Search

The symptoms of ash dieback caused by the fungus Hymenoscyphus fraxineus include wilting of the foliage followed by dieback of shoots, twigs and branches. Necroses in shoots are assumed to develop after infection through leaf petioles; however, clear evidence of this infection pathway has not yet been provided. Considering the multiple pathogen genotypes in dead ash petioles, we aimed to obtain a spatial overview of all H. fraxineus genotypes colonizing individual shoots and their corresponding petioles before leaf shedding to acquire precise information about the infection biology of H. fraxineus and its ability to cross the petiole-shoot junction. Individual genotypes of H. fraxineus were characterized by the analysis of microsatellites using DNA extracted directly from petiole segments or cultures isolated from the segments. We detected 150 different multilocus genotypes in 10 analysed shoots and their respective petioles; the highest number of genotypes was eight for a single petiole and three for a single shoot. The genotypes of most shoot lesions were identical to particular genotypes from the proximal segments of petioles, implicating the main pathway of shoot infections. To test whether the amount of colonized substrate or intraspecific competition have an effect on successful infection, genotypes that reached the most proximal end of the petioles were scored for the number of invaded petiole segments and for the number of other H. fraxineus genotypes co-occurring in the segments. However, the extent of colonization of the scored genotypes and intraspecific competition with other H. fraxineus strains did not influence pathogen success in entering the shoot. This study confirms that the majority of ash shoot infections are caused by genotypes of H. fraxineus originating from petioles. Compared to petioles, the frequency of shoot colonization as well as number of H. fraxineus genotypes in shoots was much lower. [ABSTRACT FROM AUTHOR]

Twenty-six commercial formulations of fungicides at six concentrations were evaluated in vitro for their efficacy on mycelial growth of Hymenoscyphus fraxineus (anamorph Chalara fraxinea). The results are presented as EC50, EC90 and minimal inhibitory concentration values; the comparisons with the recommended application concentrations showed that 10 of the 26 fungicides were highly effective in their ability to inhibit the mycelial growth of the pathogen. The eight most effective fungicides identified based on multiple comparisons analysis were azoxystrobin, bitertanol, captan (in two-component preparation with trifloxystrobin), difenoconazole, kresoxim-methyl, spiroxamine (in multicomponent preparation with tebuconazole and triadimenol), tebuconazole and trifloxystrobin. Azoxystrobin, difenoconazole, kresoxim-methyl, mancozeb, myclobutanil, pyrimethanil, tebuconazole and trifloxystrobin were selected to verify their effectiveness in in planta tests. During two field tests in nurseries, it was found out that tebuconazole (triazole), trifloxystrobin and kresoxim-methyl (strobilurins) and mancozeb (dithiocarbamate) were significantly effective against H. fraxineus. The best results were gained with tebuconazole, when mean percentage of diseased saplings was 16.3%, whereas in the control, it was 63.6%. The combination and alternation of fungicides from the triazole, strobilurine and dithiocarbamate chemical groups should be an effective tool for protecting ash saplings in forest nurseries. Moreover, exact timing of the treatment reflecting rainfall and development of first symptoms in the foliage will be also important. [ABSTRACT FROM AUTHOR]

Due to its high tolerance to acid rain, the North American Colorado blue spruce ( Picea pungens) has been among the most frequent substitute tree species used in reforestation programmes in spruce mountain forests in the Ore and Sudeten Mountains in the Czech Republic after destruction by sulphur dioxide pollution in the second half of the last century. This species was planted on more than 8800 ha of destroyed mountain forest stands in the Ore Mountains alone. New plantations prospered until a massive outbreak of the gemmamyces bud blight, first identified in 2009. The causal agent, Gemmamyces piceae, induced massive bud blight in infected trees - bud loss progressively increased to 70-80% or more, and because the trees were not able to replace the natural loss of older needles, they quickly withered and died. Many plantations disrupted by the pathogen had to be cut down. Although several resistant genotypes have been identified, the cultivation of this species in forest stands has no future in this area. Because this disease was also identified in several ornamental plantations in the area, the bud blight epidemic haunts public green space administrators and nursery and Christmas tree plantation owners. Other pathogens and pests, such as Lophodermium piceae, Sirococcus conigenus and Elatobium abietinum, have also been identified as involved in the damage to P. pungens. [ABSTRACT FROM AUTHOR]

The impact of ash dieback caused by Hymenoscyphus fraxineus on 17 provenances of Fraxinus excelsior and one provenance of Fraxinus angustifolia was studied in an extensive field trial established in the Czech Republic prior to the H. fraxineus invasion in 1999. A difference in the level of resistance to ash dieback between the species was found: F. angustifolia was significantly less affected by the disease than F. excelsior. Moreover, particular provenances of F. excelsior showed important differences in the level of resistance to H. fraxineus. A relationship between the impact of ash dieback and altitude was also discovered - the provenances from altitudes above 600 m a.s.l. were less affected by the pathogen than were the provenances from lower areas. No difference in the impact of the disease among provenances of F. excelsior from different ecotopes (ravine, calcareous ravine and alluvial) was found. Substantial among-tree variability in resistance to H. fraxineus was observed throughout the trial - promising genotypes (with crown defoliation up to 5%) were identified in all 18 tested provenances. In regard to this finding, it appears that the main source of resistance to the pathogen is probably at the individual genotype level in the trial. A secondary but massive attack by Hylesinus fraxini was identified in the trees that had been greatly damaged by ash dieback, and the beetle caused their health to deteriorate significantly. A significant negative effect of the presence of collar necroses caused by H. fraxineus and browse damage was also identified. [ABSTRACT FROM AUTHOR]

The population structure of Hymenoscyphus fraxineus, the causal agent of ash dieback, was assessed at four closely located sites in the Czech Republic. To analyse the genetic variation, one Swiss and one Norwegian population with known population structures were selected as reference points. The analysis was performed using 16 previously published and five newly developed microsatellite markers. The quality of the new markers was assessed by sequencing the flanking region, identifying the type of the mutation and analysing the inheritance and linkage between all pairs of loci. In addition, markers were tested on 45 Japanese strains to confirm their usability on native H. fraxineus populations in Asia. One of the new markers was monomorphic in all European populations and one marker exhibited a high percentage of null alleles in the Japanese samples. Twenty markers in the four Czech populations showed lower average gene diversity than in the other two European populations. One Czech population significantly differed from all the others, with a pairwise GST of approximately 0.2, the lowest allelic richness and very low average gene diversity, likely resulting from a founder effect. The amount of genetic differentiation between the four neighbouring Czech populations exceeds the expected value from previous Europe-wide population studies and suggests that local population dynamics can affect the population structure of H. fraxineus. [ABSTRACT FROM AUTHOR]

Chemical communication is mediated by sex-biased signals abundantly present in the urine, saliva and tears. Because most studies concentrated on the urinary signals, we aimed to determine the saliva proteome in wild Mus musculus musculus, to extend the knowledge on potential roles of saliva in chemical communication. We performed the gel-free quantitative LC-MS/MS analyses of saliva and identified 633 proteins with 134 (21%) of them being sexually dimorphic. They include proteins that protect and transport volatile organic compounds in their beta barrel including LCN lipocalins, major urinary proteins (MUPs), and odorant binding proteins (OBPs). To our surprise, the saliva proteome contains one MUP that is female biased (MUP8) and the two protein pheromones MUP20 (or 'Darcin') and ESP1 in individuals of both sex. Thus, contrary to previous assumptions, our findings reveal that these proteins cannot function as male-unique signals. Our study also demonstrates that many olfactory proteins (e.g. LCNs, and OBPs) are not expressed by submandibular glands but are produced elsewhere-in nasal and lacrimal tissues, and potentially also in other oro-facial glands. We have also detected abundant proteins that are involved in wound healing, immune and non-immune responses to pathogens, thus corroborating that saliva has important protective roles.