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Somaclonal variation of calli and regenerated plants of spring wheat ware detected by using technique RAPD in the study. Calli at different culture stages and regenerated plants derived from young spikes and immature embryos were used as materials. Molecular variation could be reflected from electrophoresis patternof RAPD fragments at different culture stage in calli, and in regenerated plants derived from different explants, even no phenotype variations were found. Somaclonal variation in calli and in regenerated plants appeared regularly: A higher frequency of variation in hybrids F2 was detected than that of the cultivar that is stable genetically. High variation frequency of RAPD fragments appeared in calli when cultured 75 days. The identical variations of RAPD fragments were observed in calli and in the regenerated plants induced from different genotype or explants. The variation frequency detected is higher in regenerated plants than that of in calli. RAPD could be applied easily and simply to determine variation in level of DNA at each stage cultured in vitro.