h Maissau Arable field ITS/LSU 96 19 20.4 ± 3.1 92.8 2.33 7.37 Niederschleinz Arable field ITS/LSU 92 34 51.3 ± 12.0 Alvocidib order 66.3 3.27 28.09 Purkersdorf Arable field ITS/LSU 94 32 44.9 ± 9.5 71.3 3.18 23.76 Riederberg Grassland ITS/LSU 92 31 41.4 ± 7.1 77.3 2.84 10.76 Tulln Arable field ITS/LSU 89 24 32.9 ± 8.0 72.9 2.84 15.48 Sourhope (UK)a Grassland SSU 53 18 47.8 ± 22.4 37.7 1.93 3.62 Sourhope (UK)a Grassland ITS 45 22 51.3 ± 20.5 42.9 2.53 7.50 Cristalina (BRA)a Arable field (Soy) SSU 104 22 30.9 ± 7.6 71.2 1.87 2.87 aData for the soils “Sourhope” from the Sourhope Research Station in Scotland, UK (Anderson et al. 2003) and “Cristalina” from the district Cristalina in Goiás, Brazil (de Castro et al. 2008) were taken from the respective publications
bLibrary indicates on which region from rRNA-encoding cluster profiling of the fungal community was done cClones: number of INCB018424 supplier analysed clones for each soil; dSobs: number of observed species in the clone libraries; eChao2 ± SD: Estimated species richness ± standard deviation for the sampling site https://www.selleckchem.com/products/S31-201.html based on the Chao2 richness estimator (Chao 1987) implemented in EstimateS 8.2; f% Cov.: Estimated coverage of the libraries based on observed and estimated species richness; gShann.: Shannon Diversity Index hSimp.: Simpson Diversity Index UniFrac was used to compare the phylogenetic structures of the fungal communities from soils M, N, P, R and T (Lozupone
et al. 2006). To this end sequences were aligned with the ClustalW algorithm in MEGA4 (Tamura et al. 2007), and a neighbor-joining tree was
calculated from the aligned partial LSU sequences. The ITS-region was excluded, since it cannot be unambiguously aligned over such a broad phylogenetic distance. Sequences from an unknown eukaryote (NG_R_F10, Acc. Nr. GU055695) and from a fungus of uncertain affiliation (NG_R_F02, Acc Nr. GU055690) from site R were used as outgroups and excluded from further analyses. Data were weighted for abundance and normalized for branch length for calculating the UniFrac metric of the distance between each pair of soil samples (Lozupone et al. 2006). Results Soil characteristics of the five soils used in the present study are given in Inselsbacher et al. (2009). All soil parameters are within the range for typical arable land as used for cultivation of barley in this area. Fungal communities were Celastrol analysed by direct amplification of fungal ITS/partial LSU regions with primer pair ITS1F and TW13. Cloned PCR products from each soil were grouped by RFLP and up to four representatives from each RFLP type were sequenced. By this approach even closely related sequences (e.g. four different Tetracladium species from soil P with a maximum sequence difference of 3.7%) could be dissected. While the ITS region provides excellent resolution down to the species level, the partial LSU region provides good resolution at higher taxonomic levels when sufficiently identified ITS reference data in public databases are missing (Urban et al. 2008).