“循化红”线辣椒SRAP-PCR反应体系的优化与建立

Optimization and Establishment for SRAP-PCR System of Line Pepper "Xunhuahong"

[目的]建立适合于"循化红"线辣椒基因组的SRAP-PCR优化扩增反应体系。[方法]采用L16(45)正交试验设计,对"循化红"线辣椒SRAP反应体系中的Mg2+浓度、Taq聚合酶、dNTPs浓度、引物浓度、模板DNA浓度进行5因素4水平正交优化。[结果]"循化红"线辣椒的最佳SRAP-PCR反应体系为:Mg2+浓度为2.0 mmol/L,dNTPs为0.5 mmol/L,Taq酶0.5 U,引物浓度为0.4μmol/L,模板DNA浓度为1.0 ng/μl,总体积20μl。[结论]该体系的建立为SARP标记技术应用于"循化红"线辣椒种质资源的收集与利用,品种的鉴定、标记辅助育种等方面奠定了良好的试验基础。

[ Objective ] The research aimed to establish the suitable SRAP-PCR system of line pepper ＂Xunhuahong＂. [ Method ] Orthogonal design was used to optimize SRAP-PCR amplification system of line pepper on four levels of five factors （ Taq DNA polymerase, Mg^2 ＋ , DNA template, dNTPs, and primer respectively）. [ Result] A most suitable SRAP-PCR system for line pepper was established. And the total reaction system was 20 μl contained 0.5 U DNA polymerase, 2.0 mmol/L Mg^2＋ , 1.0 ng/μl DNA template, 0.5 mmol/L dNTPs, and 0. 4 μmol/L primer. [ Conclusion] This optimized system for SRAP marker could provide good basis for further research on germplasm resources collection and utilization, variety identification and marker-assisted selection in line pepper.