【作者】 夏惠；

【导师】 张振文；

【作者基本信息】 西北农林科技大学 ， 果树学， 2005， 硕士

【摘要】 欧洲葡萄是葡萄属植物中栽培价值最高的种,拥有许多优良品种（5000 个以上）,广泛的分布于世界各地。世界上著名的鲜食、酿造及其它加工品种大多是本种或本种的杂种。由于在长期的进化过程中和栽培历史中形成了许多种群和品种群,且葡萄种内变异很大,加上种间的自然杂交及世界范围内对葡萄的广泛繁殖,使得葡萄的分类鉴定十分困难。本试验利用RAPD技术对68 个葡萄品种（类型）和21 个葡萄营养系（品种）的遗传多样性进行分析,并得到以下结论: 1. 比较了两种DNA 提取方法,SDS 法与CTAB 法提取的DNA 都可用作RAPD模板,但CTAB 法操作相对简单,用时少,提取的DNA纯度也较高。2. RAPD 技术在本试验条件下的最优化体系为:1×反应缓冲液、0.2mM dNTP、2.0mM Mg²⁺、1.5U Taq DNA聚合酶、0.2μM 引物、20～40ng 模板DNA;反应条件为:94℃预变性7min,94 ℃1min,36 ℃1min,72 ℃2min,45 个循环;72℃10min。3. 对56 个引物进行筛选,得到18 个多态性好、重复性好的引物,用来对供试材料进行分析。4. 用RAPD方法对68 个葡萄品种进行聚类分析,结果可将供试品种分为九类:刺葡萄、山葡萄和毛葡萄各为一类;乍娜、京秀、玫瑰香、早玫瑰、葡萄园皇后、山东早红、莎巴珍珠、早金香为第四类;第五类有白诗南、白玉霓、缩味浓、琼瑶浆、佳丽酿、雷司令等18 个品种,这些葡萄品种大都起源于法国、西班牙、德国、奥地利等国家,在分类学上属于西欧品种群;第六类:泽香、白鸡心、魁宝、绯红、力扎马特、马奶子、红脸无核等16 个品种,在分类学上属于东方品种群。第七类包括,红什佳美、粉红玫瑰、白什佳美、小白玫瑰,它们起源于希腊、保加利亚一带,在分类学上属于黑海品种群。第八类:天秀、黑奥林、京亚、露都蓓蕾、黑潮、紫珍香、康拜尔早生、苿莉、红蜜、井川1014、红瑞宝等13 个品种,它们是巨峰系的欧美杂交种。第九类:巨峰、白富士、京超、佐藤,也是巨峰系的欧美杂交种。5. 分别用RAPD法和形态法对21 个葡萄营养系（品种）进行聚类分析,结果表明,只用形态法无法将营养系正确区分开,而使用RAPD分子标记的方法可将主栽培品种的营养系区分开。聚类结果表明,品丽珠、梅尔诺与赤霞珠关系较近。更多还原

【Abstract】 Vitis vinifera L. is the predominant commercial specie in all of Vitaceae Lindl plants, including many high-grade varieties （over 5000）. Most of high quality table grapes、wine grapes and other processing varieties are belong to this specie or its hybrids. During long history of evolution and grapevine growth, many specie groups and variety groups has formed. Because aberrance in specie occurres all along and grapevines are propagated by cutting, so these often make grape varieties identification difficult. Genetic diversity of 68 grape varieties and 21grape clones has been researched and the results are as flows: I. Compared two methods of DNA extraction:SDS and CTAB. DNA sample extracted by these two methods were all of good quality for RAPD. Comparatively, the method of CTAB was simpler、more efficient and DNA sample obtained has higher purity. II. Optimized RAPD reaction system and suitable conditions was obtained: 1×buffer, 0.2mM dNTP, 2.0mM Mg²⁺, 1.5U Taq DNA polymerase, 0.2μM primer, 20～40ng template DNA;reaction condition:94 ℃7min; 94℃1min,36℃1min,72 ℃2min,45 cycles; 72℃,10min。III. 56 primers were used for amplification, among which 18 primers were selected for RAPD analysis. These primers could amplify stable and polymorphic RAPD bands. IV. Tree diagram was constructed based on Euclidean distance matrix using UPGMA （unweighted pair group method using arithmetic average） cluster analysis. It was showed on the dendrogram that 68 varieties belong to 9 groups: V. davidii Foex is the first group; V. amurensis Rupr. is the second group; V. quinquangularis Rehb is the third group; Zana, Jingxiu, Muscat Hamburg, Zao jinxiang, Zao Meigui, Queen of Vineyard, Shandong Zhaohong and so on （8 varieties） formed fourth group;Sauvignon Blanc, Gewurztraminer, Carignan, Riesling, Chenin Blanc, Ugni Blanc and so on （18 varieties） formed fifth group, these blong to Proles occidentalis Negr; Zexiang, Baijixin, Kuibao, Feihong, Rizamat, Manaizi, Blush Seedless and so on （16 varieties） formed sixth group, these blong to Proles orientalis Negr; Red Gamay, Muscat Rose, Camay Blanc, Muscat Blanc （4 varieties） formed seventh group, these blong to Proles pontica Negr ; Tensyu, Blank Olympia, Beniizu, Shina bailey, Kuroshio, Zi Zhenxiang, Campbell Early, Maciji, Honey Red, Ikawa 1014, Ikawa 1025, Benizuiho, Jingya and so on （13 varieties） formed eighth group; Kyoho, Sato, Jingchao, White fuji （4 varieties） formed ninth group. V. Using morphological method and RAPD method respectively to clustering analysis 21 grape clones, the results showed that only using morphological method can’t distinguish these clones, but using RAPD method can distinguish them. clustering analysis indicated that Cabernet Franc, Cabernet Sauvignon and Molet have close relationship.更多还原