扁桃基因组DNA的制备和AFLP技术体系的建立

DNA Template Preparation and the Establishment of AFLP Technique for Almond Cultivar

AFLP技术对DNA模板的质量要求较高。桃属果树扁桃的叶片含有较高的糖分、多酚类等物质,基因组DNA的纯化比较繁琐。笔者采用3×CTAB缓冲液提取基因组DNA,并加以纯化,使DNA质量达到AFLP技术的要求。在AFLP分析中,筛选出合适的引物组合:采用E+3/M+3选择性引物组合时,扩增条带少;改用E+2/M+3引物组合后,扩增产物主要分布在600～100bp内,且扩增条带数目适中,多态性适宜,电泳图谱清晰。

AFLP analysis needs high quality DNA templates.Almond leaves contain high amounts of polyphenolics and polysaccharides,and therefore,the extraction of genomic DNA has been very difficult.In this study,gemomic DNA was extracted from different cultivar of almond using 3×CTAB extraction buffer and then purified with phenol/chloroform.Restriction analysis and preamplification indicated the DNA samples were high quality and therefore,used in the AFLP analysis.Results showed that the E+3/M+3 primer combination produced very few bands on genomic DNA of different almond cultivar,not suitable for the AFLP analysis,and the E+3/M+3 a great number of polymorphic bands in the range of 100～600 bp.The establishment of AFLP technique in this work is significant for the analysis of the genetic relatedness of almond species.