适合MSAP分析的核桃子叶DNA提取方法研究.pdfVIP

07 适合MSAP 分析的核桃子叶DNA 提取方法研究 周贝贝，陈凌娜，王伟，马庆国，宋晓波，张俊佩，裴东* 中国林业科学研究院林研所，北京 100091 摘要：核桃成熟子叶组织脂肪酸含量较高，组织培养后多糖多酚含量升高，而 DNA 中多糖 等杂质的残留对酶切等影响较大。为获得适合 MSAP 分析的高质量 DNA ，本研究以‘辽宁 1 号’核桃品种成熟子叶和经过组织培养的近成熟子叶为材料，以叶片为对照，用 3 种方法 （SDS、高盐 CTAB、改良高盐 CTAB ）提取DNA ，对提取的DNA 质量采用琼脂糖凝胶电 泳和分光光度计法进行检测。结果表明：除 SDS 方法外，高盐 CTAB、改良高盐 CTAB 均 能提取到较高质量的叶片 DNA 。SDS 方法提取未能提取出成熟子叶和组培子叶的 DNA ；高 盐 CTAB 提取出较高质量成熟子叶 DNA ，但量较少，未能提取出组培子叶 DNA ；改良 CTAB 提取出较高质量的成熟子叶和组培子叶 DNA ，含量较高，适合进行MSAP 分析。本研究发 现适合 MSAP 分析的核桃成熟子叶 DNA 提取方法，为下游分子生物学实验奠定基础，同时 为其它木本植物 DNA 提取过程中多糖去除的研究提供参考。 关键词：核桃；子叶 DNA 提取；多糖；MSAP The DNA extraction method of walnut cotyledon suitable for MSAP analysis Zhoubeibei，Chenlingna，Wangwei ，Maqingguo，Songxiaobo，Zhangjunpei ， Peidong （Chinese Academy of Forestry，Beijing 100091 ） Abstract: It is high for fatty acid content in walnut mature cotyledon tissue, with polysaccharide polyphenol content increase after tissue culture, and there are great influences to enzyme for DNA polysaccharides residual, etc. To obtain high quality DNA suitable for the analysis of the MSAP, the Liaoning1 walnut varietie’s mature cotyledon and tissue cultured cotyledon were adopted as research material for DNA extraction analysis on 3 kinds of DNA extraction method (SDS and high salt CTAB and modified high salt CTAB) , with leaf tissue as control, and the extracted DNA quality were detected using agarose gel electrophoresis and spectrophotometer method. It conducted that the high salt CTAB and modified high salt CTAB method, except for SDS method, can extract DNA with high quality. The SDS method failed to extract the mature cotyledon and cultured cotyledon’s DNA; the high salt CTAB method extracted the mature cotyledon’s DNA, with high quality but li