盐胁迫下红花基因组DNA甲基化的MSAP分析

以红花幼苗为研究材料,经200mmol.L-1 NaCl溶液分别处理4、8、12h,以未经盐胁迫处理为对照(CK),对DNA甲基化变化特征进行MSAP(甲基化敏感扩增多态性)分析。选用13对选扩引物,共检测到3 140个基因位点。甲基化水平分析表明,CK,4、8、12h处理样本中总甲基化率分别为31.8%、27.1%、31.0%和31.3%。进一步对不同处理时间红花基因组DNA甲基化模式的变化特征进行分析表明,NaCl处理4h后DNA的去甲基化模式调整占优势,而处理8h及12h后DNA甲基化水平恢复到CK水平,甲基化及去甲基化模式调整基本一致。上述结果显示,盐胁迫条件下红花基因组DNA甲基化状态发生了一定变化,并且这种变化与盐胁迫程度相关,推测DNA甲基化修饰可能参与了红花的盐胁迫应答。

Analysis of DNA Cytosine Methylation under Salt Stress in Carthamus tinctorius Based on MSAP

Analysis of DNA methylation status was carried out by MSAP(methylation-sensitive amplified polymorphism) method in Carthamus tinctorius L.after 200 mmol·L-1 NaCl stress for 0,4,8 and 12 h,respectively.Totally,3 140 genetic loci were produced by 13 pairs of selective primers.The results indicated that the corresponding total DNA methylation ratios were 31.8%,27.1% 31.0% and 31.3% in the samples treated by NaCl stress for 0,4,8 and 12 h,respectively.Further analysis of the DNA methylation patterns suggested that demethylation adjustment was predominant in 4 h samples,however,the methylation level was got back to the level of CK in 8 and 12 h samples,and methylation adjustment was almost consistent with demethylation adjustment.These results indicated that the changes of DNA methylation status were closely associated with the responses of salt stress in Carthamus tinctorius L.