【作者】 伊艳杰；

【导师】 尚富德；

【作者基本信息】 河南大学 ， 植物学， 2004， 硕士

【摘要】 桂花是我国十大传统名花之一，原产我国，是木犀科木犀属植物，既是优良的园林绿化树种，也是著名的香花植物。桂花在长期的演化过程中，在自然选择和人工选择的双重作用下，形成了丰富的变异类型。但其系统学研究还不够深入，品种分类工作相当滞后，而且主要跟据其形态学和生理学特征进行分类。仅依此对桂花品种进行分类，难以作出客观的结论。 本文利用RAPD技术分析河南省和桂林市的桂花品种间的亲缘关系，旨在从基因组水平为桂花品种分类提供合理依据。运用层次分析法综合评价河南省的桂花品种，为今后河南省桂花品种的评选、鉴定以及进一步开发利用提供了一定的参考。主要研究结果如下： ①对影响RAPD反应的主要因素(诸如Mg²⁺浓度、dNTP浓度、DNA模板浓度以及退火温度等)，通过设浓度梯度进行了优化，获得了一个重复性高、稳定性好的RAPD反应休系。 ②利用RAPD技术对桂林市23个桂花品种的基因组多态性进行研究。20个随机引物共检测到186个位点，其中多态位点135个，占72.6％。每个引物扩增带数从5到14条不等，扩增片段大小从400到3000bp，每个品种都有其独特的标记与其他品种相区别。根据由RAPD扩增结果构建的系统树状聚类图，将这些品种划分为四个品种群。此项研究表明，以基因型而不是以表现型为基础区分桂花品种是可能的。该技术为解决桂林市的桂花品种分类问题提供了重要线索，同时也为下一步RAPD分析河南省的桂花品种奠定了一定的实验技能基础。 ③在对河南省的桂花品种进行详尽调查的基础上，选定17个桂花标准株进行定点观察，认真记录了它们的详细形态特征；并利用30个引物对其进行RAPD分析。共产生247条带，其中169条为多态性带，占总带数的68.4％，显示出桂花品种内丰富的遗传多样性。平均每个引物扩增的带数为8.07条，多态性DNA的带数为5.8条。从基于RAPD标记建立起来的树状图分析，所有供试材料的遗传距离都大于零，又都能够聚类在一起，这表明桂花品种之间既有相同遗传背景，但相互之间又存在一定差异。RAPD分析结果与传统分类学结果基本相符，首次从基因组水平为河南省的桂花品种分类提供合理依据。 ④运用层次分析法对河南省的17个桂花品种进行了生产加工应用和园林观赏方面的综合评价。分析评价中就花色、花香、花期早晚、花期长短、花径、花量、采花难易、枝条形态、树形、叶片色泽、生长势、叶稀有度等12个因子对其进行综合评价，按评分标准计算出润们有，翻价2（X）4月.月叹创七习杜忆月目卜位今食文每一品种的综合分值，根据得分将17个桂花品种品种划分为四个等级。其中演川金桂、大花金桂、朱砂丹桂、大叶黄银桂品种得分较高，与实地调查结果基本一致。总体评价表明，四季桂的品种得分相对较低，金桂品种群和丹桂品种群的大部分桂花品种在生产加工利用和园林观赏方面有粉较高的利用价值，得分也较高。 最后，在对河南省的桂花品种进行详尽地调查、R八夕D分析及综合评价的墓础上，对目前河南省在桂花品种资源利用及开发方面所存在的不足提出了几点建议，期望通过本研究能加速我省桂花品种资源的开发和利用。更多还原

【Abstract】 Osmanthus fragrans Lour, originated from China belongs to Osmanthus. As one of ten top traditional flowers in China, O. fragrans is an excellent green tree and is also famous for fragrance. During the long time of evolution, it comes out abundant variation types under influences of natural and artifical selects. The phylogenetic study is insufficient about Osmanthus fragrans Lour. The work of cultivar classification mainly accords to morphological and physiological characters, and it can’t produce subject conclusions. To offer more reasonable evidences for cultivar classification, we applied RAPD to analyze O. fragrans cultivars in Henan province and Guilin city. In addition, AHP was applied to appraise O.fragrans cultivars in our study to offer references for identifying and exploring O. fragrans cultivars in Henan province. The main results are showed as follows:(1) A repeatable and stable reaction system was obtained through optimizing influential factors. 30 better primers selected from 100 arbitrary primers were applied to amplify genomic DNA of O. fragrans Cultivars.(2)23 O. fragrans Cultivars collected from Guilin city were analyzed by RAPD markers. Among the total 186 fragments generated by 20 primers, 13S appeared to be polymorphic (72.6%). A dendrogram showing genetic relationships among them was constructed according to RAPD results. Genetic similarly coefficients ranged from 0.5171-0.9428, with the mean of 0.7619. These cultivars were divided into four cultivar groups. RAPD markers offer important clues in solving taxonornic problem of O. fragrans cultivars in Guilin city and also open up a path for analysis of O. fragrans Cultivars in Henan province.(3) 17 standard O. fragrans cultivars were fixed through thoroughly investigating mem in Henan province. Their genomic DNA were extracted and amplified by 30 primers. Total 247 bands were produced, with 169 bands were polymorphic(68.4%). 4-12 polymorphic bands could be amplified from per primer, with an average of 8.07 bands. It showed there was abundant genomic polymorphism in O. frangrans cultivars. Results from RAPD were consistent with traditionalcultivar classlification.(4) AHP was used to appraise 17 O. fragrans cultivars in Henan province about the application of landscape and productivity, color, perfum, time of blooming, diameter of the flower and other nine factors had been used in the analysis. All cultivars got marks according to the criteria. Some cultivars got higher marks, For examples, O. fragrans ’Huang Chuan Jingui’, O. fragrans ’ Dahua Jingui’ , O. fragrans ’ Zhusha Bangui’ and O. fragrans ’Dayehuang Yingui’. Results of appraisal showed most of O. fragrans cultivars in Thunbergii group and Aurantiacus group obtained higher marks. They were well used in the application of landscape and productivity.In the end, we proposed some suggestions about the exploration and application of O. fragranscuhivars in Henan province and hoped to accelerate the pace of exploring mem through our study.更多还原