苦瓜外观性状AFLP反应体系的建立及优化

Establishment and Optimization of AFLP Markers in Momordica charantia L.

为探讨苦瓜性状遗传多样性奠定基础,通过对酶切方法的比较研究,建立并优化了苦瓜AFLP反应体系,采用TaqI/VspI或EcorI/MseI2种限制性内切酶酶切组合对的苦瓜(Momordica charantia.L.)的遗传多样性进行研究,探索适合其AFLP的双酶切体系。结果表明EcorI/MseI内切酶进行双酶切时酶切效果较好并且扩增出的条带带纹清晰,扩增信号强度基本一致符合研究需求,并初步筛选得到了32对清晰、多态性高的AFLP引物。说明最适宜的酶切方法是使用EcorI/MseI组合进行双酶切,通过重复试验证明了该体系稳定可靠,可用于苦瓜性状遗传多样性的分析。

For the study were provided the basis the genetic diversity, by companng different enzyme cut method, set up and optimize the balsam pear AFLP reaction system, two kinds of AFLP reaction system with the different restriction enzyme combinations （TaqIⅠ/VspⅠ and EcorⅠ/Msel） were compared charantia L.. The results showed that the effect of EcorⅠ/MseⅠ enzymes was best one, polymorphic bands were clear and the augmentation signal strength were basically the same, 32 primers were screened preliminary with stable amplification and rich polymorphism. The most appropriate enzymes that cut method was the use of EcorⅠ/MseⅠ combination for double enzyme cut, through the repeat tests showed that this system was stable and reliable, and can be used in the genetic diversity of Momordica eharantia L.