【作者】 周波；

【导师】 江海东； 张秀新；

【作者基本信息】 南京农业大学 ， 园林植物与观赏园艺， 2010， 硕士

【摘要】 牡丹是芍药属(Paeonia)牡丹组(sect.Moutan)植物,为我国特有资源,也是我国的传统名花之一。目前牡丹组植物的系统学研究主要是通过比较形态学、孢粉学、细胞学和分子生物学研究,确立了牡丹组各种的分类地位,系统调查了几个主要的栽培牡丹品种群,并对众多的品种进行了形态划分。但是有关牡丹不同来源品种间亲缘关系的比较分析研究报道较少。本文利用形态标记并结合ISSR-PCR技术,UPGMA聚类分析方法从基因组DNA扩增多态性的角度,研究部分引进牡丹品种以及国内栽培品种,不同花型、不同花色牡丹品种间的亲缘关系和遗传多样性,得到了牡丹品种之间的遗传距离图谱,为进一步研究牡丹组植物的演化关系奠定了基础。主要的结果如下：1.本研究对不同来源牡丹品种的41个形态性状进行统计分析,结果表明：牡丹组植物具有丰富的形态多样性,变异系数介于14.30%与158.99%之间,平均变异系数为44.49%。通过主成分分析,将41个性状综合为10个主成分,累积贡献率为59.97%,能反映牡丹品种形态性状的绝大部分信息。第1、3、5、6、7、8、9主成分可以合并为第1因子,主要反映花的特性(雌雄蕊瓣化情况、花瓣基部色斑情况、花径、花高、花型、花态、花色等)；第2、4、10主成分可以合并成第2因子,主要反映叶的特性(新枝长、复叶类型、叶长、叶宽、侧小叶形状、小叶叶尖形状等等),主成分分析结果支持聚类分析结果。基于形态性状的聚类分析把这些品种聚为两类：第一类绝大多数为国外品种,其特点为叶表面无色晕、花径较大、花瓣无色斑、花期晚、花朵向上；第二类同时包含了国内和国外品种,但它们可以通过进一步的聚类加以区分,其特点为叶表面无色晕、花径较小。聚类结果显示通过形态多样性分析可以把不同来源的牡丹品种分类。在牡丹育种中,选择来源不同的亲本,有可能得到性状优良的杂交后代。2.采用ISSR-PCR分子标记技术对供试牡丹品种的遗传多样性进行了分析,从55条引物中筛选出能获得清晰条带且多态性好的19条分别对供试材料基因组DNA进行扩增,共获得188条清晰可辨的条带,其中多态性带177条,多态位点百分率为94.1%,多态性较高。POPGENE软件计算结果表明,供试牡丹品种平均有效等位基因数为1.514,平均Nei’s基因多样性指数为0.3085,平均Shannon信息指数为0.4713；应用NTSYS软件计算得到各品种间的相似系数介于0.3294和0.7744之间,平均为0.5722。UPGMA法将供试品种分成2个类群,聚类结果与来源及花型花色相关。两种标记方法对不同来源牡丹品种遗传多样性的评价相近。更多还原

【Abstract】 The germplasm of Paeonia sect.Moutan was endemic to China. China is not only the distribution center of wild species of Sect.Moutan, but also has cultivated numerous cultivars of P. suffruticosa. The phylogenetic research work of Sect.Moutan was mainly concentrated in the taxonomy of species of Sect.Moutan and morphologic classification of cultivars by morphologic observation, sporo-pollen analysis, cytologic and molecular biologic technique. But few focused on the phylogenetic relationship and diversity about species of different origin expecialy with molecular phylogenetic techinology. This paper reported the relative research work by morphology and ISSR analysis. This experiment enrichied the molecule phylogenesis studies of Paeonia Suffriticosa, the dendrogram of descendibliy was obatined, which gave the foundation for the further research, using this relationship can provide predicitions for new cultivars breeding of tree peony and contribute it to the tree peony industry in China. The major results were as follows:1In this paper, forty-one morphological characters were investigated in93cultivars of Peony. The results showed that the morphological diversity was high among cultivars. Coefficient of variation based morphological traits ranged from14.30%to158.99%with the average of44.49%. The principal component analysis showed that the41characters were integrated into10principal components.10principal components represented59.97%of variance, containing majority information of all index. Further more, the first, third, fifth, sixth, seventh, eighth and ninth components can be combined into the first factor, representing for flower characters (stamen state, petaled anther color, petaled anther form, petal basal spot form, petal basal spot color, flower form, pedicel state, petaled pistil color, filament color, flower color, stigma color, flower across length, flower height, plant height). And the second, fourth and tenth components can be combined into the second factor, representing for leaf characters (leaf tip shape, length of fresh-branch, top-leaf top, compound leaf type, length of leaves, width of leaves, side-leaf form). The principal component analysis result supported cluster analysis result. Based on the morphological data, these cultivars of Peony were clustered into2groups. The first group was characterized by leaf faint, big flower across length, no petal basal spot, late florescence and pedicel upwards, most cultivars of which are foreign cultivars. The second group was characterized by no leaf faint and small flower across length, including foreign and Chinese cultivars, which can be classified by further clustering. Cluster analysis result showed that peony cultivars with different origin can be classified by morphological diversity analysis. Optional progeny were achieved by crossing from different country parents in breeding of Peony.2ISSR-PCR was used to detect the genetic diversity and relationship of these peony cultivars. ISSR fingerprinting amplified by19ISSR primers revealed a total number of188unambiguous bands, of which177ones were polymorphic and the polymorphism frequency was94.1%. As analyzed by POPGENE1.31, the average value of effective number of alleles, Nei’s gene diversity and Shannon’s information index were1.514,0.3085and0.4713, respectively. As analyzed by NTSYS, the similarity coefficient between cultivars ranged from0.3294to0.7744, the average value is0.5722. These peony cultivars were divided into2groups by UPGMA (unweighted pair group method with arithmetic average) based on similarity coefficient. The genetic relationship of these cultivars was related to their origin, flower form and flower colour. The results of the evaluations of genetic diversity in different original peony cultivars based on the two methods were similar.更多还原