title: ITS sequence R. pseudoacacia
design: The experimental soil was collected from forest development, testing, and demonstration of mixed species reforestation in southern Appalachia to understand soil microbial community characteristics and ecological dynamics. It is a 369-acre farm protected by the Blue Ridge Conservancy containing a 26-acre former Christmas tree plantation with reforested portions at the universitys Blackburn Vannoy Estate campus (37 27 38.3 N, 126 57 07.5 E). As the exclusive owner of these land parcels, Appalachian State University aims to sustainably preserve, restore, and manage these properties. Therefore, understanding soil microbial ecology dynamics is essential for forest restoration on degraded lands (Song et al., 2019), and they are not well-documented in former Christmas tree plantations. Ten soil sampling points were established on the Vannoy property including abandoned pastures, a former Christmas tree plantation, and a forest fragment. At each site, composite samples were collected for DNA extraction, and Illumina-based ITS Miseq sequencing using methods previously used by Ogwu et al. (2019). This method is capable of revealing the status of root-associated ectomycorrhizal and arbuscular mycorrhizal fungi communities and functional interactions. Ashe County where the farm is located experiences a cool montane temperate climate with a mean annual temperature of 32 F and an annual mean precipitation of 50 inches. Sampling sites were selected because they are representative of forest fragments and pasture land/Christmas tree plantation in terms of plant species composition and soil pH/texture characteristics, as well as visible anthropogenic impacts.