MSAP在水稻害虫白背飞虱中的应用研究

甲基化敏感的扩增多态性分析方法 (MSAP)是在扩增片段长度多态性技术基础上建立起来的主要用来检测样品基因组DNA甲基化情况的一种方法,特别适用于全基因组范围内检测CCGG位点的胞嘧啶甲基化情况,在高等动物和许多植物上已广泛应用,但尚未有在昆虫中应用该方法的报道。从DNA提取、酶切基因组DNA用量、酶切和接头连接时间、电泳图谱显影方法等方面对MSAP方法进行了改进,并在水稻重要害虫白背飞虱上得到成功应用。这为将来在其它昆虫中使用该方法研究基因组DNA甲基化情况提供了一定借鉴和指导价值。且通过比较白背飞虱雌雄性别间和长短翅型间的MSAP图谱,发现雌雄性别间和长短翅型间的基因组DNA甲基化式样都存在明显的差异,说明DNA甲基化可能参与了性别和翅型分化的调节。

The Application of MSAP in Sogatella furcifera (Horvath)

The methylation sensitive amplification polymorphism (MSAP) technique is an amplified fragment length polymorphism (AFLP) based method for detection of DNA methylation. It represents a reliable technique for the detection of cytosine methylation alterations at the site of CCGG in a genome wide perspective. MSAP has been extensively applied in plants and some animals, but no application in insects has been reported yet. The method was improved from DNA extraction, the amount of genomic DNA used for restriction enzyme digestion, time for enzyme digestions and ligation of adapters, and electrophoresis imaging method. And the method was applied successfully in Sogatella furcifera (Horvath), an important rice pest in China. It will provide some references and guiding values for the study of genome DNA methylation in other insects by using this method in the future. By comparing MSAP maps between male and female or between long wing and short wing of S. furcifera, it was found that there were obvious differences in DNA methylation patterns. This indicates that DNA methylation may be involved in gender and wings type differentiation.