小麦单片叶片DNA 提取及SSR－PCR

摘要

[目的]使用PCR扩增鉴定哪些微卫星引物可用于鉴定小麦遗传背景下的黑麦遗传成分。[方法]以小麦单片叶片为材料，采用CTAB法提取DNA，以获得的DNA为模板进行SSR－PCR扩增，再进行普通小麦与黑麦之间多态性引物筛选，筛选出可用于鉴定小麦遗传背景下的黑麦遗传成分的引物。[结果]在所研究的20对SSR引物中，有18对在普通小麦与黑麦之间扩增出的产物有差异，引物发生变化的比例为90％。这些产生差异带的多态性引物大体可划分为两大类：一类是在黑麦与普通小麦之间均出现一种或者多种具有差异的产物带，如SCM2、SCM109、SCM180、SCM304，另一类是在黑麦上能扩增但在普通小麦上却未能扩增或者扩增的条带不清晰，如SCM101、SCM120、SCM138、SCM268。[结论]以上这2种情况下的引物均可用于鉴定小麦遗传条件下的黑麦遗传成分。%Objective] To find the micro-satellite primers obtained by PCR amplification, which was suitable for the identification of rye genetic component under wheat genetic background.[Method] With wheat single leaf as the test material, DNA was extracted by CTAB method.SSR-PCR amplification was carried out with the obtained DNA as the template .Screening of pleomorphic primer was carried out between common wheat and rye, so as to find the primer of rye genetic component suitable for the identification of wheat genetic background.[Result] From 20 pairs SSR primers, 18 pairs of SSR primers could amplify the different sites between common wheat and rye .The percentage of primer change was 90%.These polymorphic primers could be divided into two types .There were one or more product bands with differences between common wheat and rye, such as SCM2, SCM109, SCM180 and SCM304.The other type could amplify in rye but could not amplify in common wheat or the band was unclear, including SCM 101, SCM120, SCM138 and SCM268.[Conclusion] Under the above two cases, primers can be used to authenticate genetic constitution of rye under common wheat .