草莓属植物SCoT分析体系的建立及优化  

Establishment and optimization of SCoT-PCR system in Fragaria

作　　者：秦国新[1,2] 何桥[1] 梁国鲁[1] 雷家军[3] 郭启高[1] 

机构地区：[1]西南大学园艺园林学院,重庆北碚400715 [2]山西农业大学园艺学院,山西太谷030801 [3]沈阳农业大学园艺学院,沈阳110161

出　　处：《果树学报》2012年第3期393-397,共5页Journal of Fruit Science

基　　金：国家自然科学基金(30971976)

摘　　要：为利用SCoT技术开展草莓属(Fragaria)植物种质资源遗传多样性及分子系统学研究,以草莓属植物11个野生种和1个栽培品种为试材,采用L25(56)正交试验设计,建立了草莓属植物的最优SCoT-PCR反应体系,即20μL反应体系中,含Mg2+2.0 mmol·L-1、dNTPs 0.3 mmol·L-1、Taq酶0.75 U、引物0.75μmol·L-1、模板DNA 40 ng,并对退火温度进行了优化,对电泳检测方法及草莓属植物DNA提取方法进行了改进。共筛选出22个适于进行草莓属植物SCoT分析的引物。经验证,该反应体系稳定性好、可重复性强、多态性高、扩增效果好,适于草莓属植物的SCoT分析。Start Codon Targeted Polymorphism （SCOT） is a novel gene-targeted molecular marker tech- nique. In this experiment, a L25 （5^6） or thogonal design was used for the optimal SCoT-PCR reaction system of strawberry with 11 Fragaria species and a cultivar as materials. The optimal PCR reaction mixture of SCoT-PCR system for strawberry was 2.0 mmol·L^-1 Mg^2＋,0.3 mmol·L^-1 dNTPs,0.75 U Taq polymerase, 0.75 μmol·L^-1 primer and 40 ng template DNA in 20 μL mixtures. Furthermore, the annealing temperature of each primer was optimized; the methods of electrophoresis and strawberry DNA extracting were improved. 22 primers were selected which were suitable to the SCoT-PCR reactions. The optimal reaction system with high polymorphism, stability and repeatability was obtained.

关 键 词：草莓属 SCoT 体系优化 

分 类 号：S668.4[农业科学—果树学]