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目的:建立适于白木通的SRAP反应体系,为白木通的遗传多样性研究奠定基础。方法:以白木通基因组DNA为模板,优化了SRAP反应体系的各主要参数。结果:建立稳定可靠的SRAP-PCR反应体系(25μL):模板DNA 90 ng,dNTPs 200μmol/L,TaqDNA聚合酶1 U,引物0.4μmol/L,10×PCR Buffer 2.5μL;反应程序中第2次最适退火温度为55℃;筛选出12对稳定性好、多态性高的SRAP引物;并对5个白木通进行扩增验证,共获得114个多态性位点。结论:该体系是适于白木通的SRAP反应体系,为后续白木通遗传多样性研究奠定了基础。
OBJECTIVE: To establish a SRAP reaction system suitable for Baimundong, laying a foundation for the study of genetic diversity of Baimutiaoningin. Methods: The main parameters of SRAP reaction system were optimized by using genomic DNA of Baimu Tong as a template. Results: A stable and reliable SRAP-PCR reaction system (25 μL) was established: 90 ng of template DNA, 200 μmol / L of dNTPs, 1 U of Taq DNA polymerase, 0.4 μmol / L of primer and 2.5 μL of 10 × PCR Buffer. The optimum annealing temperature was 55 ℃. Twelve pairs of SRAP primers with good stability and high polymorphism were screened out. A total of 114 polymorphic loci were obtained from 5 white wood samples. Conclusion: This system is suitable for the SRAP reaction system of Baimu, which lays the foundation for the follow-up study on the genetic diversity of Baimuntong.