机械伤害处理橡胶树萌条树皮的DNA甲基化分析

DNA Methylation Analysis of Barks of Epicormic Shoots Treated With Mechanical Wounding in Rubber Trees

该研究采用甲基化敏感扩增多态性技术,分析了机械伤害处理橡胶树萌条树皮的DNA甲基化的变化。结果显示:(1)与对照相比,伤害后0.5和2h,DNA甲基化水平略有上升;伤害后48h的DNA甲基化水平出现了较大幅度的下降。(2)甲基化变化类型分析表明,在伤害2h主要发生了DNA的甲基化;伤害后48h,主要发生了DNA的去甲基化。(3)差异甲基化位点的回收、测序及注释表明,ATP合酶F1亚基1、磷酸核糖胺-甘氨酸类连接酶、冷激结构域蛋白3、光系统II 47kD蛋白、E3泛素蛋白连接酶RING1、NADH泛醌氧化还原酶和一些假定蛋白参与了伤害的响应。(4)经重亚硫酸盐测序验证,ATP合酶F1亚基1和NADH泛醌氧化还原酶的CCGG位点发生了去甲基化。研究推断DNA的甲基化可能参与了橡胶树萌条对机械伤害的响应。

In the present study, the changes of DNA methylation in the barks of epicormic shoots which treated with mechanical wounding in rubber tree were analyzed by methylation-sensitive amplification polymorphism (MSAP). The results showed that:(1) in comparison with the control, the levels of DNA methylation were slightly increased at the time point of 0.5 and 2 hours after mechanical wounding. And the larger decrease of DNA methylation levels were observed at 48 hours after mechanical wounding.(2)The main DNA methylation and demethylation events were occurred at 2 and 48 hours, respectively.(3)ATP synthase F1 subunit 1, phosphoribosylamine-glycine ligase-like, cold shock domain-containing protein 3, photosystem II 47 kD protein, E3 ubiquitin-protein ligase RING1, NADH-ubiquinone oxidoreductase and some hypothetical protein were identified from the differentially methylated sites induced by mechanical wounding.(4)Bisulfite sequencing showed that the both CCGG sites of ATP synthase F1 subunit 1 and NADH-ubiquinone oxidoreductase were hypomethylated inducing by mechanical wounding. Therefore, the results suggested the DNA methylation may involved in the response of mechanical wounding in epicormic shoots of rubber trees.