Contents lists available at ScienceDirect Ecological Indicators journal homepage: www.elsevier.com/locate/ecolind Research paper Selecting fungal disturbance indicators to compare forest soil profile re- construction regimes Nicole Sukdeo ⁎ , Ewing Teen, P. Michael Rutherford, Hugues B. Massicotte, Keith N. Egger Natural Resources and Environmental Studies Institute, University of Northern British Columbia, 3333 University Way, Prince George, British Columbia, V2N 4Z9, Canada ARTICLE INFO Keywords: Soil reclamation Fungal community ITS2 sequencing Disturbance Indicator ABSTRACT Physical disturbance of soil profiles, even at shallow depths, is a ubiquitous consequence of anthropogenic landscape modification, with short-term impacts on important ecological guilds of fungi. DNA-based methods for surveying community composition are widely incorporated into studies attempting to explain fungal responses to forest eco- system disturbances. Here we compare fungal community composition between three distinct soil profile manip- ulations (20 cm depth) and undisturbed control plots in a sub-boreal spruce forest in the Central Interior region of British Columbia, Canada. Fungal community composition differences were tracked by internal transcribed spacer 2 (ITS2) amplicon sequencing, with comparisons drawn using genus-level annotations. Non-metric multidimensional scaling (NMDS) analysis indicated that disturbed-sample community compositions were similar to controls at 0- months and distinct from controls at 5- and 12-months post disturbance, but did not indicate clustering of samples according to disturbance regime. We used Linear Discriminant Effect Size (LEfSe) analysis to identify fungal genera that consistently indicate disturbed or undisturbed (control) treatments across 5- and 12-month sampling times. Four fungal genera (Exophiala, Hyphodontiella, Mastigobasidium, and Umbelopsis) were detected with higher ranges of re- lative abundance in all disturbance regimes when compared to control plots. Deliberate mixing of LFH into mineral horizon soils stimulated multiple genera that were more frequently detectable in replicate plots at 12 months, when compared to undisturbed and immediately re-assembled plots. Four ectomycorrhizal genera (Amphinema, Cortinarius, Piloderma, and Russula) were identified as strong indicators of control plot soils. A single genus, Capronia, was identified as differentially abundant between stockpiled LFH and immediately replaced LFH. Our results are con- sistent with declines in ectomycorrhizal fungal abundance and increases in saprotroph abundance previously re- ported in DNA-based community profiling studies of forest soil disturbance. This investigation demonstrates that bulk soil sampling can be used to evaluate soil-handling regimens to understand fungal community disruption/recovery and highlights LEfSe as an approach to indicator selection in DNA-dependent biodiversity surveys. 1. Introduction Biodiversity monitoring relies on accurate observation of organisms at varying taxonomic levels of classification over large geographical areas. Traditional approaches to monitoring fungal populations in forest ecosystems have focused on macrofungal surveys of fruiting bodies or belowground ectomycorrhizal (EcM) root tips (Geml et al., 2014; Halme et al., 2012). This approach misses fungi that develop fruiting bodies infrequently or not at all, leading to inaccurate or patchy occurrence/relative abundance data (Halme et al., 2012). Metabarcoding-based surveys exploit sequences of target DNA regions (coding or intergenic) that have sufficient levels of intraspecific diversity to discriminate fungal organisms at phylum down to species level (Cristescu, 2014; Thomsen and Willerslev, 2015). The internal transcribed spacer 2 (ITS2) region has been widely used for profiling fungal commu- nity composition (Lindahl et al., 2013) in forest soils studies that have focused on vertical distributions in soil profiles (Baldrian et al., 2012; Lindahl et al., 2007; Santalahti et al., 2016), response to ecosystem dis- turbances (Glassman et al., 2015; Pec et al., 2016; Stursová et al., 2014; Sun et al., 2015), and linking organic matter cycling to fungal taxonomic classifications (Bödeker et al., 2016; Purahong et al., 2016; Talbot et al., 2014; Treseder et al., 2016). DNA samples can be isolated from a variety of sample materials (fruiting bodies, soils, water, ectomycorrhizal root tips, etc.), and the availability of public, curated sequence databases enable broad taxonomic detection by metabarcoding approaches. Further, these attributes also make DNA-metabarcoding an appealing option in biodi- versity monitoring contexts for interpreting fungal community inventories according to functional guild associations. For example, ITS2 sequencing http://dx.doi.org/10.1016/j.ecolind.2017.09.021 Received 30 May 2017; Received in revised form 8 September 2017; Accepted 11 September 2017 ⁎ Corresponding author. E-mail addresses: Nicole.Sukdeo@unbc.ca (N. Sukdeo), ewing.teen@gmail.com (E. Teen), Mike.Rutherford@unbc.ca (P.M. Rutherford), Hugues.Massicotte@unbc.ca (H.B. Massicotte), Keith.Egger@unbc.ca (K.N. Egger). Ecological Indicators 84 (2018) 662–682 Available online 10 October 2017 1470-160X/ © 2017 Elsevier Ltd. All rights reserved. MARK