整粒和半粒青稞种子DNA的提取及RAPD检测  

DNA Extraction and RAPD Detection of Half-grain and Whole-grain Hordeum vulgare var. nudum

作　　者：郝豆豆 张勇群[1] 高玉花[2] 雷鸣[2] 武俊喜[3] 拉多[2] HAO Dou-dou ZHANG Yong-qun GAO Yu-hua LEI Ming WU Jun-xi LHA-duo(Hospital of Chengdu Office of People＇ s Government of Tibetan Autonomous Region, Sichuan Chcngdu 610000, China Tibet Univer- sity, Tibet Lhasa 850000, China Institute of Geographical Sciences, Tibet Lhasa 850000, China)

机构地区：[1]西藏自治区人民政府驻成都办事处医院,四川成都610000 [2]西藏大学,西藏拉萨850000 [3]中国科学院地理科学与资源研究所,西藏拉萨850000

出　　处：《西南农业学报》2017年第6期1257-1261,共5页Southwest China Journal of Agricultural Sciences

基　　金：湖南师范大学蛋白质与发育生物学教育部重点实验室(2014年)开放课题"几种濒危藏药材的DNA条形码研究"(2015DF03);西藏自治区科技厅自然科学基金项目"濒危藏药材的生物信息学研究"(2015ZR-13-5)

摘　　要：【目的】从青稞种子中提取高质量的基因组DNA。【方法】以半粒无胚青稞种子和整粒青稞种子为材料,用6种不同的方法提取基因组DNA,用琼脂糖凝胶电泳检测DNA的完整性,紫外分光光度计法检测DNA的浓度和纯度,并对提取的DNA进行了RAPD-PCR扩增检测,同时将剩下的半粒有胚青稞种子进行萌发实验。【结果】从半粒和整粒青稞种子中都可以提取到高质量的DNA,用提取的DNA作为RAPD-PCR扩增检测模板,得到的扩增产物条带清晰,多态性条带丰富,其质量满足RAPD分子标记的要求。【结论】高盐低p H法提取的DNA质量最好,是这6种方法中的最佳方法;半粒青稞有胚种子也能够正常生根发芽,可作为遗传实验的材料。[ Objective] The high-quality DNA could be extracted from Hordeum vulgare var. nudum. [ Method] The genomic DNA was ex- tracted by six different methods from half-grain embryoless seeds and whole-grain seeds of Hordeum vulgare var. nudum, UV Spectrophotom- etry and agarose gel electrophoresis were used to detect the concentration, purity and completeness of DNA. RAPD-PCR amplification was carried out on the extracted DNA. At the same time, the left half-grain embryo seeds were cultured for germination experiments. [ Result] High-quality DNA was able to be extracted from both half-grain and whole-grain Hordeum vulgare var. nudum seeds. The extracted DNAs were employed as the template for RAPD-PCR amplification detection, and the products of amplification have clear and rich polymorphism bands, the quality of extracted DNA could meet the requirement of RAPD molecular marker experiment. [ Conclusion] High salt and low pH which could extract high quality DNA is the best method of six methods. In addition, the sprouting experiments indicated that the left half- embryo seed was able to sprout normally and can be used as material of genetic experiment.

关 键 词：青稞种子 基因组DNA RAPD-PCR扩增 

分 类 号：S512.3[农业科学—作物学]