Armillaria species in coniferous stands

Identification of the Armillaria species in selected coniferous stands (Scots pine stands, Norway spruce stands and fir stands) was the aim of the work carried out on the basis of mating tests and consideration of macroscopic traits of fruit-bodies. One species of Armillaria [A. ostoyae (Romagnesi) Herink] was found in Scots pine stands, three species [A. ostoyae, A. cepistipes Velenovský and A. borealis Marxmüller et Korhonen] were found in Norway spruce stands and two species [A. ostoyae and A. cepistipes] were found in fir stands

Fibroporia gossypium in northeastern Poland – a preliminary study

A Fibroporia gossypium (Speg.) Parmasto fruit-body was found on a Norway spruce [Picea abies (L.) Karst.] stump in the northeast of Poland (Waliły forest district). The mycelium from wood samples was sequenced (KF725876), identified and used to examine: (i) linear growth of the mycelium on malt-extract medium and (ii) the decay of spruce wood samples. We observed slow mycelium growth (84 mm colony diameter after 28 days). After the first 3 months of wood decay, the average loss of dry mass was 6.7%. After next 3 months, a further loss of 15.4% (the average loss) was recorded until finally 35.98% loss (the maximum loss of wood for a single sample) occurred. Fibroporia gossypium can be considered as a protective biological agent against root rot in threatened Norway spruce stands

Determination of Pleurotus abieticola

The effect of Phlebiopsis gigantea treatment in control of Heterobasidion parviporum in Norway spruce is less effective than that in control of Heterobasidion annosum in pine. It is necessary to apply other fungi, for example, Pleurotus abieticola in Norway spruce stands. Thus, it is necessary to assess the activity of major ligninolytic enzymes, that is, laccase, lignin peroxidase (LiP), manganese peroxidase (MnP) and versatile peroxidase (VP) produced by P. abieticola, which may be effective in the fast degradation of Norway spruce wood. Three strains of P. abieticola (Pa1-3) were grown on pieces of Norway spruce sapwood and heartwood for 50 days in laboratory conditions. Enzymatic activity was determined using spectrophotometry. Pleurotus abieticola produced laccase, LiP, MnP and VP. The activity of laccase was low, ranging 0–3.696 and 0–0.806 mU/μg of protein in sapwood and heartwood, respectively. The highest activity in Pa1 = 3.696 mU/μg of protein in sapwood and in Pa3 = 0.806 mU/μg of protein in heartwood was observed after 30 and 50 days of incubation, respectively. The activity of LiP was also low, ranging 0–0.188 and 0–0.271 mU/μg of protein in sapwood and heartwood, respectively. The highest activity in Pa1 = 0.271 mU/μg of protein in sapwood and in Pa2 = 0.188 mU/μg of protein in heartwood was observed after 40 and 20 days of incubation, respectively. The activity of MnP ranged 0–17.618 and 0–12.203 mU/μg of protein in sapwood and heartwood, respectively. This enzymatic activity peaked at the 50th day of culture on sapwood for the Pa3 strain (17.618 mU/μg of protein) and at the 20th day of culture on heartwood for the Pa1 strain (12.203 mU/μg of protein). The activity of VP with manganese-oxidising properties was found to be high in all strains of P. abieticola, ranging 0–39.19 and 0–59.153 mU/μg of protein in sapwood and heartwood, respectively, whereas the activity of VP with guaiacol-oxidising properties was very low for all P. abieticola strains, ranging 0–0.248 and 0–0.225 mU/μg of protein in sapwood and heartwood, respectively. The values of released hydroxyphenols in P. abieticola strains ranged 24.915–139.766 and 25.19–84.562 µg of protocatechuic acid/ml in sapwood and heartwood, respectively. The values of released methoxyphenols for the evaluated strains of P. abieticola ranged 7.225–23.789 and 1.953–20.651 µg of vanillic acid/ml in sapwood and heartwood, respectively. Further studies with a higher number of strains of this species as well as an optimisation of conditions for the measurement of ligninolytic activity are needed

Determination of Pleurotus abieticola ligninolytic activity on Norway spruce wood

The effect of Phlebiopsis gigantea treatment in control of Heterobasidion parviporum in Norway spruce is less effective than that in control of Heterobasidion annosum in pine. It is necessary to apply other fungi, for example, Pleurotus abieticola in Norway spruce stands. Thus, it is necessary to assess the activity of major ligninolytic enzymes, that is, laccase, lignin peroxidase (LiP), manganese peroxidase (MnP) and versatile peroxidase (VP) produced by P. abieticola, which may be effective in the fast degradation of Norway spruce wood. Three strains of P. abieticola (Pa1-3) were grown on pieces of Norway spruce sapwood and heartwood for 50 days in laboratory conditions. Enzymatic activity was determined using spectrophotometry. Pleurotus abieticola produced laccase, LiP, MnP and VP. The activity of laccase was low, ranging 0–3.696 and 0–0.806 mU/μg of protein in sapwood and heartwood, respectively. The highest activity in Pa1 = 3.696 mU/μg of protein in sapwood and in Pa3 = 0.806 mU/μg of protein in heartwood was observed after 30 and 50 days of incubation, respectively. The activity of LiP was also low, ranging 0–0.188 and 0–0.271 mU/μg of protein in sapwood and heartwood, respectively. The highest activity in Pa1 = 0.271 mU/μg of protein in sapwood and in Pa2 = 0.188 mU/μg of protein in heartwood was observed after 40 and 20 days of incubation, respectively. The activity of MnP ranged 0–17.618 and 0–12.203 mU/μg of protein in sapwood and heartwood, respectively. This enzymatic activity peaked at the 50th day of culture on sapwood for the Pa3 strain (17.618 mU/μg of protein) and at the 20th day of culture on heartwood for the Pa1 strain (12.203 mU/μg of protein). The activity of VP with manganese-oxidising properties was found to be high in all strains of P. abieticola, ranging 0–39.19 and 0–59.153 mU/μg of protein in sapwood and heartwood, respectively, whereas the activity of VP with guaiacol-oxidising properties was very low for all P. abieticola strains, ranging 0–0.248 and 0–0.225 mU/μg of protein in sapwood and heartwood, respectively. The values of released hydroxyphenols in P. abieticola strains ranged 24.915–139.766 and 25.19–84.562 µg of protocatechuic acid/ml in sapwood and heartwood, respectively. The values of released methoxyphenols for the evaluated strains of P. abieticola ranged 7.225–23.789 and 1.953–20.651 µg of vanillic acid/ml in sapwood and heartwood, respectively. Further studies with a higher number of strains of this species as well as an optimisation of conditions for the measurement of ligninolytic activity are needed

Fungi isolated from shoots showing ash dieback in the Wolica Nature Reserve in Poland and artificially inoculated seedlings with Hymenoscyphus fraxineus

Ash dieback caused by an alien, invasive fungus Hymenoscyphus fraxineus is a serious disease of European ash species in many parts in Europe. In Poland, the disease was recorded in the beginning of the 1990s. This study was performed in 2016–2017 with the aim to identify fungi isolated from ash shoots showing dieback symptoms in the Wolica Nature Reserve in Poland, as well as from shoots of two years-old ash seedlings inoculated with H. fraxineus in the greenhouse. The most frequently isolated fungi from shoots of common ash (associated with the pathogenic fungus H. fraxineus) were identified on the basis of sequencing of the internal transcribed spacer region (ITS1) of fungal rDNA. In total, 19 fungal taxa were identified for ash shoots as follows: H. fraxineus, Fusarium avenaceum, Alternaria spp., Phomopsis oblonga, Diplodia mutila and other Phomopsis spp. The pathogen H. fraxineus was not found for all the shoots samples; one year after inoculation the aforementioned fungi and other species as: Alternaria alternata, Bionectria ochroleuca, Epicoccum nigrum, F. acuminatum, F. avenaceum, and Paraphaesphaeria neglecta were identified in inoculation point, as well as H. fraxineus. The same quantitative and qualitative changes of organisms were observed in the case of artificially colonised ash seedlings in the greenhouse, as well as in the shoots of adult ash trees in the forest

Fungi isolated from shoots showing ash dieback in the Wolica Nature Reserve in Poland and artificially inoculated seedlings with Hymenoscyphus fraxineus

Ash dieback caused by an alien, invasive fungus Hymenoscyphus fraxineus is a serious disease of European ash species in many parts in Europe. In Poland, the disease was recorded in the beginning of the 1990s. This study was performed in 2016–2017 with the aim to identify fungi isolated from ash shoots showing dieback symptoms in the Wolica Nature Reserve in Poland, as well as from shoots of two years-old ash seedlings inoculated with H. fraxineus in the greenhouse. The most frequently isolated fungi from shoots of common ash (associated with the pathogenic fungus H. fraxineus) were identified on the basis of sequencing of the internal transcribed spacer region (ITS1) of fungal rDNA. In total, 19 fungal taxa were identified for ash shoots as follows: H. fraxineus, Fusarium avenaceum, Alternaria spp., Phomopsis oblonga, Diplodia mutila and other Phomopsis spp. The pathogen H. fraxineus was not found for all the shoots samples; one year after inoculation the aforementioned fungi and other species as: Alternaria alternata, Bionectria ochroleuca, Epicoccum nigrum, F. acuminatum, F. avenaceum, and Paraphaesphaeria neglecta were identified in inoculation point, as well as H. fraxineus. The same quantitative and qualitative changes of organisms were observed in the case of artificially colonised ash seedlings in the greenhouse, as well as in the shoots of adult ash trees in the forest