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从杂交油葵A1 5及其亲本的 1 / 2粒干种子中提取基因组DNA ,选用 1 7对引物组合进行AFLP分析 ,构建了它们的指纹图谱。 1 7对引物在A系与R系当中共扩增出 1 1 2 5条扩增产物 ,其中 1 4 4条带表现出多态性 ,平均每对引物扩增 66条带 ,不同引物组合产生的DNA片段数目在 50～ 70之间 ,大小分布于 1 0 0bp～50 0bp ,多态性比率为 1 2 .8%。从中筛选出的 2对引物E_AAC/M_CTC和E_ACG/M_CTG可将亲本和子代区分开 :引物对E_AAC/M_CTC在A系中扩增出 440bp、1 90bp、1 60bp 3条特征谱带 ,在R系中扩增出 380bp、350bp、2 2 5bp、1 80bp 4条特征谱带 ,E_ACG/M_CTG在A系中扩增出了 2条特征带 480bp和2 65bp,在R系中扩增出 490bp、2 2 0bp、2 0 5bp、1 2 5bp4条特征谱带 ,且上述谱带均在子代中出现。用引物组合E_ACG/M_CTG对A1 5、双亲以及与A1 5外型十分相似的 1 0个常用油葵杂交种进行AFLP分析 ,不仅表现出良好的多态性 ,并能够清楚地将它们加以区分。以其对 50粒A1 5杂交种子进行纯度鉴定 ,得到与大田纯度检测一致的结果 ,说明使用AFLP标记检测油用向日葵的品种和纯度是可行的。对现行种子纯度和品种鉴定的方法进行了讨论。
Genomic DNA was extracted from 1/2 dry seeds of hybrid oil sunflower A1 5 and its parents, and AFLP analysis was performed on 17 primer combinations to construct their fingerprints. A total of 1125 pairs of primers were amplified in the A and R lines, of which 144 bands showed polymorphism with an average of 66 bands per primer pair, and different combinations of primers produced The number of DNA fragments ranged from 50 to 70 and the size ranged from 100 bp to 50 0 bp, with a ratio of 12.8%. Two pairs of primers, E_AAC / M_CTC and E_ACG / M_CTG, were selected to separate the parents and offspring. The primer pairs E_AAC / M_CTC amplified 440bp, 1 90bp and 160bp three characteristic bands in line A, Four characteristic bands of 380bp, 350bp, 2 2 5bp and 180bp were amplified by PCR. Two characteristic bands of E_ACG / M_CTG were amplified in A line with 480bp and 2 65bp and 490bp in R system 2 0bp, 2 0 5bp and 1 2 5bp bands, all of which appeared in the offspring. AFLP analysis of A1 5, parents, and 10 commonly used oil sunflower hybrids with primer combinations E_ACG / M_CTG not only showed good polymorphism but also clearly distinguished them. The purity of 50 A1 5 hybrid seeds was identified and the result was consistent with the field purity test. It is feasible to detect the varieties and purity of oil sunflower by AFLP. The current methods of seed purity and variety identification are discussed.