STUDY OF THE EXPLANTS FROM TISSUE CULTURE FOR EVALUATION OF SOMACLONAL VARIATION USING RAPD MARKERS

Isfahan Azayesh cultivar known as apple dwarfing-rootstock was micropropagated on MS media and under tissue culture condition. 'Azayesh' sucker 4 genomic DNA as control with 5 new seeding derived from tissue culture conducted to subculture 5 and extracted by modified Murry and Thompson's miniprep (1990). In order to examine somaclonal variation through subculturing, RAPD marker was applied. One hundred random primers, produced on extracted DNA in PCR reaction of which ten primers produced polymorphic bands. Selected primers totally amplified ninety-seven bands in all samples which among them, sixty-three showed polymorphism. Cluster analyze of cultivar and genotype performed based on Daies similarity and UPGMA approach. Cluster analyze showed that seedling derived from tissue culture has difference at DNA laved and obtained explants clustered in distinct groups. To come into view that RAPD marker is a suitable and effective tool in order to distinguish and make a distinction of explants which during tissue culture during several subculturing steps and affected by somaclonal variation.