【作者】 尹燕雷；

【导师】 赵兰勇； 苑兆和；

【作者基本信息】 山东农业大学 ， 农业推广， 2008， 硕士

【摘要】 石榴(Punica granatum L.)属石榴科(Punicaceae)石榴属(Punica L.)植物,又名安石榴、若榴、天浆等。原产中亚,是一种古老的温带落叶果树。全世界栽培的石榴都属于石榴属中的石榴(Punica granatum L)这个种,但由于不同栽培石榴品种对生态环境适应能力的差异性与选择性,加上驯化栽培和人为选择,形成了不同地理生态条件下的栽培群体。本文利用分子系统学的原理和方法,采用荧光AFLP标记技术,分析了山东主要栽培石榴品种之间的遗传多样性,为山东优良石榴品种选育提供分子依据。主要研究结果如下:1、山东25个石榴品种,即‘大马牙甜’、‘青皮软籽’、‘三白甜’、‘大红皮酸’、‘重瓣玛瑙’、‘牡丹石榴’、‘白皮酸’、‘冰糖冻’和‘重瓣白石榴’、‘小红皮甜’、‘青皮谢花甜’、‘青皮岗榴’、‘墨石榴’、‘月季石榴’、‘红皮马牙甜’、‘枣庄短枝红’、‘巨籽蜜’、‘鲁峪酸’、‘小青皮甜’、‘大青皮甜’、‘大红皮甜’、‘大青皮酸’、‘泰山三白石榴’、‘泰山红’和‘大红袍’,的形态特征进行初步的统计。结果表明,山东石榴品种的果皮颜色和花瓣颜色差异较大,并且无显著相关性;果实风味以甜味为主(占78%),花瓣以单瓣花为主,同时有部分复瓣花;2、利用荧光AFLP标记技术,从64对EcoRI/MseI引物中选出8对引物,25个石榴品种共扩增出1728条带,其中多态性位点数为722条,占总数的41.78%。不同的引物组合扩增的多态性位点数目和多态位点百分比明显不同,8对引物组合扩增的多态性位点数从70(E-AAG/M- CAT)到126(E-AAG/M- CTA)条不等,平均多态性位点数为90.25条;E-AAG/M-CTA引物组合的多态性百分比最高P=58.33%,E-AAG/M- CAT引物组合的多态性百分比最低P=32.41%,平均多态性百分比为41.78%。说明供试品种在DNA水平上酶切位点的分布存在广泛的变异。3.本文利用POPGENE version 1.32软件对各位点的观测的等位基因数目(Na)、有效等位基因数目(Ne)、Nei基因多样性(H)和香农信息指数(I)进行了统计分析。观测的最高等位基因数目为1.5833(E-AAG/ M-CTA),最低为1.3241(E-AAG/ M-CAT),所有位点平均的等位基因数目为1.4178;有效等位基因数目最高为1.2618(E-AAG/ M-CTA),最低为1.1610(E-AAG/ M-CAA),平均有效等位基因数目为1.1874;各位点的Nei基因多样性为0.0727(E-AAG/ M-CAT)-0.1603(E-AAG/ M-CTA),平均为0.1133;香农信息指数为0.1166(E-AAG/ M-CAT)-0.2486(E-AAG/ M-CTA),平均为0.1752,方差分析显示H和I在多数位点具有显著性差异,少数位点具有极显著差异。说明山东石榴遗传多样性较为丰富。4.基于Lei& Li相似系数利用UPGMA法进行聚类分析,在相似系数0.786处将25个石榴品种划分为四大类,进一步用AFLP技术证明山东石榴品种遗传多样性丰富。本研究根据AFLP结果探讨了山东石榴的亲缘关系,并对进一步的品种选育进行了展望。更多还原

【Abstract】 Punica granatum L., alternate name‘An Shiliu’,‘Ruo Liu’,‘Tian Jiang’which belongs to Punicaceae, Punica L., occurred originally Central Asia. It is a deciduous fruit tree in temperate zone. Punica granatum L.of all over the world belongs to this species. Owing to the differences and selectivities to the adaptability of different ecological environments and domesticating cultivation and artificial selection, cultivation groups of different geological environments were coming into being. The genetic diversities of pomegranate cultivars in Shandong were analyzed using molecular systematical theories and methods by AFLP. This paper provided the basis for the selection and breeding of Shandong pomegranate cultivars.The main results were as follows:1. The morphological characteristics of pomegranate was analyzed from 25 cultivars including‘Damayatian’,‘Qingpiruanzi’,‘Sanbaitian’,‘Dahongpisuan’,‘Chongbanmanao’,‘Mudanshiliu’,‘Baipisuan’,‘Bingtandong’,‘Chongbanbaishiliu’,‘Xiaohongpitian’,‘Qingpixiehuatian’,‘Qingpigangliu’,‘Moshiliu’,‘Yuejishiliu’,‘Hongpimayatian’,‘Zaozhuangduanzhihong’,‘Juzimi’,‘Luyusuan’,‘Xiaoqingpitian’,‘Daqingpitian’,‘Dahongpitian’,‘Daqingpisuan’,‘Taishansanbaishiliu’,‘Taishanhong’,‘Dahongpao’. The results showed that the differences between peel colour and petal colour were great and there were no significant correlations. The main flavour of fruits was sweetness. The simple flower hold the most portion of the flowers and then double-petalled blossoms.2. The AFLP maker technique was carried out with eight pairs primers from 64 pairs. The total 1728 bands were amplified in 25 pomegranate and the polymorphic loci were 722 which had a proportion of 41.78%. The number and the percentage of polymorphic loci amplified by differert primers combination were obviously differernt. The polymorphic loci amplified by 8 pairs primers ranged from 70(E-AAG/M- CAT)to 126(E-AAG/M- CTA), and the main polymorphic loci were 90.25. The percentage of polymorphic loci amplified by E-AAG/M- CTA combination were the highest (P=58.33%) while E-AAG/M- CAT the lowest (P=32.41%). The main percentage of polymorphic loci was 41.789%. The results indicated that distributions of the restriction sites at the DNA level of these cultivars had wide variations.3. The Na, Ne, H and I of observed locis were statistical analyzed using POPGENE version 1.32 software. The highest number of observed alleles were 1.5833(E-AAG/ M-CTA)and the lowest were 1.3241(E-AAG/ M-CAT). The average number of alleles were 1.4178. The highest effective number of alleles were 1.2618(E-AAG/ M-CTA)and the lowest were 1.1610(E-AAG/ M-CAA). The average effective number of alleles were 1.1874. The gene diversities of all allels were 0.0727(E-AAG/ M-CAT)-0.1603(E-AAG/ M-CTA)while the main was 0.1133. The I was 0.1166(E-AAG/ M-CAT)-0.2486(E-AAG/ M-CTA)while the main was 0.1752. Variance analysis showed that H and I had differences at the most locis while significant differences at few locis. The results showed that pomegranate of Shandong had rich genetic diversities.4. The cluster analysis of the Lei & Li was carried out by UPGMA. 25 cultivars were divided into four groups at 0.786 level of similarity coefficient. The genetic diversities of Shandong pomegranate were further proved by AFLP.The genetic relationships of pomegranate in Shandong were discussed according to the AFLP results. The prospects of selection and breeding of Punica granatum L. cultivars were also put forward in this paper.更多还原