野外保存对益智叶片基因组DNA的影响  

Effects of Wild Preserved Methods on Genomic DNA from Alpinia oxyphylla Leaves

作　　者：高炳淼[1] 王朝[1] 何萍[1] 田建平[1] 张俊清[1] 魏娜[1] 

机构地区：[1]海南医学院海南省热带药用植物研究开发重点实验室,海南海口571199

出　　处：《贵州农业科学》2015年第7期19-22,25,共5页Guizhou Agricultural Sciences

基　　金：海南省自然科学基金项目"益智遗传多样性的AFLP分析"(814291);海南医学院药学院大学生创新课题"益智AFLP-PCR体系的建立与优化"(Hyydc2013004);海南医学院引进人才科研启动经费

摘　　要：为获得高质量的益智基因组DNA寻找一种简单可行的野外益智鲜叶保存方法,利用植物基因组提取试剂盒法提取益智基因组DNA,以新鲜叶片为对照及DNA的纯度、浓度、琼脂糖凝胶电泳和ITSPCR扩增效果为评价指标,比较室温阴干、室外晒干、硅胶干燥和液氮保存4种鲜叶保存方法对提取益智基因组DNA效果的影响。结果表明:4种保存方法均可获得益智基因组DNA且ITS-PCR均获得约700bp的PCR产物,其中液氮保存5d的叶片提取DNA条带清晰明亮,室温阴干、硅胶干燥和室外晒干保存方法提取的基因组DNA分别在4d、3d和2d后开始发生降解,不同保存方法效果依次是液氮保存>室温阴干>硅胶保存>室外晒干。在远距离野外资源调查和收集时,采用室温阴干保存新鲜益智叶片是经济简单可行的野外保存方法。In order to find a simple and practical method to save the wild A. oxyphylla fresh leaves, so as to obtain high quality genomic DNA, A. oxyphylla genomic DNA was extracted using the plant genome Extraction Kit. Compared with fresh leaves, four different preserved methods （dried at room temperature, outdoor season, dried with silica gel, stored in liquid nitrogen） for A. oxyphylla leaves were evaluated. The effects of which were compared based on the yield, purity, agarose gel electrophoresis and ITS-PCR amplification effect, to identify genomic DNA of leaves which were saved after different methods. Results： Four kinds of preservation methods can get A. oxyphylla genomic DNA and ITS-PCR were obtained approximately 700 bp PCR product. Genomie DNA was extracted from leaves stored in liquid nitrogen five days, of which bands showed clear and bright, Genomic DNA was extracted from leaves stored in dried at room temperature, silica gel drying and outdoor season, which respectively start degraded after 4 d, 3 d and 2 d. The effect of different preservation methods are stored in liquid nitrogen 〉 dried at room temperature 〉 dried with silica gel 〉 outdoor season. Therefore, fresh A. oxyphylla leaves stored at room temperature drying is both economical and feasible method of field preservation in the distant field resources investigation.

关 键 词：益智 DNA提取 PCR 野外保存方法 

分 类 号：S567.79[农业科学—中草药栽培]