海南雷公笋的SCoT-PCR体系优化及有效引物筛选  

Optimization of SCoT-PCR System and Screening of Effective Primers for Costus speciosus of Hainan

作　　者：孙秀秀 王春梅[1,2] 陈加利 夏腾飞[1,2] 冯依欣 郑道君 SUN Xiu-xiu;WANG Chun-mei;CHEN Jia-li(Sanya Institute,Hainan Academy of Agricultural Sciences,Sanya,Hainan 572025;The Key Laboratory of Tropic Special Economic Plant Innovation and Utilization/National Germplasm Resource Chengmai Observation and Experiment Station/Institute of Tropical Horticulture Research,Hainan Academy of Agricultural Sciences,Haikou,Hainan 571100)

机构地区：[1]海南省农业科学院三亚研究院,海南三亚572025 [2]海南省农业科学院热带园艺研究所/国家种质资源澄迈观测实验站/海南省农业科学院海南省热带特种经济植物种质资源创新利用重点实验室,海南海口571100

出　　处：《安徽农业科学》2024年第8期95-99,共5页Journal of Anhui Agricultural Sciences

基　　金：海南省科技专项资助(ZDYF2021XDNY127)。

摘　　要：[目的]分析DNA、引物,dNTPs、Taq DNA聚合酶4种因素对雷公笋SCoT-PCR扩增结果的影响,通过最优的SCoT-PCR反应体系筛选多态性引物,为雷公笋种质资源分子标记研究提供条件。[方法]以海南雷公笋为材料,采用单因素试验和正交试验方法。[结果]Taq酶对雷公笋SCoT-PCR扩增的影响最大,其次是模板DNA和dNTPs,最后是引物;最优反应体系为20μL体系中40 ng DNA,0.30μmol/L引物,0.25 mmol/L dNTPs,3.00 U Taq酶。经验证,该体系获得的扩增产物清晰稳定;应用该体系从40条SCoT引物筛选出8条多态性好,且适合雷公笋扩增的引物,多态性条带占62.64%。[结论]该研究为使用SCoT分子标记技术对雷公笋开展深入研究提供了重要的理论基础和技术支持。[Objective]In order to analyze the effects of DNA,primers,dNTPs and Taq DNA polymerase on the SCoT-PCR amplification results,the polymorphic primers were selected through the optimal SCoT-PCR reaction system to provide conditions for the study of molecular markers of the Costus speciosus resources.[Method]Unifactor test and orthogonal test methods were used.[Result]Taq had the greatest effect on SCoT-PCR amplification,followed by template DNA and dNTPs,and the last was primer.The optimal reaction system was 40 ng DNA,0.30μmol/L primer,0.25 mmol/L dNTPs,3.00 U Taq enzyme in the 20 system.The amplification products obtained by this system were clear and stable.8 primers with good polymorphism were selected from 40 SCoT primers,and the polymorphism bands accounted for 62.64%.[Conclusion]This study provided an important theoretical basis and technical support for further study of Costus speciosus with SCoT molecular marker technology.

关 键 词：雷公笋 SCoT 反应体系优化 有效性引物 

分 类 号：S647[农业科学—蔬菜学]