摘要
利用CDDP(Conserved DNA-derived Polymorphism)分子标记方法,对47个不同花色紫斑牡丹品种的基因组DNA进行遗传多样性分析。结果显示:(1)采用正交实验设计,对影响紫斑牡丹CDDP-PCR反应的引物浓度、DNA模板浓度两因素进行优化,优化后紫斑牡丹CDDP-PCR反应体系为:总反应体系20μL,其中包括2×Es Taq MasterMix酶(含染料)10μL,10pmol/μL引物1.5μL,15ng/μL DNA模板4μL,ddH2O 4.5μL。(2)21条引物中,有19条产生了清晰的条带。19条引物共扩增出112条带,其中多态性条带97条,占总条带数的86.61%,多态性比例高。(3)UPGMA聚类显示,47个紫斑牡丹品种的聚类大体按照花色聚类,品种间遗传相似系数值在0.72~0.95,说明品种间相似度高,差异程度小。研究表明,CDDP分子标记方法适用于紫斑牡丹遗传关系分析,且特异条带的产生为深入研究紫斑牡丹品种鉴别提供理论依据。
CDDP molecular marker was used to analysis genetic diversity of 47 Paeonia rockii DNA.The results showed that:(1)the orthogonal experimental design was used to optimize CDDP amplification system on P.rockii resources in primers and template DNA.A suitable CDDP reaction system was established:the CDDP reaction mixture,with a total volume of 20μL,consisted of 10μL of 2×Es Taq MasterMix,1.5μL of 1.0 pmol/μL primer,4μL of 15 ng/μL DNA and 4.5μL of ddH 2O.(2)19 of 21 primers yielded 112 bands,97 were polymorphic which accounted for 86.61%of all bands.(3)The results of UPGMA showed that:47 P.rockii were grouped together with the same color in general.The similarity index value was 0.72~0.95 which showed the similarity degree among 47 P.rockii was high and the difference degree was low.In total,CDDP molecular marker was suitable for analyzing genetic diversity of P.rockii.The emergence of specific bands could provide theoretical basis for further study the identification of P.rockii.
作者
陈燕
李婉茹
唐红
CHEN Yan;LI Wanru;TANG Hong(Gansu Agriculture University Academy of Forestry,Lanzhou,730000)
出处
《西北植物学报》
CAS
CSCD
北大核心
2018年第10期1823-1831,共9页
Acta Botanica Boreali-Occidentalia Sinica
基金
甘肃省农牧厅生物技术专项(GNSW-2014-7)
关键词
紫斑牡丹
CDDP
分子标记
引物筛选
遗传多样性
Paeonia rockii
CDDP(conserved DNA-derived polymorphism)
molecular marker
primer screening
genetic diversity
