Genetic diversity is vital to the sustainable utilization and conservation of plant species. Rhododendron rex subsp. rex Lévl. is an endangered species endemic to the southwest of China. Although the natural populations of this species are facing continuous decline due to the high frequency of anthropogenic disturbance, the genetic information of R. rex subsp. rex is not yet elucidated. In the present study, 10 pairs of microsatellite markers (nSSRs) and three pairs of chloroplast DNA (cpDNAs) were used in the elucidation of the genetic diversity, population structure, and demographic history of 11 R. rex subsp. rex populations. A total of 236 alleles and 12 haplotypes were found. A moderate genetic diversity within populations (H E = 0.540 for nSSRs, Hd = 0.788 for cpDNA markers), high historical and low contemporary gene flows, and moderate genetic differentiation (nSSR: F ST = 0.165***; cpDNA: F ST = 0.841***) were detected among the R. rex subsp. rex populations. Genetic and geographic distances showed significant correlation (p < 0.05) determined by the Mantel test. The species exhibited a conspicuous phylogeographical structure among the populations. Using the Bayesian skyline plot and species distribution models, we found that R. rex subsp. rex underwent a population demography contraction approximately 50,000-100,000 years ago. However, the species did not experience a recent population expansion event. Thus, habitat loss and destruction, which result in a population decline and species inbreeding depression, should be considered in the management and conservation of R. rex subsp. rex. Plants 2020, 9, 338 2 of 16 potential genetic resources for the development of new cultivars that can adapt to diverse environmental conditions [4]. In addition, plants in the genus Rhododendron L. produce numerous chemical constituents and are recognized as an important source of bioactive phytochemicals [5]. Some Rhododendron species are used as traditional medicine in China, India, Europe, and North America against various diseases, such as inflammation, pain, skin ailments, common cold, and gastrointestinal disorders [5]. However, as an important natural resource for human daily life and ecosystem composition, most Rhododendron species are facing risk of extinction due to the high frequency of anthropogenic disturbance [6]. Thus, research on the population genetic information of Rhododendron species is undoubtedly beneficial for germplasm protection and sustainable utilization [6][7][8][9].Inferring genetic information is recognized as the undisputed basis for the sustainable exploitation and conservation of plant diversity [10,11]. Different molecular markers are used in assessing genetic information and identifying distinct plant populations for management and conservation [12][13][14]. Microsatellite markers (SSRs) are used in revealing the genetic characteristics and related influence factors of plant species at individual and population levels due to their desirable advantages [13,15]. Chloroplast DNA (cpDNA), whic...