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



E. Bolukbasi, E. S. Aras



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. 



10.22161/ijeab/1.3.20



http://ijeab.com/upload_document/issue_files/20%20IJEAB-SEP-2016-28-Determination%20of%20DNA%20Methylation%20Levels%20with%20CRED-RA%20Technique%20in%20the%20Genome%20of%20Sunflower%20Seedlings%20(Helianthus%20annuus%20L.)%20Subjected%20to%20Zinc%20Stress.pdf



http://ijeab.com/submit-paper/

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