Ulmus glabra Hudson, or Wych elm, occurs as fragmented and relict natural populations in the Central System, which acts as a refugium in the Iberian Peninsula. Considering the importance of the Central System populations of U. glabra, the main objective was to assess their genetic diversity using nuclear microsatellite markers. A total of 360 different genotypes were detected in the 427 U. glabra individuals analyzed. Wych elm populations showed a highly significant genetic differentiation (24%; p = 0.0001). Of the 22 populations studied, population of Rozas de Puerto Real (ROZ) showed the highest values of effective number of alleles (2.803), mean Shannon's diversity (1.047) and expected heterozygosity (0.590). Populations of ROZ and Mombeltrán (MOM) showed the highest values of observed heterozygosity (0.838 and 0.709, respectively), and highly negative values for inbreeding coefficient (−0.412 and −0.575, respectively). Also, most of putative hybrids (50 of 55) were observed in these two populations. Demographic analysis revealed signals for recent (four populations) and ancestral (fifteen populations) bottlenecks. Fragmented populations with diminishing number of individuals, along with anthropogenic intervention and Dutch elm disease (DED), are the main threats to U. glabra populations. From a future perspective, the information generated can be considered in the formulation of conservation strategies for U. glabra populations in the Central System.

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