【作者】 黄丽峰；

【导师】 黄儒珠；

【作者基本信息】 福建师范大学 ， 生物化学与分子生物学， 2007， 硕士

【摘要】 应用RAPD和ISSR标记对木兰科6属20种植物的遗传多态性和亲缘关系进行分析，比较属间、属内的遗传差异，并结合前人的研究结果，对木兰科6属及属下类群的系统位置进行探讨，旨在为木兰科植物的系统分类及其它研究提供实验依据。研究结果如下：（1）建立了适合于木兰科6属20种植物DNA提取的改良CTAB法Ⅰ，用该法提取的DNA能满足RAPD和ISSR分析的要求；（2）采用单因素结合均匀试验设计优化得到适合于木兰科6属20种植物RAPD和ISSR分析的PCR体系。优化的RAPD-PCR体系为：Mg²⁺3.0 mmol／L、dNTP 0.15 mmol／L、引物0.40μmol／L、模板DNA20 ng／20μL、Taq DNA聚合酶1 U；优化的ISSR-PCR体系为：Mg²⁺ 1.00mmol／L、dNTP 0.20 mmol／L、引物0.40μmol／L、模板DNA 40 ng／20μL、Taq DNA聚合酶1 U；（3）分别从112条RAPD引物和99条ISSR引物中筛选出14条RAPD引物和12条ISSR引物用于RAPD和ISSR分析。14条RAPD引物共检测到287个位点，其中多态位点占99.30％，Nei’s基因多样度为0.3591，Shannon信息指数为0.5334；12条ISSR引物共检测到222个位点，且222个位点都是多态的，Nei’s基因多样度为0.3477，Shannon信息指数为0.5218。RAPD和ISSR分析都表明木兰科6属20种植物具有丰富的遗传多态性；（4）基于RAPD和ISSR扩增条带数据建立了相应的Nei’s（1979）遗传距离矩阵，以及UPGMA树图。基于RAPD标记和ISSR标记构建的树图，两者趋势大致相近，但不完全相同；（5）综合木兰科6属20种植物的RAPD和ISSR分析结果，支持鹅掌楸属、木莲属、拟单性木兰属和观光木属的独立；建议重新界定木兰属的范围，并成立玉兰属；建议取消含笑属下亚属和组的划分，设为含笑属。更多还原

【Abstract】 Randomly amplified polymorphic DNA （RAPD） and inter-simple sequence repeat （ISSR） markers were used to analyse the genetic diversity and relationships of 20 species in 6 genera from Magnoliaceae and compare the genetic difference among generas with inner genera, combining the other research results, the systematic of 20 species in 6 genera was primarily discussed. The purpose of this study was to provide scientific basic data for systematic and other research on Magnoliaceae.The results showed as follow: （1） The method of improved CTABⅠequal to extract Magnoliaceae genomic DNA was developed, because the extracted DNA by the method was suitable for RAPD and ISSR analysis. （2） Single factor design combing uniform design was used to obtain optimum scheme on reaction system for RAPD and ISSR of Magnoliaceae. The obtained optimum scheme on reaction system for RAPD: Mg²⁺ 3.00 mmol/L、dNTP 0.15 mmol/L、primer 0.40μmol/L、template DNA 20 ng/20μL、1U Taq DNA polymerase; The obtained optimum scheme on reaction system for ISSR: Mg²⁺ 1.00 mmol/L、dNTP 0.20 mmol/L、primer 0.40/μmol/L、template DNA 40 ng/20μL、1 U Taq DNA polymerase。（3） The analysis of RAPD marker showed that: fourteen RAPD primers amplified a total of 287 scorable markers, percentage of polymorphic loci is 99.30%, Nei’s gene diversity and Shannon’s information index were 0.3591 and 0.5334, respectively. The analysis of ISSR marker showed that: twelve ISSR primers amplified a total of 222 scorable markers, all of which was polymorphic loci, Nei’s gene and Shannon’s were 0.3477 and 0.5218, respectively. The results of RAPD and ISSR showed that the genetic diversity of 20 species in 6 genera from Magnoliaceae was enriched. （4） The matrixes of genetic distance were established on RAPD and ISSR fingerprint, respectively. The results of UPGMA duster analysis based on RAPD and ISSR fingerprint had the approximate trend, but not the same. （5） The synthetical comparison of RAPD and ISSR sustained the independency of Liriodendron, Manglietia, Parakmeria and Tsoongiodendron, and advised to redefine the range of Magnolia, subgen. Yulania indepent from Magnolia, then established Yulania, and abolished the division of subgenera and section Michelia, and set up Michelia.更多还原