摘要： 提取菠菜嫩叶DNA,运用改良CTAB法提取6份菠菜品种的DNA,对AFLP反应体系的DNA用量、酶切连接、预扩、选扩等试验条件进行了优化分析,初步建立适合于菠菜作物的AFLP反应体系.结果表明:①在PCR仪中酶切的效果比水浴的要好,酶切反应对酶切时间和DNA的浓度要求不敏感.②菠菜AFLP预扩选扩体系的反应体积为20 μL,Mg2+ (2 mmol/L) 1.2 μL,dNTPs (2. mmol/L) 1.6 μL,Taq-polymerase ( U/μL) 0.2 μL最佳.为菠菜AFLP分子标记的品种亲缘关系鉴定和遗传育种等提供一定的理论基础.

关键词: 菠菜, AFLP, 影响因子

Abstract: In this study toal genetic DNA from tender leaves of Spinach (Spinacia oleracea) was extracted with improved CTAB. An AFLP(amplified fragment length polymorphism) analysis system was established by optimizing the concentration of template DNA，DNA restriction reaction，ligase，reaction，pre-amplification and selective amplification. The results were obtained as follows: ①The effect of restriction enzyme was better in PCR instrument than in waterbath， a nd the reaction time and the concentration of DNA were not sensitive to restriction enzyme. ②The best reaction system in 20 μL total volume contained Mg²⁺ (25 mmol /L) 1. 2 μL，dNTPs (2. 5 mmol /L) 1. 6 μL and Taq-polymerase(5 U /μL) 0. 2 μL. This paper provided a method for further study in the field of genetic relative analysis of Spinach (Spinacia oleracea L.) varieties and genetic breeding.

Key words: Spinach (Spinacia oleracea L.), AFLP, Influencing factors

中图分类号: 

• S636.1