摘要
以星星草种子在愈伤组织诱导培养基上萌发的纤细幼苗为亲本对照,应用MSAP分子标记技术对其体细胞无性系表观遗传变异进行研究。研究结果表明,16对MSAP引物组合对星星草亲本对照和随机选出的与亲本起源于同一粒种子的3瓶愈伤组织、8株再生苗基因组DNA样品选择性扩增反应,分别检测到163-170个甲基化的5′-CCGG-3′位点;与亲本对照相比较,3瓶愈伤组织中所检测到的基因组5′-CCGG-3′位点上其外侧C半甲基化和内侧C完全甲基化水平均呈轻微地降低趋势;而在其再生苗基因组,上述两种DNA甲基化水平与亲本对照之间差异均不显著;进一步分析相关5′-CCGG-3′位点C甲基化变异模式,表明其体细胞无性系DNA甲基化变异主要发生在内侧C上,且在星星草愈伤组织中内侧C去甲基化变异略多于超甲基化。聚类分析结果表明,基于MSAP数据分析的星星草体细胞无性系样本彼此间平均遗传相似性系数为0.989。
Using the slender shoot that grew up from a seed of Puccinelliatenuifloraon the callus-induction medium as control,the epigenetic variation of clonal lines of somatic cell of Puccinellia tenuiflora was studied with MSAP molecular marker technique.The results showed that 163 to 170 methylated CCGG sites were amplified in the analyzed samples by MSAP using the selected 16 primer combinations.Compared with the control,the levels of hemi-methylation at external C and full-methylation at internal C at CCGG sites revealed in calli were slightly lower.Nevertheless,the level of DNA methylation detected in the regenerated seedlings showed no significant difference with the control.By comparison of the cytosine methylation patterns in a locus-specific manner,it was found that variations mainly appeared at the inner Cs of some CCGG sites,and demethylation occurred a bit more often than hypermethylation at the internal Cs in the 3callus-pools.Cluster analyses indicated that the average genetic similarity values calculated on the basis of d MSAP data among the 8regenerated plants,three pools of calli and the control was 0.989.
出处
《中国草地学报》
CSCD
北大核心
2016年第2期26-33,共8页
Chinese Journal of Grassland
基金
吉林省自然科学基金项目(201115155)
