水稻AFLP指纹图谱的引物选择研究

Studies on the Primers Screening for AFLP Fingerprints of Rice Cultivars

通过对适合制作水稻品种AFLP指纹图谱的引物进行研究 ,初步建立了一套快速有效地制作AFLP指纹的引物选择法 :第 1步 ,选择亲缘关系相近的 3组以上材料 ,保证组内材料遗传差异较小 ,组间有一定差异 ;第 2步 ,利用两端相同的 16对 2 +2引物组合选择扩增 ,初步选出具有一定多态性的引物 ;第 3步 ,对具有一定多态性的引物重新组合 ,构成两端不同的 2 +2引物组合 ,选出多态性较高的引物组合 ;第 4步 ,将多态性较高的 2 +2引物的一端增加 1个选择碱基 ,构成 2 +3或 3+2引物组合 ,筛选多态性更强、谱带较多、质量较好的引物组合 ;第 5步 ,随机选择部分品种验证所选引物的有效性。选出了M2 1P87和M73 P17等多态性高、谱带多、带纹清晰的引物组合 。

AFLP (amplified fragment length polymorphism) is a very powerful fingerprinting technology. The key of making variety fingerprints is to specially select powerful primers for each crop. A fast and effective procedure of selecting AFLP primers for rice variety fingerprinting was set up:(1) choosing 3 or more group materials that have close genetic relations within groups; (2) selecting potential polymorphic 2+2 primers with same ends; (3) recombining the selected potential polymorphic primers and choosing more polymorphic primers; (4) adding one selecting base at one end to become 2+3 or 3+2 primers and further selecting more polymorphic primers. (5) some primers were selected with this procedure, such as M 21 P 87 and M 73 P 17 , with which the fingerprints had more polymorphism and high quality.