SCAR标记转化失败的原因和对策.pdfVIP

分子植物育种，2010 年，第8 卷，第3 期，第589-594 页 Molecular Plant Breeding, 2010, Vol.8, No.3, 589-594 评述与进展 Review Progress SCAR 标记转化失败的原因和对策 1 1 1* 1 2 孙保娟 李植良 黎振兴 罗少波 李艳艳 1 广东省农业科学院蔬菜研究所, 广州, 510640; 2 华南农业大学生命科学院, 广州, 510642 * 通讯作者, lizhxgaas@163.com 摘 要 SCAR 标记作为一种快速、简便可以直接应用的分子标记，在标记辅助选择育种中发挥着重要的作 用。但是目前SCAR 标记的转化效率不是很高。本文从引物设计、序列相似或多拷贝及DNA 甲基化等方面分 析SCAR 标记转化失败的可能原因；同时从引物设计技巧、PCR 反应条件、借助CAPS 和SSCP 技术以及借助 原始标记邻近序列等方面综述了SCAR 标记转化失败的情况下获得序列特异性标记的几种有效补救手段。 关键词 SCAR 标记, SCAR 转化失败, 原因和对策 Reasons for SCAR Conversion Failure and its Remedies Sun Baojuan 1 Li Zhiliang 1 Li Zhenxing 1* Luo Shaobo 1 Li Yanyan 2 1 Vegetable Research Institute, Guangdong Academy of Agricultural Sciences, Guangzhou, 510640; 2 College of Life Sciences, South China Agricultural University, Guangzhou, 510642 * Corresponding author, lizhxgaas@163.com DOI: 10.3969/mpb.008.000589 Abstract SCAR marker, as a fast, simple and direct useable molecular marker, played an important role in mark- er-assisted selection breeding. However, the conversion rate of SCAR markers is not very high at present. In this sum- mary, we analysed the possible causes of SCAR conversion failure from the aspects, such as primer design, mul- ti-copy or similarity of sequence, and DNA methylation, etc. At the same time, when the conventional SCAR marker conversion was failed, several effective remedy approaches for obtaining sequence-specific markers were overviewed from the aspects of primer design, PCR reaction condition, by CAPS and SSCP technology and with the aid of the flanking sequences of the origin markers. Keywords SCAR marker, SCAR conversion failure, Reasons and remedies 序列特异扩增区域(sequence characterized am- 性，为共显性的标记，待检测DNA 间差异可直接通 plified region, SCAR) 标记是 Paran 和 Miehelmore 过有无扩增产物显示，SCAR 标记一般可以通过琼 (1993)提出的，首次在RAPD (random amplified poly- 脂糖凝胶检测，具有快速、简便、成本低廉等优点，适 morphic DNA)标记基础上进行转化并应用。发展该 用于大量样品的快速鉴定和分析。对