湘产百合核心种质库的SRAP体系的建立

目的确立适用于湘产百合核心种质库的相关序列扩增多态性（SRAP）反应体系。方法根据构建核心种质库的需求,对实验步骤、评分规则进行针对性改良,通过Mg2＋、d NTPs、引物、Taq酶、模板5因素4水平的正交试验与单因素试验构建SRAP体系,并筛选出适用引物。结果最佳反应体系（20μL）：Mg^2＋浓度2.5 mmol/L,d NTPs浓度0.20μmol/L,引物浓度0.20μmol/L,Taq酶1.0 U,模板DNA 50 ng能获得明亮清晰的扩增条带;并由144对引物组合中筛选出32对产物稳定、多态性丰富的引物组合。结论该体系具有较高的稳定性,并可提高引物使用率,满足百合核心种质构建对大信息量的需求,适用于湘产百合核心种质库的研究。

SRAP-PCR System Optimization for Core Collection Establishment of Lily in Hunan

Objective The SRAP system of lily from Hunan province for core collection was established. Methods According to the requirements of building core collection, the experimental procedures and scoring rules were modified. The SRAP system was established through the single factor test and orthogonal design at four levels and five factors： Mg2＋, d NTPs, DNA template, primers and DNA polymerase, and the optimized primers were screened. Results The best reaction system was 20 μL reaction solution, containing 2.5 mmol/L Mg^2＋, 0.20 μmol/L d NTPs, 0.20 μmol/L primers,1.0 U DNA polymerase, 50 ng DNA template. This system showed bright and clear amplified bands.32 primers with rich polymorphism and steady bands were selected from 144 primers. Conclusion This system with high stability could improve the utilization rate of primer and meet the large information for the core collection establishment. The system is applicable to establish the core collection of lily from Hunan province.