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The vegetation in the Qinghai-Tibetan Plateau is thought to be highly sensitive and more vulnerable to global climate change than that of other areas. The uplift of the plateau as well as the climatic oscillations during glacial periods had a profound impact on plant species distribution and genetic diversity there. In the present study, seven pairs of cpSSR (chloroplast Simple Sequence Repeat) primers were utilized to detect genetic varieties of Populus cathayana Rehd populations from their natural range in the southeastern areas of Qinghai-Tibetan Plateau. A total of 28 alleles and 12 different haplotypes were detected. The proportion of haplotype variation among populations (GST = 0.794, NST = 0.900) indicated high level of genetic differentiation among populations and a significant phylogeographic structure (NST > GST, P < 0.05). This appears to support the hypothesis that these populations were derived from multiple refugia areas during the Quaternary climatic oscillations. Based on the haplotype network and mismatch distribution analyses, we found no evidence of postglacial range recolonization and expansion by P. cathayana in this region. This might be mainly due to the complex topography of the southeastern part of the Qinghai-Tibetan Plateau. The lofty mountain ranges and deep valleys in this region might have prevented long-distance migrations of this species during the climatic amelioration.