关键词: 益智, SRAP标记, 体系优化, 引物筛选

Abstract: In order to establish the optimal SRAP-PCR system for southern Chinese herbs Alpinia oxyphylla Miquel and screen the specific primers for studying the genetic diversity of Alpinia oxyphylla Miquel in different geographical groups, the different geographical groups of Alpinia oxyphylla resources in Hainan were collected, and the genomic DNA of Alpinia oxyphylla Miquel was extracted by plant genome DNA extraction kit and then the purity and concentration of the DNA samples were detected. The optimal reaction system (total volume of 25 μL) was obtained by orthogonal test: Taq DNA polymerase 0.5 U, dNTPs 4 mmol/L, Mg2+ 2 mmol/L, primers each 2 μmol/L, DNA templates 10 ng, 10×PCR buffer 2.5 μL. 7 pairs of specific primers were screened by the optimal reaction system from 144 primers, and the sizes of their amplified fragments were within 100~2 000 bp and evenly distributed. And the ratios of polymorphic bands were above 85%. The results of this study laid a foundation for the study on genetic diversity of southern Chinese herbs Alpinia oxyphylla Miquel.

Key words: Alpinia oxyphylla Miquel, SRAP marker, system optimization, primer screening