菠菜AFLP反应体系初探  

Optimization and Establishment of AFLP Analying System in Spinach (Spinacia oleracea L.)

作　　者：梅燚[1,2] 侯雷平[1] 崔艳玲[2] 陈海丽[2] 孟淑春[2] 

机构地区：[1]山西农业大学园艺学院,山西太谷030801 [2]北京市农林科学院蔬菜研究中心,北京100097

出　　处：《华北农学报》2010年第4期111-115,共5页Acta Agriculturae Boreali-Sinica

基　　金：北京市科技新星计划项目(2008B37);国家科技支撑计划项目(2007BAD85B02;2008BADA6B03);北京市人事局留学人员科技活动市级重点资助项目(20080006);北京市政府购买科技服务项目(20080901)

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20μL,Mg2+(25mmol/L)1.2μL,dNTPs(2.5mmol/L)1.6μL,Taq-polymerase(5U/μL)0.2μL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。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,and 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-pol-ymerase（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.

关 键 词：菠菜 AFLP 影响因子 

分 类 号：S636.1[农业科学—蔬菜学]