沉香属植物基因组DNA提取及SSR反应体系的优化

摘要

The extraction method of A q uilaria ssp. genomic DNA and optimization of SSR-PCR reaction condition for Aquilaria sinensis （Lour.） Gilg were studied. Genomic DNA was isolated using improved CTAB method. Agarose gel electrophoresis and detection results showed that improved CTAB DNA extraction produced DNA was an efficient DNA extraction method, which yielded high-quality DNA with no degradation. It was effectively removed polysaccharide, polyphenols, and successfully inhibited protein and such impurities as salt ion and RNA. These DNA can be used for SSR molecular marker. The reaction system and amplified procedure suitable for A. sinensis were as follow： 20 μ1 amplification reaction system, 2.5 ng/μL template DNA, 1 μmol/L primer, 2.0 mmol/L MgC12, 200 μmol/L dNTPs and 1.2 U Taq DNA polymerase. Nine of Aquilaria ssp.. DNA samples were used to verify the SSR-PCR reaction system, and could produced qualified fragments. The optimal system could provide a favor able basis for further study on genetic of A. sinensis and Aquilaria ssp.. using SSR molecular marker.%以白木香为材料，研究沉香属植物基因组DNA提取方法，并优化SSR-PCR反应体系。通过改良CTAB法提取白木香叶片基因组DNA，经电泳和吸光度检测。优化影响白木香SSR-PCR主要参数．确立适合沉香属植物的SSR-PCR反应体系和扩增条件：在20斗L反应体系中，模板DNA、引物、Mg^2＋、dNTP和TaqDNA聚合酶等5种主要成分的最适浓度分别为2．5ng／μL、1μmol／L、2mmol／L、0．2mmol／L、1．2U：并用9份沉香属植物DNA样品对SSR-PCR反应体系验证．能扩增清晰条带。SSR优化系统的建立为进一步利用SSR分子标记技术进行白木香种群及不同地区沉香属种群遗传多样性研究提供基础．