Eutypella parasitica R.W. Davidson and R.C. Lorenz is the causative agent of Eutypella canker of maple, a destructive disease of maples in Europe and North America. The fungus E. parasitica infects the trunk through a branch stub or bark wound. Because the fungal community may have an impact on infection and colonization by E. parasitica, the composition of fungi colonizing wood of dead branches of sycamore maple (Acer pseudoplatanus L.) was investigated in five sampling sites in Slovenia. Forty samples from each sampling site were collected between the November 2017 and March 2018 period. Isolations were made from the wood in the outer part of dead branches and from discoloured wood in the trunk that originated from a dead branch. Pure cultures were divided into morphotypes, and one representative culture per morphotype was selected for further molecular identification. From a total of 2700 cultured subsamples, 1744 fungal cultures were obtained, which were grouped into 212 morphotypes. The investigated samples were colonized by a broad spectrum of fungi. The most frequently isolated species were Eutypa maura (Fr.) Sacc., Eutypa sp. Tul. and C. Tul., Fusarium avenaceum (Fr.) Sacc., Neocucurbitaria acerina Wanas., Camporesi, E.B.G. Jones and K.D. Hyde and E. parasitica. In this study, we distinguished species diversity and the fungal community. There were no significant differences in the diversity of fungal species between the five sampling sites, and branch thickness did not prove to be a statistically significant factor in fungal species diversity. Nevertheless, relatively low Jaccard similarity index values suggested possible differences in the fungal communities from different sampling sites. This was confirmed by an analysis of similarities, which showed that the isolated fungal community distinctly differed between the five sampling sites and between the different isolation sources. Eutypella parasitica was isolated from all five investigated sampling sites, although Eutypella cankers were observed in only three sampling sites, indicating the possibility of asymptomatic infection.

Reports of sooty bark disease of maples caused by the fungus Cryptostroma corticale have recently been emerging from across Europe. The aims of our study were to describe the first report of sooty bark disease in Slovenia, to determine the pathogenicity of C. corticale, to confirm the optimum temperature for the growth of the fungus, and to determine the mass loss of Acer pseudoplatanus wood inoculated by C. corticale. We confirmed the presence of C. corticale on A. pseudoplatanus via morphological and molecular analysis. The optimal growth of C. corticale was measured in vitro on potato dextrose agar and was determined to occur at 25 °C. Pathogenicity tests were performed on 30 saplings of A. pseudoplatanus under two treatments, humid and drought stress, and the fungus was pathogenic in both treatments. The mean length of bark lesions and wood discoloration of the drought-stressed saplings was significantly greater than that in the humid treatment. Re-isolations of C. corticale were successful from all inoculated saplings, and thus Koch’s postulates were confirmed. The mass loss of A. pseudoplatanus wood was determined by mini-block test in a period of 10 weeks and was observed as minimal. Based on the results, we conclude that C. corticale is a weak and opportunistic pathogen that most likely expresses itself intensively under hot and dry conditions.

The fungus Pseudodidymella fagi is spreading in Europe and causing leaf blotch of European beech, Fagus sylvatica. Between 2008 and 2017, outbreaks of P. fagi were observed on European beech in Switzerland, Germany (also on F. orientalis), Austria, and Slovakia. In Slovenia, leaf blotch symptoms were first observed on F. sylvatica in 2018. P. fagi was identified as the causal agent of the observed symptoms in Slovenia by morphological examinations together with sequencing of the internal transcribed spacer (ITS) region of the rDNA. This study links the fungus to the expansion of the known distribution of the disease to Slovenia, and based on in vitro pathogenicity trials, also to a new potential host, Quercus petraea. The pathogenicity tests confirmed F. sylvatica and F. orientalis as hosts for P. fagi, but not Castanea sativa, where pathogenicity to F. orientalis was proved for first time in vitro. Although Koch’s postulates could not be proven for C. sativa, it seems to be partially susceptible in vitro because some of the inoculation points developed lesions. Additionally, damage to Carpinus betulus related to P. fagi near heavily infected beech trees was observed in vivo but was not tested in laboratory trials. Based on the results and our observations in the field, it is likely that P. fagi has a wider host range than previously thought and that we might be witnessing host switching.

The ten most frequently isolated fungi from the wood of the dead branches of Acer pseudoplatanus L. were tested in dual cultures to evaluate their in vitro antagonistic activity against Eutypella parasitica R.W. Davidson and R.C. Lorenz, the causative agent of a destructive disease of maples in Europe and North America. The tested fungi, treated also as challenge isolates, were Diaporthe sp., Eutypa sp., Eu. maura, E. parasitica, Fusarium avenaceum, Neocucurbitaria acerina, Neonectria sp., Peniophora incarnata, Petrakia irregularis, and Phomopsis pustulata. The antagonistic ability of each challenge isolate was evaluated by calculating an index of antagonism (AI) based on the interaction type in the dual cultures. The results of competition between the fungal isolates were quantified after re-isolations from the interaction zone (s). The dual cultures revealed two main types of competitive interactions: Deadlock, consisting of mutual inhibition after mycelial contact or at a distance, and replacement, reflecting in the inhibition of E. parasitica, followed by partial overgrowth by the replacing fungus. Statistical analysis showed significant differences in average AI and s of challenge isolates between different dual culture assays. Based on the results of the antagonism index, Eutypa sp., Eu. maura, Neonectria sp., and P. incarnata had the highest inhibitory effect on E. parasitica growth and were recognized as the most promising candidates for further biocontrol studies of E. parasitica. The mycelium of E. parasitica at the interaction zones remained mostly viable, except in dual cultures with Eutypa sp., F. avenaceum, and Neonectria sp., where re-isolations did not yield any colony of the E. parasitica isolate. Based on the results, we assume that E. parasitica is a weak competitor, which invests less energy in direct mycelial competition. We discuss the potential of the observed antagonists as a possible biocontrol of Eutypella canker of maple. Nevertheless, additional experiments should be performed for a solid conclusion about competitive ability of E. parasitica and usefulness of antagonists as biocontrol.