-related genes downstream of SA and JA. Tolerant poplars could promptly regulate the occurrence of defence responses by activating or inhibiting SA or JA pathways in a timely manner, including regulating the expression of genes related to programmed cell death, such as Kunitz-type trypsin inhibitor (KTI), to limit the growth of E4 and protect themselves. WGCNA suggested that KTI might be regulated by a Cytochrome P450 family (CYP) gene. Some CYPs should play an important role in both JA- and SA-related pathways. In contrast, in intolerant poplar, the inhibition of SA-related defence signalling through increasing JA levels in the early stage led to continued inhibition of a large number of plant–pathogen interaction-related and signalling-related genes, including NBS-LRRs, EDS1, NDR1, WRKYs, and PRs. Therefore, timely activation or inhibition of the SA or JA pathways is the key difference between tolerant and intolerant poplars.

The TIFY domain contains approximately 36 conserved amino acids that form the core motif TIF[F/Y]XG, and they were reported to play important roles in plant growth, tissue development and defence regulation. Moreover, more and more evidence has shown that some members of the TIFY gene family perform their functions by modulating plant hormone signalling pathways. Poplar trees are found worldwide, and they comprise approximately 30 species. Benefit from the importance of poplar and its advanced platform, this tree is considered to be the model perennial plant. Here, we conducted a genome-wide identification of TIFY genes in poplar, and 24 TIFY genes were found. These 24 TIFY genes were assigned to different subfamilies according to the presence or absence of domains and motifs that they harboured. Careful analyses of their locations, structures, evolution and duplication patterns revealed an overview of this gene family in poplar. The expression profiles of these 24 TIFY genes were then analysed in different tissues using publicly available expression data; their expression profiles following different JA/SA treatments and infection with leaf rust pathogen were also carefully examined by qRT-PCR assays. Based on their expression profiles, the functions of a number of TIFY genes could be predicted. By performing this study, we have provided valuable information for further functional characterisation of TIFY genes in poplar and candidate genes for the improvement of poplar disease resistance.

International audience; The rust fungi Melampsora larici-populina (Mlp) and Melampsora allii-populina (Map) are the main phytosanitary constraints for commercial poplar cultivation in Europe. Although Mlp is more aggressive and prevalent than Map, the two species may co-infect the same poplar tree or even the same poplar leaf, making the epidemiological surveys of each species difficult to achieve. In this study, a new duplex real-time PCR assay targeting each species was developed, based on single-copy genes. This test proved to be specific, inclusive and was successfully used to detect and quantify each species, starting from urediniospore samples or directly from infected poplar leaves, with or without visible symptoms. This new molecular tool was also assessed for comparative studies of time-course infection experiments on artificially inoculated poplar leaf discs. These studies showed that the growth dynamics of Map were significantly slower when the two species were co-inoculated on the same leaf disc, confirming that Map is less aggressive than Mlp.

Background MicroRNAs (miRNAs) are small non-coding RNAs that regulate the gene expression of target mRNAs involved in plant growth, development, and abiotic stress and pathogen responses. Previous studies have reported miRNAs in Populus that respond to abiotic stresses, such as cold, heat, drought, flooding, high salt and mechanical stress. However, little is known about the regulatory roles of these molecules in the Populus response to the stress of foliar rust fungal infection. Here, we identified the miRNA profiles of Populus after inoculation with Melampsora larici-populina using high-throughput sequencing and bioinformatics analysis. Quantitative real-time PCR (qRT-PCR) was used to validate the expression levels of 10 miRNAs. Results A total of 90 known miRNAs belonging to 42 families and 378 novel miRNAs were identified from three small-RNA libraries of Populus szechuanica infected with M. larici-populina isolates Sb052 and Th053 and a control. Comparative analysis revealed that the expression of 38 known miRNAs and 92 novel miRNAs in P. szechuanica after infection with different rust fungus isolates showed significant differences, and more miRNAs were suppressed during rust infection. Among the differentially expressed miRNAs, 7 known and 20 novel miRNAs were relevant to the rust fungus infection, and according to KEGG (Kyoto Encyclopaedia of Genes and Genomes) pathway analysis, these miRNAs primarily regulate genes encoding disease-resistance proteins, serine/threonine protein kinases, transcription factors, and related proteins. QRT-PCR analysis indicated that most miRNAs were up-regulated in the Sb052 library and down-regulated in the Th053 library at 48 h post-inoculation (hpi). Conclusions These results demonstrate that the expression of miRNAs was altered in poplar under stress associated with M. larici-populina infection, and different temporal dynamics were observed in incompatible and compatible libraries. These findings suggest important roles for miRNA regulation in Populus upon infection with foliar rust fungus. Electronic supplementary material The online version of this article (doi:10.1186/s12864-015-1891-8) contains supplementary material, which is available to authorized users.

Pinon, J., Frey, P., and Husson, C. 2006. Wettability of poplar leaves influences dew formation and infection by Melampsora larici-populina. Plant Dis. 90:177-184. The wettability of leaves of 60 poplar (Populus spp.) clones from sections Aigeiros (black) and Tacamahaca (balsam) and their hybrids was assessed by measuring the contact angle of calibrated water droplets on their abaxial leaf surface. The frequency and duration of dew on these clones were recorded in two nurseries. Black poplar leaves had the highest wettability, with relatively flat-shaped water droplets and a short drying time, in both the laboratory and nursery. Conversely, round-shaped water droplets on balsam poplars dried more slowly in the laboratory. In the nursery, dew was more frequent and persisted longer. First-generation intersectional hybrids behaved like balsam poplars, but successive backcrosses with black poplar (P. deltoides) resulted in clones with leaves that were more wettable and possessed less frequent and persistent dew. Infection by Melampsora larici-populina on two hybrid clones (‘Robusta’ and ‘Beaupre’), assessed by the number of uredinia in the laboratory, was dependent on the duration of leaf moisture. Maximum infection occurred when water was present on the leaves for 12 h. Leaf wettability should be considered as an additional trait when breeding poplar for durable resistance to M. larici-populina, as this characteristic is likely to be a useful defense against all pathotypes of the pathogen.

International audience; • In order to elucidate the genetic control of resistance to Melampsora larici-populina leaf rust in hybrid poplars, a Populus deltoides × P. trichocarpa F1 progeny was analysed for qualitative and quantitative rust resistances. • This progeny was evaluated for three components of quantitative resistance (latent period, uredinia number and uredinia size) to seven M. larici-populina strains in controlled conditions, and for one component of field susceptibility (rust colonization on the most infected leaf). • One qualitative resistance locus inherited from P. deltoides, R1, was localized on the genetic map. It segregates 1 : 1 in the F1 progeny and is effective against four of the studied strains. QTL analysis was performed separately on R1 and r1 genotype subsets. An additional detection was conducted on the entire F1 progeny for the three strains able to overcome R1 and for MAX2. A total of nine QTLs were detected. Two had large, broad-spectrum effects. One (RUS) is inherited from the P. trichocarpa parent; the other is inherited from P. deltoides and colocalized with R1. Seven QTLs had only limited and specific effects. Significant interaction effects were detected mainly between the two major QTLs. • Implications of these results for durable resistance breeding strategies, and possible benefits from the Populus genome sequence, are discussed.

1for correspondence) Summary The effects of poplar varietal mixture on infection levels caused by Melampsora larici-populina and on plant growth were studied during two consecutive growing seasons. The varietal mixture consisted of an intimate mixture of three poplar clones, Populus x interamericana ‘Boelare’, ‘Rap’ and Populus candicans ‘NNT’, expressing different race-specific resistances. Infection levels of M. larici-populina were assessed in monoclonal and polyclonal plots. Plant heights were determined at the end of each growing season. Infection levels in the growing season of the first year were not significantly different between monoclonal and polyclonal plots at the end of the epidemic. During the second growing season, a naturally occurring complex pathotype, able to infect clones ‘Boelare’ and ‘Rap’, invaded some plots prematurely and changed the composition of pathogen population. During this season, four infection assessments were conducted. Infection reduction in polyclonal plots compared with monoclonal plots was observed on ‘Boelare’ and P. candicans plants. Nevertheless, no significant difference was observed for growth between monoclonal and polyclonal plots. Biological factors which may be involved in the reduction of infection levels in polyclonal plots are dilution of susceptible individuals, physical barriers caused by resistant plants and cross-protection. A high auto-infection ratio may have partly hidden the positive effects of the clonal mixture.

Most rust fungi have a complex life cycle involving up to five different spore-producing stages. The telial stage that produces melanized overwintering teliospores is one of these and plays a fundamental role for generating genetic diversity as karyogamy and meiosis occur at that stage. Despite the importance of telia for the rust life cycle, almost nothing is known about the fungal genetic programs that are activated in this overwintering structure. In the present study, the transcriptome of telia produced by the poplar rust fungus Melampsora larici-populina has been investigated using whole genome exon oligoarrays and RT-qPCR. Comparative expression profiling at the telial and uredinial stages identifies genes specifically expressed or up-regulated in telia including osmotins/thaumatin-like proteins (TLPs) and aquaporins that may reflect specific adaptation to overwintering as well numerous lytic enzymes acting on plant cell wall, reflecting extensive cell wall remodeling at that stage. The temporal dynamics of karyogamy was followed using combined RT-qPCR and DAPI-staining approaches. This reveals that fusion of nuclei and induction of karyogamy-related genes occur simultaneously between the 25 and 39 days post inoculation time frame. Transcript profiling of conserved meiosis genes indicates a preferential induction right after karyogamy and corroborates that meiosis begins prior to overwintering and is interrupted in Meiosis I (prophase I, diplonema stage) until teliospore germination in early spring.

Fungal colonization can significantly affect the secondary metabolism of the host plants. We tested the impact of a common below-ground symbiosis, i.e., ectomycorrhiza formation, on poplar leaf chemical components that are involved in the defence against a common disease, i.e., rust fungi, in N-deficient soil. A rust-susceptible poplar clone (Populus trichocarpa x deltoides 'Beaupre') was (a) non-associated with ectomycorrhizal fungus (EM) Hebeloma mesophaeum (Pers.) Quelet MUN and non-infected with rust fungus Melampsora larici-populina Kleb. (isolate 98AG31), (b) associated with EM, (c) inoculated with rust fungus and (d) associated with EM and inoculated with rust fungus. Poplar leaves were analysed by photometric and mass spectrometric techniques (liquid chromatography-tandem mass spectrometry (LC-MS/MS), pyrolysis-field ionization mass spectrometry (Py-FIMS)). Both rust infection and mycorrhiza formation led to increased proportions of condensed tannins in relation to total phenolics (13% in the control, 18-19% in the fungal treatments). In contrast, salicylic acid concentration (6.8 mu g g(-1) in the control) was higher only in the rust treatments (17.9 and 25.4 mu g g(-1) with rust infection). The Py-FIMS analysis revealed that the rust-infected treatments were significantly separated from the non-rust-infected treatments on the basis of six flavonoids and one lipid. The relative abundance of these components, which have known functions in plant defence, was decreased after rust infection of non-mycorrhizal plants, but not in mycorrhizal plants. The results indicate that the ectomycorrhizal formation compensated the rust infection by a decrease in the flavonoid syntheses. The study provides new evidence for an interactive response of mycorrhizal colonization and infection with rust fungi in the metabolism of poplar.

International audience; Fungal plant pathogens, especially rust fungi (Pucciniales), are well known for their complex life cycles, which include phases of sexual and asexual reproduction. The effect of asexual multiplication on population genetic diversity has been investigated in the poplar rust fungus Melampsora larici-populina using a nested hierarchical sampling scheme. Four hierarchical levels were considered: leaf, twig, tree and site. Both cultivated and wild poplar stands were sampled at two time points at the start and end of rust epidemics. A total of 641 fungal isolates was analysed using nine microsatellite markers. This study revealed that the genetic signature of asexual multiplication in the wild poplar stand was seen only at lower hierarchical levels (leaf and twig). Moreover, we observed an erosion of clonal structure through time, with an increase in both gene and genotypic diversity. New genotypes contributed to host infection over time, which demonstrates the importance of allo-infection in the epidemic process in this host-pathogen system. Compared with the wild stands, the nearly lack of detection of clonal structure in the cultivated stands reflects the higher infection level on cultivated poplars. More generally, this genetic analysis illustrates the utility of population genetics approach for elucidating the proportion of asexual reproduction in the multiplication of isolates during an epidemic, and for proper quantification of asexual dispersal in plant pathogens.

Many factors can impact the genetic structures and population dynamics of fungal plant pathogens. Here we illustrated some of them at different spatial scales for the poplar rust fungus Melampsora larici-populina. At the scale of France, two main genetic groups were found. The first one infects only wild hosts and results from natural evolution of rust populations. The second one was formed after the R7 resistance breakdown, which is carried by the cultivar ?Beaupré?. This group exhibited a high proportion of virulent 7 individuals and presented signs of selection and demographic expansion; these signs indicate the recent invasion of individuals from this group on both wild and cultivated poplars in the northern half of France. At a regional scale, we focused on a contact zone between the ?cultivated? and the ?wild? genetic groups, located in the Alps. Upstream the Durance River valley, the influence of landscape has been highlighted by the effect of the Ecrins range which protects the ?wild? group located on the east side from the invasion of individuals from the ?cultivated? group, which arise from the northwest. Downstream the valley the annual epidemic wave was shown to be composed of admixed individuals from ?wild? and ?cultivated? groups, originating from both sides of the Ecrins range. Lastly we assessed the genetic consequences of the colonization wave. We evidenced a gradual increase of genetic differentiation with the epidemic source and a loss of genetic diversity. This work highlights the need of combining population genetics and epidemiology to characterize demographic processes and their genetic consequences, La structure génétique et la dynamique des populations des champignons phytopathogènes peuvent être influencées par de nombreux facteurs, ce que nous avons illustré à différentes échelles chez Melampsora larici-populina, l'agent de la rouille du peuplier. A l'échelle de la France, deux principaux groupes génétiques se différencient. Le premier, inféodé aux hôtes sauvages, est le produit de l'évolution des populations en conditions naturelles. Le second, formé suite au contournement de la résistance R7 portée par le cultivar "Beaupré", se caractérise par une forte proportion d"individus virulents 7 et présente des signatures de sélection et d'expansion démographique, témoins de l'invasion de ce groupe sur tous les peupliers (y compris sauvages) de la moitié nord de la France. A une échelle plus restreinte, nous avons examiné une zone de contact située dans les Alpes, où la délimitation entre les groupes « cultivé » et « sauvage » était la plus marquée. En testant l'effet du paysage, nous avons montré que le massif des Ecrins protégeait le groupe « sauvage » situé à l'est, en amont de la vallée de la Durance, de l'invasion des individus au profil « cultivé » venant du nord-ouest. Il n'en est pas de même dans la partie aval de cette vallée, colonisée annuellement par une vague épidémique, issue de ces deux groupes génétiques présents en amont de part et d'autre du Massif des Ecrins. En dernier lieu nous avons examiné les conséquences génétiques des évènements de colonisation, qui se traduisent par une augmentation de la différenciation par rapport à la source de l'épidémie ainsi qu'une érosion de la diversité génétique. Ce travail original a permis de souligner l'importance de combiner les approches d'épidémiologie et de génétique des populations pour caractériser au mieux les processus démographiques et leurs conséquences génétiques

La structure génétique et la dynamique des populations des champignons phytopathogènes peuvent être influencées par de nombreux facteurs, ce que nous avons illustré à différentes échelles chez Melampsora larici-populina, l'agent de la rouille du peuplier. A l'échelle de la France, deux principaux groupes génétiques se différencient. Le premier, inféodé aux hôtes sauvages, est le produit de l'évolution des populations en conditions naturelles. Le second, formé suite au contournement de la résistance R7 portée par le cultivar "Beaupré", se caractérise par une forte proportion d"individus virulents 7 et présente des signatures de sélection et d'expansion démographique, témoins de l'invasion de ce groupe sur tous les peupliers (y compris sauvages) de la moitié nord de la France. A une échelle plus restreinte, nous avons examiné une zone de contact située dans les Alpes, où la délimitation entre les groupes « cultivé » et « sauvage » était la plus marquée. En testant l'effet du paysage, nous avons montré que le massif des Ecrins protégeait le groupe « sauvage » situé à l'est, en amont de la vallée de la Durance, de l'invasion des individus au profil « cultivé » venant du nord-ouest. Il n'en est pas de même dans la partie aval de cette vallée, colonisée annuellement par une vague épidémique, issue de ces deux groupes génétiques présents en amont de part et d'autre du Massif des Ecrins. En dernier lieu nous avons examiné les conséquences génétiques des évènements de colonisation, qui se traduisent par une augmentation de la différenciation par rapport à la source de l'épidémie ainsi qu'une érosion de la diversité génétique. Ce travail original a permis de souligner l'importance de combiner les approches d'épidémiologie et de génétique des populations pour caractériser au mieux les processus démographiques et leurs conséquences génétiques

International audience; Forty-one microsatellite markers were developed for the poplar rust fungus Melampsora larici-populina. The genome sequence of M. larici-populina strain 98AG31 was screened in silico for microsatellite loci using Magellan software. Thirty-nine loci, evaluated on a sexual population of 35 M. larici-populina isolates sampled on larch trees in the French Alps, were identified to be highly polymorphic. These markers revealed two to 34 alleles, with an average of 7.23 alleles per locus. The estimated heterozygosity ranged from 0.029 to 0.958 across the 41 microsatellite loci. These markers will be highly useful for population genetics studies of M. larici-populina.

International audience; The course of the infection and development of the biotrophic fungus Melampsora larici-populina on leaf tissue from the hybrid poplar Populus deltoides × P. nigra ‘Ogy’ was monitored at the histological level. Leaf disks were inoculated with one of two rust physiological races (E1 and E2), resulting in interactions that were either incompatible (race E1) or compatible (race E2). In the compatible interaction, the fungus rapidly colonized the leaf without inducing any apparent host response. Symptoms appeared on the leaf several days after inoculation just prior to spore dissemination. The incompatible interaction was characterized by the early collapse and disorganization of cytoplasm of infected cells 17 h after inoculation and within 2 h after the appearance of the first haustoria. Resistance to M. larici-populina was mediated through a hypersensitive response, since it was extremely localized and involved only the few cells that were in the immediate vicinity of each infected cell.

International audience; Dispersal has a great impact on the genetic structure of populations, but remains difficult to estimate by direct measures. In particular, gradual and stochastic dispersal are often difficult to assess and to distinguish, although they have different evolutionary consequences. Plant pathogens, especially rust fungi, are suspected to display both dispersal modes, though on different spatial scales. In this study, we inferred dispersal capacities of the poplar rust fungus Melampsora larici-populina by examining the genetic diversity and structure of 13 populations from eight European and two overseas countries in the Northern hemisphere. M. larici-populina was sampled from both cultivated hybrid poplars and on the wild host, Populus nigra. The populations were analyzed with 11 microsatellite and 8 virulence markers. Although isolates displayed different virulence profiles according to the host plant, neutral markers revealed little population differentiation with respect to the type of host. This suggests an absence of reproductive isolation between populations sampled from cultivated and wild poplars. Conversely, studying the relationship between geographic and genetic structure allowed us to distinguish between isolation by distance (IBD) patterns and long distance dispersal (LDD) events. The European populations exhibited a significant IBD pattern, suggesting a regular and gradual dispersal of the pathogen over this spatial scale. Nonetheless, the genetic differentiation between these populations was low, suggesting an important gene flow on a continental scale. The two overseas populations from Iceland and Canada were shown to result from rare LDD events, and exhibited signatures of strong founder effects. Furthermore, the high genetic differentiation between both populations suggested that these two recent introductions were independent. This study illustrated how the proper use of population genetics methods can enable contrasted dispersal modes to be revealed.

International audience; Qualitative resistance to Melampsora larici-populina leaf rust inherited from North American species Populus deltoides did not allow for durable control of this pathogen in interspecific hybrid cultivars. Despite significant levels of strain-specificity, quantitative resistance would exert lower selection pressures on the pathogen populations, and hence could be more durable. Previous studies restricted to a large P. × interamericana (i.e., P. deltoides × Populus trichocarpa) F1 family revealed that the presence of R1, a segregating defeated qualitative resistance gene inherited from P. deltoides, had major beneficial effects on quantitative resistance. The present study was based on 14 F1 families from a 4 × 5 P. deltoides × P. trichocarpa factorial mating design where at least four defeated qualitative resistances segregate 1:1. Even though quantitative resistance assessments were conducted in the laboratory with a M. larici-populina strain able to overcome these qualitative resistances, their presence had a significant effect on the mean level and on the genetic variability for quantitative resistance. One unprecedented result is the identification of a defeated qualitative resistance which presence is associated with lower levels of quantitative resistance. Possible inferences on the nature of the genetic relationship between both resistance types are discussed.

We developed 15 microsatellite loci in the poplar rust fungus, Melampsora larici-populina , using two enrichment protocols. Polymorphism of each locus was assessed on a panel of 30 isolates, comprising three subpanels (world, regional and local scales). Thirteen loci were polymorphic with three to eight alleles detected. The 15 loci were also tested on five related Melampsora species, M. allii-populina , M. medusae f. sp. deltoidae , M. larici-tremulae , M. rostrupii and M. pinitorqua , and partial or global cross-amplification events were detected.

In September 2002, yellow spots were observed on the leaf surface of a hybrid poplar (Populus maximowiczii Henry × P. balsamifera L.) grown at the Berthier forest nursery (46°2′N, 73°11′W) in the St. Lawrence Valley (Lanaudière Region, Québec, Canada). Disease severity was low, but the pathogen was present on a hybrid that was previously thought to be resistant to Melampsora medusae Thuem, the only reported poplar rust in eastern North America. Uredinia typical of a Melampsora sp. were observed on the abaxial leaf surface. The observed urediniospores were longer (32 to 48 μm) than the expected range for M. medusae (23 to 35 μm) and possessed an apical bald spot; thick paraphyses were also observed. These characteristics are diagnostic of M. larici-populina Kleb (2). Samples were deposited in the National Mycological Herbarium of Canada (DAOM 232107 and 232108) and in the Quebec Forest Biology Herbarium (QFB14703 and 14704). DNA was extracted from uredinia, and the internal transcribed spacer (ITS) of the ribosomal RNA gene was amplified and sequenced (GenBank Accession Nos. AY429656 and AY429657). There was a 100% match between the two sequences obtained and that of M. larici-populina (GenBank Accession No. AY375267), but there was approximately 12% divergence with the ITS sequence of M. medusae (GenBank Accession No. AY375273-5). This is the first report of M. larici-populina in eastern North America. This fungus was reported on P. trichocarpa × P. deltoides hybrids in the western United States in the early 1990s (1). It appears that M. larici-populina can overwinter in Québec because it was observed again at the nursery in September 2003. The occurrence of M. larici-populina in eastern North America has direct implications for the poplar industry since the host specificities of M. medusae and M. larici-populina differ; P. balsamifera and P. maximowiczii are sensitive to M. larici-populina (3). Hybrids with P. balsamifera or P. trichocarpa components may be particularly at risk. References: (1) G. Newcombe and G. A. Chastagner. Plant Dis. 77:532, 1993. (2) J. Pinon. Eur. J. For. Pathol. 3:221, 1973. (3) J. Pinon. Silvae Genet. 41:25, 1992.

A plant's capability to develop ectomycorrhizal symbiosis is under the control of both genetic and environmental factors. In order to determine the roles played by these different factors, we have performed a quantitative genetic analysis of the ability of poplar trees to form ectomycorrhizas. Quantitative genetics were applied to an interspecific family of poplar for which the two parental genetic maps had already been described, and for which data analyses concerning fungal aggressors were obtained. Quantitative trait loci (QTL) related to ectomycorrhiza formation were identified and located in the genetic maps of the two parents. One QTL was located at a linkage group of the genetic map of Populus trichocarpa showing a high concentration of several QTL involved in the pathogenic interaction with the fungus Melampsora larici-populina, the causal agent of leaf rust.

Inoculated excised leaf disk technique allows decomposition of poplar partial resistance to Melampsora larici-populina leaf rust into key epidemiological components such as latent period (LP), uredinia number (UN), uredinia size (US), and spore production (SP) for a given M. larici-populina strain under controlled environmental conditions. Three hundred thirty-six genotypes from an interspecific Populus deltoides × P. trichocarpa F1 progeny segregating for complete resistance to M. larici-populina strain 93ID6 were inoculated with M. larici-populina strain 93CV1. This strain was able to infect the whole family, except few probable recombinants. LP, final UN, and final US after one infectious cycle proved to be relevant complementary descriptors of partial resistance. Area under the disease progress curve and other parameters of uredinia appearance dynamics did not yield additional information. Indirect assessment of SP by US scoring was reliable and easy to access compared with direct spore counting. UN was the only trait for which a doubling of the inoculum pressure level had a significant effect, leading to greater differentiation between genotypes. Consistent with previous studies is the clear relationship between presence of complete resistance against M. larici-populina strain 93ID6 and higher partial resistance to M. larici-populina strain 93CV1 (32% longer LP, 76% smaller UN, and 34% smaller US). In the subpopulation compatible with 93ID6, bimodal distribution of genotypic means for US suggested implication of a major gene inherited from the P. trichocarpa parent. Residual variation was noted for the three epidemiological components, suggesting that additional genes might condition these quantitative traits.

International audience; Partial resistance of Populus nigra L. to three races of the foliar rust Melampsora larici-populina Kleb. was studied in a field trial and in laboratory tests, using a collection of P. nigra originating from different places throughout France. No total resistance was found. The partial resistance was split into epidemiological components, which proved to be under genetic control. Various patterns of association of epidemiological components values were found. Principal components analysis revealed their relationships. Only 24% of the variance of the field susceptibility could be explained by the variation of the epidemiological components of susceptibility. This variable was significantly correlated with susceptibility to the most ancient and widespread race of the pathogen, and with the variables related to the size of the lesions of the different races. Analysis of variance showed significant differences in susceptibility between regions and between stands within one region. Up to 20% of variation was between regions, and up to 22% between stands, so that these genetic factors appeared to be more differentiated than the neutral diversity (up to 3.5%; Legionnet & Lefevre, 1996). However, no clear pattern of geographical distribution of diversity was detected.

Not available, Les protéines de réserve végétatives (VSPS) majeures de populus x euramericana ont été mises en évidence dans l'écorce, le bois, les bourgeons et les racines. Si dans la tige, les VSPS de p. euramericana montrent peu de spécificité tissulaire, la vacuole constitue leur seul site de stockage. Les protéines de réserve végétatives de l'écorce sont des protéines solubles, majeures durant la phase hivernale, et composées d'un seul type de sous-unités de 32, 34, 36 et 38 kda. Elles constituent probablement les glycoformes d'un polypeptide de 29 kda portant des chaines oligosaccharidiques de nature, de nombre, ou de longueur variable. D'autre part, nous avons montré qu'il existe aussi des homologies structurales et immunologiques entre les VSPS du bois et de l'écorce de p. euramericana. La biosynthèse des VSPS de p. euramericana apparait strictement régulée. Elle intervient lors du changement de photopériode en jours courts, et durant l'hydrolyse massive des protéines foliaires. Toutefois, l'accumulation des VSPS se déroule aussi lorsque les relations source-puits sont affectées par une attaque parasitaire des feuilles par melampsora larici-populina

Les populations raciales de L larici-populina ont ete etudiees pendant 4 ans (pres de 7000 identifications), dans plusieurs pepinieres de l'est de la France. La race E1 est ubiquiste en France (et tres probablement en Europe occidentale), la race E2 existe au moins dans la moitie nord de la France, en Belgique, aux Pays-Bas, et la race E3 semble presente dans l'ouest et le sud-ouest de la France et en Italie, dans la vallee du Po. Les races E2 et E3 ont une frequence irreguliere d'une annee a l'autre sur le meleze, l'hote alternant (tableau I). Il existe un certain lien entre les populations raciales du meleze et celles des peupliers (tableau I et II) (...)

Not available, Les protéines de réserve végétatives (VSPS) majeures de populus x euramericana ont été mises en évidence dans l'écorce, le bois, les bourgeons et les racines. Si dans la tige, les VSPS de p. euramericana montrent peu de spécificité tissulaire, la vacuole constitue leur seul site de stockage. Les protéines de réserve végétatives de l'écorce sont des protéines solubles, majeures durant la phase hivernale, et composées d'un seul type de sous-unités de 32, 34, 36 et 38 kda. Elles constituent probablement les glycoformes d'un polypeptide de 29 kda portant des chaines oligosaccharidiques de nature, de nombre, ou de longueur variable. D'autre part, nous avons montré qu'il existe aussi des homologies structurales et immunologiques entre les VSPS du bois et de l'écorce de p. euramericana. La biosynthèse des VSPS de p. euramericana apparait strictement régulée. Elle intervient lors du changement de photopériode en jours courts, et durant l'hydrolyse massive des protéines foliaires. Toutefois, l'accumulation des VSPS se déroule aussi lorsque les relations source-puits sont affectées par une attaque parasitaire des feuilles par melampsora larici-populina

Ecophysiology of the uredial states of Melampsora larici-populina Kleb. and M. allii-populina Kleb. Reactions of M. larici-populina and M. allii-populina urediospores for germination and germ tubes growth to humidity, light and temperature are compared. Differences between the two species suggest that M. allii-populina is more thermophilic.