Nine samples of Rumex acetosella complex in Bulgaria were studied using ISSR markers. The taxa were defined on the basis of their specific morphological features. Total genomic DNA was extracted from fresh leaves and used as templates for ISSR - PCR. Six ISSR primers from the collection of the University of British Columbia (Nucleic Acid-Protein Service Unit, UBS Primer Set #9) were used to amplify polymorphic microsatellite loci. The PCR reactions were carried out in Thermal Cycler 2720 (Applied Biosystems). To obtain uniformity in PCR reactions we used PCR master mix (Fermentas, Cat No K0171). The amplified unambiguous bands were scored by molecular weights and redistributed in classes to compile a presence/absence matrix. The dendrograms were based on the results obtained by each of primers used. Cluster and principle analysis showed that the population can not be separated significantly. There is not relationship between morphological distribution and genetic distance of R. tenuifolius and R. acetosella. We suggest the distribution and relationship between members of this group require further study and reassessment of the taxonomic status.
Testing of ISSR markers in Rumex Acetosella complex in Bulgaria