Effects Of Curcumin And Decitabine On The DNA Methylation And Development Of Chrysanthemum

Epigenetic and genetic change can lead to changes in morphology and growth of animals and plants. In this study, we analyzed the alterations in levels and patterns of DNA methylation in chrysanthemum after being treated separately by two kinds of demethylation inhibitors curcumin and decitabine(5-Aza-2'-deoxycytidine) using two methods:(1) Methylation sensitive amplification polymorphism(MSAP) technique and(2) cDNA amplified fragment length polymorphism(cDNA-AFLP). At the same time, we compared the effects of demethylation inhibitor in two ways: the tissue culture and smearing, in order to find out a more economic and efficient way for practice. The results are as follows:(1) The ‘Jinba' chrysanthemums were treated by curcumin with different concentrations(0 ?mol/L, 100 ?mol/L, 200 ?mol/L, 400 ?mol/L) in tissue culture for 30 days. Methylation sensitive amplification polymorphism(MSAP) analysis showed that its DNA methylation rates were respectively 58.40%, 50.44%, 48.24% and 46.00%. The height reduced 0.82 cm, 1.08 cm and 1.35 cm; the number of multiple shoots reduced 0.41, 0.75, 1.48; and the number of leaves reduced 6.00, 9.00 and 13.44. The yellowing leaves gradually recovered after transplanting, but the inhibitory effects on DNA methylation persisted. The DNA methylation ratio experienced a greater decline when the concentration increased in the tissue culture than by using smearing to treat the chrysanthemums.(2) The results of two different treatment methods: the DNA methylation levels changed more when the chrysanthemums were treated in tissue culture than by smearing. A multitude of treatments were infected by aphid. The concentration of 200 ?mol/L curcumin had the best effect among all concentrations, which led to an earlier flowering of ‘Jinba' chrysanthemums by 5-8 days.(3) cDNA-AFLP analysis of the chrysanthemums treated in tissue culture showed that 26, 38, 4 transcript derived fragments(TDFs) were respectively obtained in the 100 ?mol/L, 200 ?mol/L, 400 ?mol/L treatment. No significant positive correlation with the concentration.(4) The heights were respectively 3.57 cm, 2.95 cm, 2.99 cm and 1.71 cm; the number of multiple shoots were respectively 3.91, 3.25, 2.83 and 1.83; and number of leaves were respectively 30.83?27.08?20.58?16.66 when the chrysanthemums were treated with different concentrations(0 ?mol/L, 50 ?mol/L, 100 ?mol/L, 150 ?mol/L) of decitabine in tissue culture for one month. Methylation sensitive amplification polymorphism(MSAP) analysis of tissue culture treatment showed that its DNA methylation rates were respectively 51.16%, 49.57%, 48.18%, 43.54% while in the smeared groups, DNA methylation rate were 51.60%, 48.43%, 46.54%, 42.78%. The concentration of 100?mol/L decitabine made the flowering time of Zijinglin chrysanthemums ahead of 3-5 days by smearing method, and a multitude of treatments were infected by aphid.