Arboriculture & Urban Forestry 42(1): January 2016 software (Applied Biosystems) and a custom 2 bp binning panel (Rees et al. 2004) with a minimum peak height detection of 50 fluorescent units. Microbial Biomass and Microbial Community Lipid Analysis (PLFA/FAME) Lipid analyses were completed using a hybrid procedure of phospholipid fatty acid (PLFA) and fatty acid methyl ester (FAME) analysis (Balser 2001) at the University of Wisconsin–Madison. Approximately 3.0 g of milled, freeze-dried soil was extracted and analyzed according to Wal- drop and Firestone (2004). The bacterial fatty acid standards 9:0 (nonanoic methyl ester) and 19:0 (nonadecanoic methyl ester) (Sigma, St. Louis, Missouri, U.S.) with MIDI peak identification soſt- ware (MIDI, Newark, Delaware, U.S.) were used to identify peaks. Fatty acids were excluded if pres- ent in less than 3 samples or if less than 0.5 mol%. Litter Decomposition Fifteen in situ litter decomposition bags contain- ing approximately 1.0 g of air-dried white oak (Quercus alba) leaves were buried at randomly selected locations in each study site. The litter bags were constructed of gray 1.0 mm fiberglass screen (25 cm × 20 cm) and inserted vertically in the upper 25 cm of the soil profile. Prior to installation, approximately 5 mg of leaf tissue was removed from each litter bag and analyzed for initial total C, total N, and C:N ratio using an NC 2100 Soil analyzer (Alantech, Atlanta, Geor- gia, U.S.) with Clarity 6.12.2006 software (2006 DataApex Ltd, Czech Republic). Three buried litter decomposition bags from each property were removed 10, 30, 60, 90, and 120 days after installation. The bags were washed in deionized water, to remove soil particles, and air dried. The remaining leaf material was pulverized and ana- lyzed for total C, total N, and C:N ratio. Between sampling period changes in total C, total N, and C:N ratio of transposed litter were analyzed using repeated measures ANOVA in statistical analysis software (SAS v9.2, Cary, North Carolina, U.S.). Differences among land uses within a sample period were analyzed in SAS using ANOVA with Duncan’s multiple range test (Duncan’s MRT). 61 Statistical Analysis One-way analysis of variance (ANOVA, α = 0.05) and Duncan’s MRT were conducted using SAS to compare soil physical and chemical proper- ties among land uses. All physical and chemical attributes were normalized according to Clarke and Gorley (2006). The physical and chemical properties of each sample were compared with a Euclidian distance matrix using Plymouth Routines in Multivariate Ecological Research v6 (PRIMER v6, Lutton, UK) (Clarke and Gorley 2006). Analysis of similarities (ANOSIM) and non-metric multi-dimensional scaling (MDS) (2D stress limit of <0.20) with cluster overlay (PRIMER v6) were used to make inferences regarding differences in the edaphic properties of the soils among land uses. Pairwise land-use comparisons were evaluated on the edaphic properties if the global R value was significant (α = 0.05, 999 permutations, Clarke and Warwick 2001). Pairwise R values below 0.50 indicated edaphic variables within the land-use categories were not separated, values between 0.50 and 0.70 were overlapping, and values above 0.70 were well separated (Clarke and Warwick 2001). Estimates of total microbial biomass [sum of total microbial lipids (Balser 2001)] in each land use were compared using a one-way ANOVA (α = 0.05). Individual lipids were standardized by divid- ing the respective lipid abundances (µmol/g soil) by the total lipid abundance within the sample (Balser 2001). Individual peaks within the TRFLP bacterial and fungal profiles were standardized by dividing by the sum of all peak heights within that profile (Rees et al. 2004). Simpson diversity, Shannon diversity, Pielou’s evenness, and Mar- galef species richness (Clarke and Gorley 2006) were calculated from the standardized TRFLP and PLFA/FAME profiles (PRIMER v6). One- way ANOVA (α = 0.05) and Tukey’s Honestly Significant Difference (Tukey’s HSD) were used to evaluate differences in biodiversity measures among land uses for the samples. For multivariate analysis, standardized TRFLP and PLFA/FAME profiles were square-root transformed and ana- lyzed with a Bray-Curtis similarity matrix using ANOSIM and MDS (2D stress <0.20) with clus- ter overlay in PRIMER v6 (Rees et al. 2004). ©2016 International Society of Arboriculture
January 2016
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