Four species of the genus Berberis L. have been reported from different regions, including Khorassan provinces. In this article, a revision of this genus in Khorassan provinces is presented. For better validation of the results, morphological, palynological, chromosomal and molecular studies were conducted on specimens collected from Khorassan provinces and preserved in FUMH and TARI herbaria. Morphological study resulted in recognition of three species of Berberis, from which two species are new records for Khorassan. Four unknown new taxa with new morphological characters were also identified, but left for further analysis because of the high percentage of polyploidy and hybridization in this genus. In order to analyze morphological data, 35 OTUs were provided and scored using the distance method and PCoA. The shape and size of pollen grains were nearly equal in the examined taxa, and there were no obvious differences between them. Chromosomal examination indicated tetraploidy (2n = 4x = 56) in all of the studied taxa. Molecular studies were accomplished by RAPD and sequencing of the ITS region to construct a framework of relationships between the taxa. Molecular studies emphasized the difference in the four unknown taxa from others. The total evidence indicates that Berberis L. shows a high percentage of polyploidy and hybridization.

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