Methylation patterns of two repetitive DNA sequences in tobacco tissue cultures and their regenerants

DNA methylation of two repetitive sequences in tobacco nuclear genome was studied in the course ofin vitro dedifferentiation and differentiation. Using 5-mC sensitivè restriction enzymes and DNA/DNA hybridization with 25S-rDNA probe it has been shown that during the early phase of callus induction prominent changes in the methylation pattern occur which are stably maintained during subsequent callus growth. The following protoplast recovery and plant regeneration have again displayed some more modifications of the methylation status. Comparing the patterns of R₀ plants with the original plant material and the calli it can be assumed that both share in the resulting methylation status. The experiments analyzing the HRS60 family of non-transcribed highly repetitive sequences have displayed a quite monotonous methylation status thus indicating no random methylation perturbations in silent DNA sequences.