Verbascum tzar-borisii (Scrophulariaceae) is a highly endangered plant species, with only two small and isolated populations in Bulgaria. Its present distribution range is highly fragmented, and the size of its populations was dramatically reduced during the past years. We used molecular Inter Simple Sequence Repeat (ISSR) markers to characterize the genetic diversity of this local endemic. Eleven ISSR primers selected from 25 primers gave rise to 110 amplified loci, of which 106 (96%) were polymorphic. We found a low genetic diversity at the population level and a significant genetic differentiation between the investigated populations. Gene flow (Nm) was estimated to be 0.462, indicating a low migration rate between the populations. Analysis of molecular variance revealed that about two-thirds of the genetic variation was partitioned within populations. The high number of private alleles suggests that the extant populations of V. tzar-borisii need to be conserved.
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