Evaluation of somaclonal variation during repetitious subcultures of tobacco (Nicotiana tabacum L.) callus

In this study for the purpose of evaluating the somaclonal variation during repetitious subcultures of tobacco (Nicotiana tabacum cv. chopogh), we cultured the leaf and stem explants of tobacco plants in a medium containing 4 mg/l 2,4‐dichlorophenoxyacetic acid and 0.5 mg/l kinetin. After the growth of calluses, they were sub‐cultured in the same medium. After the fourth subculture, calluses were transferred to regeneration MS medium containing hormone concentrations 4mg/l kinetin, 1 mg/l zeatine. After regeneration of seedlings, total genomic DNA was extracted from the primary and regenerated plant. Somaclonal variation of the samples was analyzed using 20 Random RAPD Primer. The electrophoresis pattern of 3 Random Primers including OPC‐09, OPR‐12, OPA‐10 indicated the polymorphism in amplified DNA band. This polymorphism resulted from production of somaclonal variation during subcultures of tobacco callus.