摘要
目的：获得稳定性、重复性好的RAPD-PCR反应体系,并构建血满草DNA指纹图谱和进行遗传多样性分析。方法：以血满草3个居群11个个体的幼叶为材料,CTAB法提取基因组DNA,从模板浓度、引物浓度、d NTPs浓度和Taq DNA聚合酶的用量,构建最佳的RAPD-PCR反应体系,通过扩增带型的差异构建其DNA指纹图谱,利用Popgen32、NTSYS2进行遗传多样性分析。结果：3个RAPD引物扩增血满草3个居群11个样品共获得27条可靠、清晰和重复性高的条带,其中17条是多态条带。引物SBSA5和SBSA11扩增条带组合可以构建11个样品的个体特异DNA指纹图谱。供试血满草居群间遗传距离在0.1493~0.2312之间;居群内的遗传多样性分别为0.1102、0.0153、0.2294。遗传距离和相似性聚类分析表明,样品间的遗传距离与居群的地理分布关系不明显。结论：RAPD分子标记在构建血满草DNA指纹图谱是可行的,由其构建的指纹图谱可以将供试个体相互区分鉴别出来。无论是居群间还是居群内,供试血满草的遗传多样性均较低,居群间的遗传分化较小,可能还在属于同一个大的居群。
Objective： To obtain stability and reproducibility RAPD- PCR reaction system,and build DNA fingerprinting and analyze genetic diversity of Sambucus adnata L. Basing on RAPD markers. Method： Genomic DNA was extracted by CTAB from three populations,11 individual leaves,and the optimal RAPD- PCR was constated from four factors,i. e. DNA template,primer,d NTPs and Taq DNA polymerase concentration. Using optimal reaction system,the DNA fingerprinting was constructed by different types and genetic diversity was analyzed by Popgene32 and NTSYS2. Result： Three RAPD primers with high polymorphism and repeatability were successfully screened out from 20 candidate primers. 27 reliable,clear and reproducible bands with 17 polymorphic ones were amplified from three populations,11 samples using three RAPD primers. And individual- special DNA fringerprinting was constructed by amplified bands using primer SBSA05 and SBSA11. The genetic diversity within S. adnata samples was low and genetic distance among populations was 0. 1493- 0. 2312,and genetic diversity within populations were 0. 1102,0. 0153,and 0. 2294,respectively. Genetic distance and similarity cluster analysis showed that the relationship between genetic distance and geographic distribution of samples was not obvious. Conclusion： RAPD molecular markers to build DNA fingerprinting in S. adnata is feasible,and building its fingerprint can be distinguished from each individual identified. Whether among or within populations,genetic diversity of S. adnata samples for the test were low,and smaller genetic differentiation was among samples,it indicated that all samples for the test may belong to the same larger populations.
出处
《生物技术》
CAS
CSCD
北大核心
2014年第5期48-51,共4页
Biotechnology
基金
国家自然科学基金项目("壳斗科栎属川滇高山栎分子谱系地理学研究"
No.31060031)
云南民族大学民族药资源化学国家民委-教育部共建重点实验室开放基金项目("罗平布依族药用植物资源的民族植物学调查"
No.MZY1301)资助
关键词
接骨木属
民族药
遗传多样性
聚类分析
Sambucus L. Ethnic medicine Genetic diversity Cluster analysis
