核桃MSAP试验体系建立及引物筛选

Establishment of walnut MSAP reaction system and primers screening

以‘中宁强’核桃品种成熟叶片为材料,通过优化可影响甲基化敏感扩增多态性(Methylation-sensitive amplification polymorphism,MSAP)分析的因素,建立适用于核桃MSAP分析的试验体系,并用于MSAP试验引物筛选。结果表明:采用该优化的MSAP试验体系获得了清晰、重复性好的银染图谱,从90对MSAP引物中,筛选出43对条带清晰且稳定多态的引物,表明该体系可用于核桃DNA甲基化模式分析。

The mature leaves of ‘ZhongNingQiang＇ walnut cultivar were taken for research ma- terials to establish the experimental system for MSAP （methylation-sensitive amplification pol- ymorphism） analysis by optimization of the factors affecting the MSAP system, and primers were screened by this modified MSAP experimental system. The legible and repeatable finger- prints of silver stain were obtained, and 43 legible, stable and polymorphic primer combina- tions were screened from 90 primer combinations, which shows that this modified MSAP sys- tem can be used for DNA methylation pattern analysis of walnut.