Biscogniauxia nummularia (Bull.) Kuntze is a fungus which induces strip-cankers on beech, commonly referred to as charcoal canker. The symptoms of infection are visible on the host tree’s bark as elongated, blackish bark lesions on the trunk and branches. Recent years have shown that, due to climate change causing local epidemics, the species is increasing its economic impact in Mediterranean regions. Until recently, B. nummularia was considered rare and uncommon in central Europe. However, in the last few years it has been noticed more often, mostly in coniferous trees, which are out of B. nummularia’s host range. A similar situation has been observed with the closely related species Biscogniauxia mediterranea (De Not.) Kuntze, which prior to 2017 had not been observed in central Europe at all. This study shows the genetic diversity of mid-European strains of Biscogniauxia spp. (based on the ITS, TEF1, TUB2 and ACT regions) and, as the first in Europe, presents a molecular investigation of this species isolated from coniferous trees. It is also the first attempt at estimating the potential impact of this pathogenic fungus on European forestry management in the close future.

The internal transcribed spacer (ITS) region is regarded as a formal fungal primary barcode with a high probability of the correct identification for a broad group of fungi. ITS sequences have been widely used to determine many fungal species and analysis of rDNA ITS is still one of the most popular tools used in mycology. However, this region is not equally variable in all groups of fungi; therefore, identification may be problematic and result in ambiguous data, especially in some species-rich genera of Ascomycota. For these reasons, identification based on rDNA ITS is usually complemented by morphological observations and analysis of additional genes. Reliable species identification of Ascomycota members is essential in diagnosing plant diseases, verifying air quality and the effectiveness of agronomic practices, or analyzing relationships between microorganisms. Therefore, the present study aimed to verify, using specific examples, the extent to which ITS sequence analysis is useful in species identification of pathogens and saprobionts from Ascomycota and demonstrate problems related to such identification in practice. We analyzed 105 ITS sequences of isolates originating from air and plant material. Basic local alignment search tool (BLASTn) significantly contributed to the reliable species identification of nearly 80% of isolates such as <em>Arthrinium arundinis</em>, <em>Beauveria bassiana</em>, <em>Boeremia exigua</em>, <em>Cladosporium cladosporioides</em>, <em>Epicoccum nigrum</em>, <em>Nigrospora oryzae</em>, <em>Sclerotinia sclerotiorum</em>, or <em>Sordaria fimicola </em>and members of the genera <em>Alternaria </em>and <em>Trichoderma</em>. However, for most isolates, additional morphological observations, information regarding the isolate origin and, where possible, a PCR with species-specific primers were helpful and complementary. Using our practical approach, we determined that ITS-based species identification and comparative analysis with GenBank sequences significantly helps identifying Ascomycota members. However, in many cases, this should be regarded as suggestive of a taxon because the data usually require the use of additional tools to verify the results of such analysis.

Box elder (Acer negundo) is a tree native to North America. In Europe it is considered a dangerous invasive species, and assigned to the highest (4th) category of environmental hazard (Tokarska-Guzik et al. 2012). The tree can threaten a wide range of ecosystems and compete with the native flora. The shoot dieback was observed on 20% of boxelder in July 2018 and 2019 in Bryzgiel (N53°59.963' E23°04.324') in NE Poland (Europe). Young trees (10-15 yr. old) with visible symptoms were observed in a small group on the rural roadside. Infected shoots were chlorotic. There were visible shallow cracks on the bark and brown discoloration in sapwood inside infected branches. Symptomatic shoots were collected in sterile envelopes, surface disinfected with 95% ethanol. Twelve fragments of wood were cut from the border of living and dead tissue, and then divided into 3-5 mm pieces, placed on PDA medium and incubated at 21°C. After 10 days ten Fusarium spp. strains were obtained. Pure cultures were derived by monosporic isolation. The identification of the isolates was initially based on morphology and molecular genotyping (Leslie & Summerell 2006). On PDA, strains produced white, dense, floccose aerial mycelium with a pink surface. The underside of the petri dish was brown. Growth of the colony was relatively slow and reached Ø 3.5 cm after two weeks. Microscopic observation revealed the presence of macroconidia located in a few orange sporodochia. Macroconidia were slightly curved, with dimensions of 38-45 µm × 3.2-3,5 µm, 4-5 septate, with well-formed foot cell and beak on the apex. On aerial hyphae, single intercalary chlamydospores were present. Microconidia were not found. Morphological identification was confirmed by sequencing the ITS regions, the TEF-1α and β-tubulin genes for representative isolates. Mycelia were grown on PDB and freeze dried prior to genomic DNA extraction using the CTAB method. Sequences of two isolates were deposited in GenBank as MN186748 and MN588156 for ITS; MZ191070 and MZ191072 for TEF-1α; and MZ191069 and MZ191071 for TEF-1α. BLASTn search in the NCBI database revealed 100, 98 and >99% similarities of ITS, TEF-1α and β-tubulin with F. lateritium isolates LC171689, KT350607 and FN554618 respectively. A pathogenicity test was conducted on five first year Ø 0.6-0.8cm shoots from a 10-year-old tree. Before inoculation their surface was disinfected with 95% ethanol. Then, bark of the twigs was split longitudinally with a sterile blade and pieces of 10-day-old aerial mycelium grown on PDA were applied on the wound sites. Control samples were inoculated with sterile distilled water only. Inoculated areas were covered with parafilm. First sign of infection was observed after three weeks, as a dark lesion in the place of inoculation and chlorosis. Three weeks later the brown ring on the sapwood was marked in the shoot cross-section. Morphologically identical to the original, F. lateritium isolate was reisolated from the infected tissues, thus fulfilling Koch’s postulates. F. lateritium is a species closely associated with trees and shrubs (Leslie & Summerell 2006). However, it has not been recorded on boxelder and this is the first report of F. lateritium causing dieback of boxelder maple. According to the Enemy Release Hypothesis (Elton 1958), new pathogens appearing on alien species can be an indicator of developing environmental resistance to the outlander, which indicate the grade of their domestication. This kind of notification poses a crucial role in invasion monitoring and the search for new biocontrol methods of invasive plant species.

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The aim of the research was to check whether the healing of bark-stripping wounds of the silver fir tree trunks reduces the share of wood-decomposing fungi, which may be the result of inter-species interactions. The study carried out in Gorce National Park in Polish Western Carpathians analyzed drill holes of sapwood from three types of wounds (fresh, healed and old) on fir trunks with a diameter at breast height (DBH) of 4.0–16.9 cm as a result of bark-stripping by red deer (Cervus elaphus). In the wood of fresh wounds Alternaria alternata (Fr.) Keissl. and Arthrinium arundinis (Corda) Dyko & B. Sutton had the largest share in mycobiota. Phompsis spp. and the species Sydowia polyspora (Bref. & Tavel) E. Müll. and Epicoccum nigrum Link were also isolated. The dominants in old wounds were Eutypa spp., Phomopsis spp. and Cylindrobasidium evolvens (Fr.) Jülich. Healed wounds were dominated by Trichoderma atroviride P. Karst, a fungus antagonistic to many fungal pathogens. Such properties are shared by A. arundinis, especially common in fresh wound wood. It seems that these fungi support the process of wounded tree regeneration (healing of wounds) and limit the activity of wood-decaying fungi in old age, which makes fir survival very high. Thus, even a strong red deer pressure cannot be considered the basic factor determining the dynamics of fir in this part of the Carpathians.