正交设计优化花菜类SRAP分子标记体系的研究  

Optimization of the SRAP reaction system in broccoli and cauliflower by orthogonal design

作　　者：李媛[1,2] 姚雪琴[1] 谢祝捷[1] 龚义勤[2] 柳李旺[2] 

机构地区：[1]上海市农业科学院园艺研究所上海市设施园艺技术重点实验室,上海201106 [2]南京农业大学园艺学院,南京210095

出　　处：《上海农业学报》2010年第1期42-45,共4页Acta Agriculturae Shanghai

基　　金：上海市科委科技攻关计划(07DZ19602;08DZ1906304)资助

摘　　要：采用正交设计,从Mg2+、dNTPs、引物三种因素3个水平对青花菜SRAP反应体系进行优化,确立适合青花菜的SRAP反应体系,并在6个青花菜和花椰菜品种中进行验证。结果表明:20μL反应体系中适宜体系的浓度dNTPs为0.2mmol/L、Mg2+为1.25mmol/L、上下引物各0.4μmol/L、DNA为20ng、Taq酶1.2U、退火温度为50℃。By an orthogonal design the reaction system of brocco＇s SRAP（sequence-related ampli- fied polymorphism）was optimized in terms of Mg2＋ ,dNTPs and primers three factors and their three levels,and then the optimized SRAP reaction system was verified with 6 broccoli and cauliflower varieties. The result indicated that in a 20 μL reaction system the appropriate concentrations of dNTPs, Mg2＋ ,and forward and reverse primers were respectively 0.2 mmol/L, 1.25 mmol/L,0.4 μmol/L and 0.4 μmol/L,the DNA of template was 20 rig,and Taq DNA polymerase 1.2 U,the best annealing temperature was 50℃. The results offer the technique basis of using SRAP technology to study broccoli germplasm resource.

关 键 词：青花菜 花椰菜 SRAP 体系优化 

分 类 号：S635.3[农业科学—蔬菜学]