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Heavy metal pollution is a major environmental problem all over the world. It is known that high concentration of heavy metals in soils and waters cause genotoxicity and damage to most of the functional biomolecules. This event is a potentially dangerous for every living organism from plants to humans. The aim of this study was to determine the methylcytosine levels in sunflower (Helianthus annuus L.) genome and changes in methylation patterns under heavy metal of zinc stress. Sunflower seeds were treated with different concentrations of zinc (0, 20, 40, 80, 160, 320, 640, 1280ppm) for 3 weeks. The overview of the seedlings showed that the growth and development of seedlings were inhibited by 80ppm and following the increase of concentrations. The genomic DNA was extracted 3 weeks after zinc treatments. DNA band variations were revealed by random amplified polymorphic DNA (RAPD) analysis. And then, levels of methylcytosine in the genome were detected with the technique of coupled restriction enzyme digestion-random amplification (CRED-RA). CRED-RA is a significant technique which uses methylation-sensitive enzymes, HpaII and MspI for determining the methylation patterns. According to the RAPD and CRED-RA analysis, GTS rates adapted with RAPD profiles, when compared, the highest rate was 92.85% at 20ppm zinc concentration. The lowest rate was 83.33% at 640ppm zinc. Additionally, total methylation and full-methylation rates decreased until 160ppm zinc stress and then started to increase at 320ppm and more concentrations of zinc stressin comparison to the control groups. Similarly, the maximum percentage of non-methylation was detected 5.7% at 160ppm zinc stress. According to results of RAPD and CRED-RA analysis, very important polymorphism and methylation changes were observed in sunflower plants subjected to zinc stress.

Determination of DNA Methylation Levels with CRED-RA Technique in the Genome of Sunflower Seedlings (Helianthus annuus L.) Subjected to Zinc Stress