火龙果茎基因组DNA提取方法改良  

Improvement in Extraction Methods of Genomic DNA from Pitaya Stems

作　　者：农全东 张明永[1,2] 张美[2] 简曙光[2] 陆宏芳[2] 夏快飞[1,2] 文和明 Nong Quandong;Zhang Mingyong;Zhang Mei;Jian Shuguang;Lu Hongfang;Xia Kuaifei;Wen Heming(Key Laboratory of South China Agricultural Plant Molecular Analysis and Genetic Improvement, South China Botanical Garden, CAS, Guangzhou 510650, China;Guangdong Provincial Key Laboratory of Applied Botany, South China Botanical Garden, CAS, Guangzhou 510650, China;Wenshan Academy of Agricultural Sciences, Wenshan 663099, China)

机构地区：[1]中国科学院华南植物园中国科学院华南农业植物分子分析与遗传改良重点实验室,广州510650 [2]中国科学院华南植物园广东省应用植物学重点实验室,广州510650 [3]文山州农业科学院,文山663099

出　　处：《植物学报》2019年第3期371-377,共7页Chinese Bulletin of Botany

基　　金：中国科学院A类战略性先导科技专项(No.XDA13020500);广州市科技计划(No.201804010409)

摘　　要：火龙果(Hylocereus undulatus)是近年发展起来的一种新兴热带水果,其茎富含多糖、多酚及其它次生代谢物,黏性极大,很难从中提取高质量的DNA。特别是一年生以上的老茎,目前尚未有较好的DNA提取方法。为了解决这一难题,该研究对CTAB+Tris-HCl洗涤法进行了3种方式的改良。结果表明,"改进三"方法可不受取样时期和取样部位的限制,从一年生以上火龙果茎中提取的DNA质量最好且不含黏性物质,可用于酶切与分子标记等生化和分子生物学实验。该研究探索了一条较为理想的火龙果茎DNA提取方法,值得推广应用。Pitaya (Hylocereus undulatus) is a tropical fruit developed in recent years. Because its stem is full of polysaccharides, polyphenols and other secondary metabolites, extracting high-quality DNA from the stem is difficult, especially from stems more than 1 year old. At present, there is no good method for DNA extraction. We examined three methods to improve DNA extraction based on the CTAB+Tris-HCl washing method for better and easier DNA extraction. The third method obtained the best-quality DNA without sticky substances from pitaya stem more than 1 year old and was not restricted by sampling data and sampling site. The extracted DNA can be used in enzyme digestion and inter-simple sequence repeat amplification experiments. Our study provides an ideal method for extracting DNA from pitaya stem, which is worth popularizing.

关 键 词：火龙果 茎 DNA提取 改进CTAB+Tris-HCl洗涤法 

分 类 号：S667.9[农业科学—果树学]