【作者】 戴思慧；

【导师】 钟晓红；

【作者基本信息】 湖南农业大学 ， 果树学， 2004， 硕士

【摘要】 李及其近缘种栽培历史悠久，分布广泛，是一类重要的经济果树。由于品种间的引入导致许多同名异物或同物异名的现象比较普遍，影响李种质资源的评价和利用。以往多从形态学、孢粉学、同工酶等方面研究李及其近源种种质的分类演化，取得了一定的成果。但还存在许多分歧，在技术手段上也存在着一定的局限性。本研究通过RAPD技术，从DNA水平上研究李及其近缘种的分类、遗传演化关系。主要结果如下： 1．用高盐低pH法、改良SDS法和CTAB法三种方法提取李的幼嫩叶片基因组DNA，结果表明高盐低pH法是较好的提取李基因组DNA的方法。增加盐的浓度能够避免提取的基因组DNA呈凝胶状，有利于多糖的去除；在基因组DNA提取过程中，干粉PVP和巯基乙醇一同使用去除酚类物质的效果较好。 2．从200个随机引物中筛选出具有多态性强、带型清晰的引物18个，在分析李及其近缘种种质时均得到成功应用。 3．用18个随机引物扩增的指纹图谱分析所得到的聚类树状图表明郁李与李的遗传距离为0.0491，表现出很近的亲缘关系；而与樱桃的遗传距离为0.3761，表现出较远的亲缘关系，因此，应将郁李视为李的一个种。 4．在遗传距离0.1978～0.2999的水平上，供试的41份材料聚分为4类：桃类、杏梅类、樱桃类和李类。从指纹图谱来看，各类有其特有的特征谱带。因此，本研究结果支持将核果类果树分成不同的属，即桃属、杏梅属、樱属和李属。 5．在遗传距离为0.1201～0.1326时，所有材料分为11类，桃为第1类：杏聚分为第2、3类；花梅为第4类；美人梅为第5类；樱桃为第6类；李聚分为第7-11类。其中奈李为第7类；红叶李第8类；澳得罗达、玫瑰皇后、美丽李、黑宝石聚为第9类；艳红和烟花为第10类，其余的李聚为第11类。 6．从分子手段证实奈李为李的一个变种，而非李与桃的杂种。更多还原

【Abstract】 Prunus have a long-term of being cultivated all round the world, and are important fruit tree that they can get big wealth. A common problem confronting prunus taxonomists and horticulturist in selecting and utilizing is the homonym and synonym caused by prunus germplasm resources exchanging, which is not benefit for identification and utilizing of prunus germplasm resources. Lots of works have been done in morphology, pollen and isoenzyme with some performance about the identification, taxonomy and evolution of prunus. But there are still some divergence in result and limitation in technology. RAPP studied genetic relationship in germplasm resources of prunus. The main results are as follows:1.Based on the analysis of three isolation methods for genomic DAN of prunus, the results showed that high-salt, low-pH method is optimal for isolation method. Tissue extraction with SDS in the presence of a high salt concentration suppressed co-precipitation of polysaccharides and DNA. So DNA preparations were free of gelatinous substances. During the total DNA isolation, using together PVP powder and 2-mercaptoethanol is benefit for eliminating phenols.2.Eighteen decameter arbitrary primers, which had polymorphisms and dear bands, were selected out of two hundred;, the primers were successfully amplified in analysis of prunus germplasm.3.Cluster tree of materials in Prunus by eighteen decamer arbitrary primers amplifying bands showed that Prunus japonica Thumb had higher genetic similarity in prunus salicina and was far relationship from P. lannesiana .So Prunus japonica Thumb should be belonged to prunus salicina .4.At the genetic distance level of 0.1978~0.2999, all of materials are classified 4 kinds. Besides every kind has a specific band. So the results support that stone fruit trees are classified for P. persica, P. armeniaca and P. mume, P.cerasus and Prunus salicina.5.When genetic distance is 0.1201-0.1326, all of materials are classified 11kinds. Peach is the first; apricot have 2 kinds; huamei is the forth; meirenmei is the fifth; cherry is the sixth; plum is classified 5 kinds. Naili and hongyeli is different kind. Aodeluodali, meiguihuanghouli , meilili and heibaoshi is the same kind. Yanhong and yanhua cluster a kind. The rest cluster the same kind.6.NaiLi is a Variety of primus salician, not a hybrid of P.salician and P. Persian by molecular biological methods.更多还原