摘要
野生玫瑰中多糖多酚含量较高,DNA提取较为困难,本试验在植物基因组DNA CTAB提取法的基础上,通过延长水浴时间、调整氯仿-异戊醇（24∶1）抽提次数为2~3次等一系列方法提高野生玫瑰基因组DNA的提取纯度。以2×Es Taq Master Mix（含染料）10μL,40 ng/μL DNA模板1μL,10 pmol/μL引物1μL,dd H2O 8μL为CDDP-PCR反应体系,对21条CDDP引物进行多态性筛选。结果表明,用改良的CTAB法提取的野生玫瑰基因组DNA纯度较高;21条CDDP引物中有16条对野生玫瑰品种的鉴别能力较高。本研究为野生玫瑰CDDP分子标记及后续各种分子生物学研究奠定基础。
DNA extraction of Rosa rugosa is difficult for their high contents of polyphenol and polysaccharide. Here a modified CTAB method for Rosa rugosa DNA extraction was presented through extending water- bath time,appropriately adjusting chloroform isoamyl alcohol（ 24∶1） extraction time to 2 ~ 3 times and etc. Twenty- one CDDP polymorphic primers were screened with reaction system containing 10 μL of 2 × Es Taq Master Mix（ including dye）,1 μL of 40 ng/μL DNA template,1 μL of 10 pmol/μL primer and 8 μL of dd H2 O. The results showed that the genomic DNA of Rosa rugosa extracted with modified CTAB method owned high purity,and 16 CDDP primers had higher discriminating ability on Rosa rugosa. This research provided bases for CDDP molecular markers and subsequent molecular biology researches of Rosa rugosa.
出处
《山东农业科学》
2016年第11期13-17,共5页
Shandong Agricultural Sciences
基金
山东省农业良种工程项目"林木种质资源收集保护与评价"[鲁农良字(2010)6号]
