Piotr Bilański

Association of the pine-infesting Pissodes species with ophiostomatoid fungi in Poland

Bark weevils are consistently associated with various fungi. They act as effective vectors for root-rot fungi, the rust pathogen and the ophiostomatoid fungi. In comparison with bark beetles, the interaction between ophiostomatoid fungi and bark weevils has been poorly studied in Europe. This study aims to clarify the ties among ophiostomatoid fungi and their weevil vectors in Pinus sylvestris. Samples associated with three bark weevils, including Pissodes castaneus, P. piniphilus and P. pini were collected from seven pine stands in Poland. Fungi were isolated from laboratory-reared and field-collected beetles, larvae and galleries of weevils. Isolates were identified based on morphology, DNA sequence comparison for two gene regions (ITS, ß-tubulin) and phylogenetic analyses. Fourteen morphological and phylogenetic ophiostomatoid species were identified among the 1,219 isolates. These 14 species included 11 novel associations between fungi and bark weevils. The most commonly encountered fungal associates of bark Pissodes species were Leptographium procerum, Ophiostoma quercus and O. minus. The spectrum of fungal associates was similar in three bark weevil species, despite some differences between species. Except for P. castaneus, all species of bark weevil were frequently associated with ophiostomatoid fungi. Pissodes castaneus does appear to be a consistent vector of L. procerum and Sporothrix inflata while O. minus, O. quercus and O. cf. rectangulosporium were often found in association with P. piniphilus and P. pini.

Virulence of Hymenoscyphus albidus and H. fraxineus on Fraxinus excelsior and F. pennsylvanica.

European ash (Fraxinus excelsior) is currently battling an onslaught of ash dieback, a disease emerging in the greater part of its native area, brought about by the introduction of the ascomycete Hymenoscyphus fraxineus (= Hymenoscyphus pseudoalbidus). The closely-related fungus Hymenoscyphus albidus, which is indigenous to Europe, is non-pathogenic when in contact with F. excelsior, but could pose a potential risk to exotic Fraxinus species. The North American green ash (Fraxinus pennsylvanica) is planted widely throughout Europe and regenerates naturally within this environment but little is known about the susceptibility of this species to ash dieback. We performed wound inoculations with both fungi (nine strains of H. fraxineus and three strains of H. albidus) on rachises and stems of F. excelsior and F. pennsylvanica under field conditions in Southern Poland. Necrosis formation was evaluated after two months on the rachises and after 12 months on the stems. After inoculation of H. albidus, only small lesions (of up to 1.3 cm in length) developed on the F. excelsior and F. pennsylvanica rachises, but with no significant distinction from the controls. Hymenoscyphus albidus did not cause necrotic lesions on the stems of either Fraxinus species. In contrast, H. fraxineus induced necroses on all inoculated rachises of both ash species with mean lengths of 8.4 cm (F. excelsior) and 1.9 cm (F. pennsylvanica). Necroses also developed on all of the inoculated F. excelsior stems (mean length 18.0 cm), whereas on F. pennsylvanica such lesions only occurred on about 5% of the stems (mean length 1.9 cm). The differences between strains were negligible. No necroses were observed on the control plants. Reisolations of H. albidus were only successful in around 8–11% of the cases, while H. fraxineus was reisolated from 50–70% of the inoculated organs showing necrotic lesions. None of the Hymenoscyphus species were isolated from the control plants. Our data confirm H. fraxineus’ high virulence with regards to F. excelsior and demonstrate a low virulence in relation to F. pennsylvanica under field conditions in Poland. Hymenoscyphus albidus did not express any perceivable pathogenicity on both host species.

Occurrence of Phytophthora plurivora and other Phytophthora species in oak forests of southern Poland and their association with site conditions and the health status of trees

Phytophthora plurivora and other Phytophthora species are known to be serious pathogens of forest trees. Little is known, however, about the presence of P. plurivora in Polish oak forests and their role in oak decline. The aims of this study were to identify P. plurivora in healthy and declining Quercus robur stands in southern Poland and to demonstrate the relationship between different site factors and the occurrence of P. plurivora. In addition, the virulence of P. plurivora and other Phytophthora species was evaluated through inoculations using 2-year-old oak seedlings. Rhizosphere soil was investigated from 39 oak stands representing different healthy tree statuses. The morphology and DNA sequences of the internal transcribed spacer regions (ITS) of the ribosomal DNA and the mitochondrial cox1 gene were used for identifications. P. plurivora, an oak fine root pathogen, was isolated from rhizosphere soil samples in 6 out of 39 stands. Additionally, Phytophthora cambivora, Phytophthora polonica and Phytophthora rosacearum-like were also obtained from several stands. The results showed a significant association between the presence of P. plurivora and the health status of oak trees. Similar relationships were also observed for all identified Phytophthora species. In addition, there was evidence for a connection between the presence of all identified Phytophthora species and some site conditions. Phytophthora spp. occurred more frequently in declining stands and in silt loam and sandy loam soils with pH ≥ 3.66. P. plurivora and P. cambivora were the only species capable of killing whole plants, producing extensive necrosis on seedling stems.

Two new Leptographium spp. reveal an emerging complex of hardwood-infecting species in the Ophiostomatales

Species of Leptographium are generally characterized by mononematous conidiophores and are commonly associated with bark beetles and weevils. These species are responsible for sapstain and in some cases serious diseases on a range of primarily coniferous trees. In comparison with coniferous trees, the occurrence of Leptographium species on hardwood trees has been poorly studied in Europe. During a survey of ophiostomatoid fungi on various tree species in Norway and Poland, three unusual species, which fit the broader morphological description of Leptographium spp., were found in association with Scolytus ratzeburgi, Dryocoetes alni and Trypodendron domesticum on a variety of hardwoods, and from wounds on Tilia cordata. Phylogenetic analyses of sequence data for three gene regions (ITS2-LSU, β-tubulin, and TEF1-α) showed that these Leptographium species are phylogenetically closely related to each other and form a well-supported lineage that included Grosmannia grandifoliae and Leptographium pruni. The first species could be distinguished from the other Leptographium species based on conidiophores arising from spiral hyphae, chlamydospore-like structures and a hyalorhinocladiella-like synanamorph in culture. The second species differs from the previous one by having distinctly shorter conidiophores and smaller conidia. This species also produces a well-developed sporothrix-like synanamorph with denticulate conidiogenous cells. Based on these unusual morphological characteristics and distinct DNA sequences, these fungi were recognised as new taxa for which the names Leptographium trypodendri sp. nov. and L. betulae sp. nov. are provided. The third group of isolates belonged to Grosmannia grandifoliae, representing the first report of this species outside of the USA. The newly defined G. grandifoliae complex is the first species complex in Leptographium s.l. consisting of only hardwood-infecting species.

Identification of sapstain fungi from Scots pine pallets and assessment of their staining ability

Wood-staining fungi, moulds and decay fungi colonize freshly cut wood. Of these, only ophiostomatoid fungi are considered major agents of sapstain on logs and freshly sawn lumber because they can cause permanent staining and discoloration. Sapstain in pine pallets significantly reduces their market value and use in the food industry. The aim of this study was to identify the sapstaining fungi that colonize Scots pine wood used in pallet production. In addition, we evaluated the growth and stain intensity of six isolates of ophiostomatoid fungi on freshly cut Scots pine billets. Fungi were isolated from samples of Scots pine and identified based on morphology and DNA sequence comparisons for three gene regions (ITS, β-tubulin, TEF-1α). A total of 1259 isolates representing 31 fungal species were obtained from the pine samples in Poland during July and September. The isolates represented different ecological and taxonomical groups and belonged to the categories of staining fungi, decay fungi and surface moulds. The most frequently isolated fungi were ophiostomatoid fungi (14 species including an unknown Leptographium sp. and Ophiostoma sp.) and moulds (mainly Trichoderma and Mucor spp.). Of the ophiostomatoid species, Ophiostoma floccosum, O. piceae, O. piliferum and Endoconidiophora pinicola were the predominant species. The results of pine billet inoculation showed that among the six fungal species tested, E. pinicola exhibited the fastest growth in all three directions on the billets. Ophiostoma minus and O. piliferum displayed moderate growth rates in pine billets, while O. floccosum, O. piceae, and Leptographium lundbergii grew very slowly, especially in the tangential and radial directions. The information provided in this paper will help develop more effective control strategies for sapstain prevention in Scots pine.

Diversity of Ophiostomatales species associated with conifer-infesting beetles in the Western Carpathians

Bark beetles (Coleoptera: Curculionidae, Scolytinae) are commonly recognised as important agents of tree mortality in coniferous forests of the Western Carpathians. They, together with weevils, are consistently associated with ophiostomatoid fungi. Information regarding conifer beetle-associated fungi in the Western Carpathians remains incomplete and unreliable, particularly with respect to fir-infesting bark beetles. This study aims to clarify associations between fungi in the genera Graphilbum, Leptographium, Ophiostoma and Sporothrix (Ophiostomatales) and their beetle vectors in Norway spruce (Picea abies), European larch (Larix decidua) and silver fir (Abies alba). Samples associated with 20 bark beetle species and weevils were collected from nine stands in Poland and the Czech Republic. Fungi were isolated from adult beetles and galleries. Isolates were identified based on morphology, DNA sequence comparisons for four gene regions (ITS, LSU, ß-tubulin, TEF 1-α) and phylogenetic analyses. In total, 46 distinct taxa were identified, including 25 known and 21 currently unknown species. Several associations between fungi and subcortical insects were recorded for the first time. In addition, O. borealis and O. quercus were detected from A. alba for the first time. The composition of the fungal communities varied among the studied tree species and to a lesser degree among the beetle species. The spruce-infesting bark beetles were commonly associated with species of Leptographium s. l. and Ophiostoma s. str.; the larch-infesting bark beetles were often associated with Ophiostoma s. str. and Sporothrix, while the fir-infesting bark beetles were commonly associated with Ophiostoma s. str. and Graphilbum. The most commonly encountered fungal associates of the examined insects were (a) Grosmannia cucullata, G. piceiperda, Grosmannia sp. 1, Ophiostoma macroclavatum and O. piceae with the spruce-infesting bark beetles; (b) O. pseudocatenulatum and Sporothrix sp. 1 with the larch-infesting bark beetles; and (c) O. piceae, Ophiostoma sp. 2 and Graphilbum sp. 2 with the fir-infesting bark beetles. The differences in fungal associates among the bark beetle species occurring on P. abies, L. decidua and A. alba could be linked to the different habitats that these beetles occupy.

Distribution of Sceliphron destillatorium Illiger, 1807 (Hymenoptera, Sphecidae) in Poland

There have been 49 confirmed localities of Sceliphron destillatorium Illiger, 1807 in Poland, including 39documented localities and 10 undocumented localities requiring confirmation. Based on the locations of new localities of S. destillatorium as well as those already known from the literature, it has been possible to map where in Poland thisspecies appears as well as the northern limits of its range.

A preliminary study on the pathogenicity of three blue-stain fungi associated with Tetropium spp. to Norway spruce in Poland

Wstępne badania nad patogenicznością trzech grzybów siniznowych związanych z Tetropium spp. na świerku pospolitym w Polsce A preliminary study on the pathogenicity of three blue-stain fungi associated with Tetropium spp. to Norway spruce in Poland To evaluate pathogenicity of some fungi associated with Tetropium spp. (Coleoptera: Cerambycidae): Grosmannia piceiperda, Grosmannia penicillata and Ophiostoma tetropii the inoculation of Norway spruce (Picea abies) trees experiment was performed. The inoculation densities were 400 and 800 points/m2 in a 60 cm wide band at breast height. The effects of inoculation were examined 23 weeks after treatment. Two trees inoculated with G. piceiperda were dying at harvest. Inoculation with G. piceiperda produced significantly greater phloem and sapwood lesions in comparison with other fungi and control. Moreover, the percentage of occluded sapwood was also highest following G. piceiperda inoculations at 800 points/m2. Ophiostoma tetropii, however, produced more occlusions than G. piceiperda at 400 points/m2. These preliminary results suggest that G. piceiperda is pathogenic and may contribute to the mortality of spruces infested by Tetropium beetles.

Three new Leptographium spp. (Ophiostomatales) infecting hardwood trees in Norway and Poland

Species of Leptographium are characterized by mononematous or synnematous conidiophores and are commonly associated with different arthropods. Some of them also produce a sexual state characterised by globose ascomata with elongated necks. Compared to investigations on coniferous trees, the occurrence of Leptographium species on hardwood trees has been poorly studied in Europe. During a survey of ophiostomatoid fungi on various hardwood tree species in Norway and Poland, three unusual species, which fit in the broader morphological description of Leptographium spp., were found in association with Trypodendron domesticum, Trypodendron signatum and Dryocoetes alni, and from wounds on a variety of hardwoods. Phylogenetic analyses of sequence data for six different loci (ITS1–5.8 S–ITS2, ITS2-LSU, ACT, β-tubulin, CAL, and TEF-1α) showed that these Leptographium species are phylogenetically closely related to the species of the Grosmannia olivacea complex. The first species forms a well-supported lineage that includes Ophiostoma brevicolle, while the two other new taxa resided in a separate lineage; possibly affiliated with Grosmannia francke-grosmanniae. All the new species produce perithecia with necks terminating in ostiolar hyphae and orange-section shaped ascospores with cucullate, gelatinous sheaths. These species also produce dark olivaceous mononematous asexual states in culture. In addition, two of the newly described species have a second type of conidiophore with a short and non-pigmented stipe. The new Leptographium species can be easily distinguished from each other by their appearance and growth in culture. Based on novel morphological characters and distinct DNA sequences, these fungi were recognised as new taxa for which the names Leptographium tardum sp. nov., Leptographium vulnerum sp. nov., and Leptographium flavum sp. nov. are provided.

Forest management and hunting in areas adjacent to national parks: the example of the Magura National Park

Abstract the aim of the study was to evaluate arguments for hunting and its impact on forest management and conservation in national parks as well as areas directly adjacent to them. the issue was examined using the example of the Magura national Park (MnP) for which data on the number of deer and predatory mammals included in the statistical yearbooks for 2013-2014 were available. the quality and size of the food sources provided by this type of forest habitat were evaluated using data obtained from the literature. We also included data on the dietary habits of wolves and lynxes as well as their impact on the number of large ungulates in our analysis. the maximum carrying capacity of forest stands in the Magura national Park was determined to be 789 deer units (dear unit = 1 red deer or 0.3 elk or 5 roe deer), whereas in fact in 2014, the abundance of ungulates reached a total of 1230 deer units. our analysis evaluating the impact of wolf and lynx populations on ungulates in the area showed that these predators can kill up to 212 deer per year (140 individuals by wolves and 72 by lynxes). the growth in deer population, however, varies from 25.8% to 27.7%, which in the MnP amounts to 258-277 new born individuals per year, meaning that the wolf and lynx populations in the MnP are not able to prevent the number of deer from growing. the current population of ungulates (1230 deer units ) having reached a density of 6.6 deer units/km2 exceeds the capacity of the MnP and thus poses a real threat to maintaining both, the nature of the park and the adjacent stands. this article shows that the natural maintenance of balance in the predator-prey relationship is unlikely under these conditions and failure to allow for anthropogenic interference to regulate the number of ungulates in protected areas may result in an increase in the density of their population. Potential destruction of other valuable assets such as forest habitats may consequently follow. the current population of ungulates (1,230 deer units) having reached a density of 6.6 deer units/km2 exceeds the capacity of the MnP and thus poses a real threat to maintaining the nature of both the park and the adjacent stands. this article shows that the natural maintenance of balance in the predator-prey relationship is unlikely under these conditions and failure to allow for anthropogenic interference to regulate the number of ungulates in protected areas may result in an increase in the density of their population. Potential destruction of other valuable assets such as forest habitats may consequently follow.