橡胶树MSAP反应体系的建立及其对无性系DNA的甲基化分析

Establishment of MSAP analysis system for rubber tree and its preliminary application in juvenile-type clone

采用改良CTAB法提取橡胶树幼嫩叶片的基因组DNA,通过电泳检测酶切与连接、预扩增和选择性扩增等,建立橡胶树MSAP甲基化敏感扩增多态性(MSAP)分析体系,并利用MSAP技术在基因组水平上对橡胶树幼态优势的形成机理进行研究。结果表明:利用该体系可获得扩增位点多、条带清晰、重复性好的图谱(共扩增出740个条带,404个甲基化位点,平均多态性比例为54.6%),并能有效区分基因组DNA甲基化的状态,为开展橡胶树同一品种不同类型种植材料的表观遗传变异研究奠定了基础。

The genomic DNA from the fresh leaves of rubber tree was extracted with improved CTAB method,and then a MSAP analysis system suitable for rubber tree was established by detecting results of digestion and ligation,pre-amplification and selective amplification key procedure in gel electrophoresis,in order to study formation mechanism of rubber tree young state dominance at the genome level by means of this MSAP technique.The results showed that this system could get many amplified loci,clear band,and good reproducibility map（740 bands,404 methylaed sites,54.6% polymorphic ratio on average）,and status of genomic DNA methylation was distinguished effectively,which suggested that the reaction system was established successfully and can be taken as a base for future epigenetic change study of different type of planting materials from the same rubber tree clone.