Rhododendron dieback was continuously observed with increasing frequency on Rhododendron maximum (Rosebay Rhododendron) during the last 20 years in the southern Appalachian Mountains. The dieback was especially evident following several years of drought from 2004 to 2008 recorded in Great Smoky Mountains National Park (GRSM). With the concern that a disease epidemic could occur, a holistic study evaluated site factors including tree health, number of clonal units, aspect, slope, depth to bedrock, and rhizosphere microbes. This study was conducted at two locations: Laurel Falls in GRSM and Albert Mountain in Nantahala National Forest (NNF). Yearly sampling for nematodes showed no differences in frequencies across or between years. A total of 11 species were identified from replicated healthy and dieback plots with no significant trends observed. Criconemella xenophus, Helicotylenchus sp., and Meloidogyne sp. were the species most commonly found. Belonolaimus sp. occurred at the NNF site at below 1% of the total nematode population identified, but this nematode species is considered damaging to crops and forest nursery seedlings even at low numbers. Fungal/Oomycota diversity and densities were determined from roots and rhzosphere soil samples using three identification methods. The results ranged from 110 species of fungi to 0 for Oomycota. Of 110 fungi isolated, one putative root pathogen was identified, and the saprophytic species Mycena silvae-nigrae (unknown Basidiomycota 1) was the most common match using the GenBank database. Elevation at NNF was significantly greater than at GRSM, with significantly greater dieback levels at the higher elevation. Furthermore, greater dieback ratings were associated with significantly greater tree diameters. No trends were observed for percent slope or nutrient levels when compared between healthy and dieback sites or locations. Site factors such as aspect, elevation, associated nematode species, and a putative root pathogen may form a disease complex resulting in Rosebay Rhododendron dieback.