摘要
白木香为瑞香科沉香属植物,是中国特有的珍贵药用植物,也是国产沉香药材唯一资源植物,现已濒临灭绝,被载入《中国植物红皮书》。采用改良CTAB法提取白木香DNA较传统方法简单高效。通过琼脂糖凝胶电泳和OD260/OD280比值测定,所得基因组DNA纯度高,可作为ISSR等以PCR为基础的DNA多态性标记。通过优化影响白木香ISSR-PCR的主要参数,确立了适用于白木香的ISSR反应体系和扩增条件。结果表明在20μl反应体中,模板DNA、引物、Mg2+、dNTP和Taq DNA聚合酶5种主要成分的最适浓度分别为25ng、0.8μmol/L、2.5mmol/L、0.2mmo1/L、1.0U,扩增出足量产物至少需要35个循环。
Aquilaria sinensis （Lour.） Gilg classified as Thymelaeaceae, is one of the valuble medicinal plants in China. But its wilding resource has been profoundly damaged, and has been recorded in 《China Plants Red Book》. The improved CTAB method which can extracting high-quality genome DNA of Aquilaria sinensis （Lour.） Spreng was much more simple and efficient than the conventional methods. And the extracted DNA was suitable for the polymorphic analysis based on PCR, such as ISSR-PCR. The optimal ISSR-PCR reaction conditions in A quilaria sinensis （Lour.） Spreng was established by studying the main parameters. Results showed that the optimum concentration of five important components i.e. template DNA, primer, Mg^2＋, dNTP, Taq DNA polymerase in 20ul reaction mixture was 25 ng,0.8umol/L,2.5 mmol/L,0.2 mmol/L, 1.0U, respectively, and at least 35 PCR cycles should be carried out to ensure sufficient PCR products.
出处
《中国农学通报》
CSCD
北大核心
2009年第2期250-254,共5页
Chinese Agricultural Science Bulletin
基金
中央级公益性科研院所基本科研业务费专项资金"重要基因资源发掘与分子育种"(ITBBZD2007-2-2)资助
关键词
白木香
DNA提取
ISSR
PCR反应体系
A quilaria sinensis（Lour.） Spreng, extracting DNA, ISSR, PCR reaction conditions
