Methodology

Data Collection

Field sampling sites (n=6) were set up in dry prairie grassland and parkland ecoregions within Alberta, Canada. A modified field sampling method from Kumpula (2020) and Edwards (2018) was designed (Fig.2) using a 1 m x 0.8 m quadrat placed in a nested design to accommodate the heterogenous community within a site. Sample sites were selected to cover two distinct habitats, trying to cover a distinct plant assemblage for the forested and grassland ecoregions. There were three sites established in a semi-arid natural region in a prairie grassland, with mixed grass species, short shrub and biocrust communities (Government of Alberta and Alberta Tourism, Parks and Recreation 2015). The other half of the sites were established in a hemi-boreal parkland ecoregion, with mixed forbs and woody species as the dominant vegetation. Field experts conducted time unlimited surveys at each of the 9 quadrats at a site identifying plants using ID keys. Species lists were collected for each plot with percent cover taken for each quadrat (Fig. 3). Community DNA was collected at the same quadrats. A small 0.5 cm x 0.5 cm section was taken and frozen within 4 hours and then stored at -80 C until extraction. Lab processing occurred at Innotech Alberta, extracting genomic DNA and then amplifying common barcode segments (rbcL and matK) before sequencing using the native metabarcoding protocol (SQK-NBD114.96) using Oxford Nanopore Next generation sequencing. 

Figure 4: Site photos of the six sites where data was collected. a-c) were collected in a forested ecoregion and d-f) were collected in a mixed grass prairie. 

Figure 5: Maps of sites that were selected in two ecosites. a) Parkland Hemi-Boreal habitats near Edmonton Alberta and b) short and mixed grass native prairie near Brooks Alberta.