为分析竹子年龄变化与基因组DNA甲基化之间的相关性, 以5年、31年和>60年起源(从种子萌发年龄算起)的毛竹当年生叶片为材料, 采用35对引物对其进行MSAP检测。结果表明: 3个年龄段的总甲基化率和全甲基化率分别为24.44%、28.21%、32.12%和16.57%、19.41%、21.23%; 发生DNA甲基化的变异位点为52.3%, 去甲基化变异位点为10.3%。可以看出, 随着年龄的增加, 毛竹基因组DNA甲基化敏感多态性呈上升趋势。总甲基化率单因素方差分析的结果表明相同年龄的毛竹个体间没有差异(P=0.307 > 0.05), 而不同年龄间的差异达极显著水平(P＜0.001)。同时, 对所用引物组合进行分析后发现有6对引物(E3/HM2、E3/HM6、E3/HM7、E4/HM5、E4/HM6和E5/HM5)扩增出的位点与总趋势显著相关, 为进一步开展深入研究奠定基础。
In order to probe the features of DNA methylation for bamboo stand with different chronological ages, the technique of methylation-sensitive amplified polymorphism (MSAP) was employed to detect DNA methylation in the paper. Experiment material is Moso bamboo (Phyllostachys heterocycla var. pubescens) leaves with 3 various chronological ages (5, 31, and >60 years after seed germination). During the procedure of genome DNA extration and MSAP analysis, total 35 pairs of MSAP primers were amplyfied. The results showed that MSAP value for bamboo with those three chrono-logical ages were respectively 24.44%, 28.21% and 32.12%, and full-methylation ratios were 16.57%, 19.41% and 21.23%. Meanwhile, the value of variable sites for methylation reached 52.3% and for demethylation was 10.3%. Therefore, it could be concluded that with ages increasing MSAP value rising for Moso bamboo. Moreover the result of variance analysis for methylation ratio indicated that no significant (P=0.307 > 0.05) difference among individuals with the same ages, while significant (P < 0.001) difference exsited among different chronological ages. Throuygh ANOVA it showed that 6 pairs (E3/HM2, E3/HM6, E3/HM7, E4/HM5, E4/HM6and E5/HM5) of primers had obvious influence on DNA methylation for ones with different chronological ages and could be used for further research.
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