枇杷属植物基因组DNA提取方法的改进及其应用An improved procedure for nuclear DNA isolation from Eriobotrya plants and its application
刘月学,杨向晖,林顺权,胡桂兵,刘成明
摘要(Abstract)：
以枇杷属的普通枇杷和多种野生枇杷的叶片为材料,对枇杷的DNA提取进行了研究。针对枇杷富含多酚类 等次生物质的特点对枇杷DNA的提取步骤进行了改良处理:采用CTAB-蛋白酶K法加重复抽提,去除枇杷材料中 的相关次生物质,获得了高质量的DNA。所提取的DNA完全能满足PCR、RAPD、AFLP等一些分子生物学实验要 求。
关键词(KeyWords)： 枇杷属;DNA提取
基金项目(Foundation): 教育部骨干教师资助计划 广东省农业攻关计划项目(2004B20901001)。
作者(Author): 刘月学,杨向晖,林顺权,胡桂兵,刘成明
Email:
DOI: 10.13925/j.cnki.gsxb.2005.02.023
参考文献(References)：
- [1]林顺权,杨向晖,刘成明,等.中国枇杷属植物的自然地理分布[J]. 园艺学报,2004,31(5):569-573.
- [2]郑国华,郑育俊,何文庆.枇杷基因组DNA分离技术及浓度测定 [J].福建农业科技,2000,(6):13-15.
- [3]程运江,伊华林,庞晓明,等.几种木本果树DNA的有效提取[J]. 华中农业大学学报,2001,10:481-483.
- [4] Badenes ML, Canyamas T, Romero C, et al. Genetic diversity of a European collection of loquat [Eriobotrya japonica(Thunb) Lindl.] determined by molecular markers[C]. Options Mediterraneennes:Proceedings from the first International loquat symposium, 2003.53-56.
- [5]潘新法,孟祥勋,曹广力,等.RAPD在枇杷品种鉴定中的应用[J]. 果树学报,2002,19(2):136-138.
- [6]陈义挺,赖钟雄,郭志雄,等.枇杷主要种类的RAPD分析[J].江 西农业大学学报,2003,25(2):258-261.
- [7]邹喻萍,葛颂,王晓东.系统与进化植物学中的分子标记[M].北 京:科学出版社,2001.10-30.
- [8] Chen DH, Ronald PC. A rapid DNA minipreparation method suitable for AFLP and other PCR application[J]. Plant Mol Bio Rep, 1999, 17: 53-57.
- [9] Khanuja SPS. Shasany AK, Darokar MP. et al. Rapid isolation of PCR amplifiable DNA from the dry and fresh sample of plant producing large amounts of secondary metabolites and essential oils by modified CTAB procedure[J]. Plant Mol Biol Rep, 1999, 17:1-7.
- [10] Porebski S,Bailey LG, Bernard R. Modification of a CTAB DNA extraction protocol for plants containing high polysaccharide and polyphenol components[J]. Plant Mol Bio Rep, 1997,15(1): 8-15.
- [11] Fang G, Hammar S, Grumet RA. Quick and inexpensive method for removing polysaccharides from plant genomic DNA[J]. Biotechniques, 1992, 13:52-54.
- [12] Wang XD, Wang ZP, Zou YP. An improved procedure for nuclear DNA isolation from silica-gel dried leaves of wild grape[J]. Plant Mol Biol Rep, 1996, 14:369-373.