Four species of genus Berberis L. have been repoted from different regions, including khorrassan provinces. In this paper 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 plants 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 recognized, and left for more suitable analysis, due to high percentage of polyploidy and hybridization in this genus. In order to analyze morphological data, 35 OTUs were provided and separated using distance matrix and PCoA. The shape and size of pollen grains were nearly equal in studied taxa and there were not 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 ITS region to make a framework of relationships between the taxa. Molecular studies emphasized on difference of four unknown taxa from others. Total evidences indicate that Berberis L. showing high percentage of polyploidy and hybridization

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