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Fungi colonising root tips of Pinus sylvestris and Picea abies grown under four different seedling cultivation systems were assessed by morphotyping, direct sequencing and isolation methods. Roots were morphotyped using two approaches: (1) 10% of the whole root system from 30 seedlings of each species and (2) 20 randomly selected tips per plant from 300 seedlings of each species. The first approach yielded 15 morphotypes, the second yielded 27, including 18 new morphotypes. The overall community consisted of 33 morphotypes. The level of mycorrhizal colonisation of roots determined by each approach was about 50%. The cultivation system had a marked effect on the level of mycorrhizal colonisation. In pine, the highest level of colonisation (48%) was observed in bare-root systems, while in spruce, colonisation was highest in polyethylene rolls (71%). Direct internal transcribed spacer ribosomal DNA sequencing and isolation detected a total of 93 fungal taxa, including 27 mycorrhizal. A total of 71 (76.3%) fungi were identified at least to a genus level. The overlap between the two methods was low. Only 13 (13.9%) of taxa were both sequenced and isolated, 47 (50.5%) were detected exclusively by sequencing and 33 (35.5%) exclusively by isolation. All isolated mycorrhizal fungi were also detected by direct sequencing. Characteristic mycorrhizas were Phialophora finlandia, Amphinema byssoides, Rhizopogon rubescens, Suillus luteus and Thelephora terrestris. There was a moderate similarity in mycorrhizal communities between pine and spruce and among different cultivation systems.

Coarse woody debris is important for mycodiversity in forest ecosystems, but its availability in managed stands is reduced. Leaving dead wood during felling is suggested as an option to sustain and restore the diversity. However, little is known what fungi would colonise freshly cut wood left on managed sites, and how the colonisation process is influenced by ecological factors. During summer and autumn, 120 freshly cut Picea abies stem sections over 8 cm in diameter were placed upright in mapped locations over two discrete plots separated by 100 m in a north-temperate forest. After seven weeks the sections were collected, and isolation and identification of fungi was done from their upper surfaces. In all 943 fungal strains were isolated, representing 97 species. Species richness in the summer survey was 42.5% higher than during the autumn survey. Low species similarity characterized the different seasons (Sorensen indices: S(S) = 0.36 and S(N) = 0.34) and for 21 species (22%) observation frequency was significantly affected by season. As a result, community structures in summer and autumn differed notably (z-test; P < 0.001). Species richness between the two plots differed by less than 10%, but there were 64 species (66%) found only in one of them, thus qualitative similarity was low (S(S) = 0.49). Quantitative similarity was higher (S(N) = 0.63), indicating that the dominant species colonised wood to a similar extent in both areas. Fungal community structure differed significantly among the two plots (z-test; P < 0.001). Our data showed that freshly cut CWD contributed to mycodiversity in managed north-temperate forest, providing habitats for numerous individuals from over 100 species. The fungal community within a single stand differed markedly both across small distances and over the seasons. In order to sustain and enhance mycodiversity in managed stands. coarse wood should always be left during harvesting. This study also demonstrates the importance of molecular identification and ITS sequence databases for exploring fungal diversity in natural communities.

The aim of the present work was to determine the identity and molecular relationships between 127 strains of dark septate (DS) fungi isolated from healthy root tips, decayed coarse roots, live healthy-looking stems, coarse (stumps, snags and logs) and fine (tree branches and tops) woody debris in temperate-boreal forests in Sweden and Lithuania. Sequence analysis of ITS rDNA was used to identify the fungi. In a neighbour-joining similarity tree, all sequences were grouped into five distinct clusters. Within each of these, ITS rDNA sequence variation consisted of 2-18 nucleotides, corresponding to 1-3% of their total length. The four least variable clusters were supported with high bootstrap values of 86-100%. Comparisons with the sequences in the GenBank database showed that all our strains had a 95-100% homology with identified Phialocephala species, and they were thus assigned to this genus. The representatives of two clusters were identified, as P. fortinii and P. dimorphospora. The representatives of three remaining clusters were defined as Phialocephala sp. 35, Phialocephala sp. 6 and Phialocephala sp. 18. Within each of these clusters, ITS rDNA sequence uniformity was higher than that observed within P. fortinii and P. dimorphospora. Consequently, their clusters were most discrete, supported with bootstrap values of 100%. Genetic variation in the five distinguished Phialocephala species and their possible ecological roles are discussed. Phialocephala sp. 6 was confined to healthy root tips of conifers. P. dimorphospora was only associated with dead woody tissue of P. abies. P. fortinii, Phialocephala sp. 18 and sp. 35 were isolated from both dead and living conifers and Betula pendula. In conclusion, the present study revealed the ability of fungi from the genus Phialocephala to colonise and persist in live and dead trees under strikingly different ecological conditions.

Seven polymorphic microsatellite markers were developed for the wood-decay basidiomycete Phlebia centrifuga. The primers were identified using two techniques, based on intersimple sequence repeats (ISSR) and amplified fragment length polymorphism (AFLP), respectively. The markers were screened on 27 isolates from Europe and North America. Two markers varied only on a worldwide scale, but not within Europe. The other five showed variation on both scales. These markers will now be used to characterize populations of P. centrifuga, which is red-listed as near-threatened in its natural habitat due to human disturbance. [ABSTRACT FROM AUTHOR]

Spread of Heterobasidion annosum (Fr.) Bref. was investigated in a 60-year-old Norway spruce (Picea abies (L.) Karst.) stand in central Lithuania. Of the 375 trees sampled, H. annosum was isolated from 85 (22.7%). Forty nine (56.3%) of the isolates belonged to the S group and 38 (43.7%) to the P group. Among the S-group isolates, 35 genets occupied 49 trees (1.4 trees per genet on average). In the P group, 24 genets were detected in 37 trees (1.5 trees per genet on average). The largest S and P genets were 21 and 20 m in diameter, respectively. The S group spread was more extensive within stems than for the P group; mean length of the decay column for S-group isolates was 459 1 159 cm (mean 1 SD), and it was 327 1 101 cm for P-group isolates (p = 0.0001). Average stump area affected by decay was 56 1 15%for the S group, and 47 1 16% for the P group (p = 0.016). This is the first published field data on intersterility group related difference in decay extension in tree stems. The S group is better adapted than the P group to growth in Norway spruce wood. [ABSTRACT FROM AUTHOR]