【作者】 吴臻；

【导师】 王（吉吉）之；

【作者基本信息】 陕西师范大学 ， 植物学， 2002， 硕士

【摘要】 枣原产于我国，是我国第一大干果树种和第七大果树，近年来正成为我国果树中新的发展热点。我国是世界上枣类植物资源最丰富的国家，充分发挥这一资源优势，对提高我国人民的健康水平，促进黄河流域的水土保持，大力发展国民经济具有十分重要的意义。因此，本实验试图在传统分类学基础上，借鉴前人经验，选取典型试材，利用RAPD(random amplified polymorphic DNAs)技术，尝试进行枣和酸枣种间及种下分类系统划分，探讨用RAPD进行枣品种鉴定和分类的可行性，并对RAPD结果的可靠性予以验证，建立一个严密的标准化反应体系，将这一技术应用于我省优良枣树品种的鉴定和分类上，同时进一步借助现代生物技术，为更好的开发利用名优新特枣树品种这一珍贵资源奠定基础。 实验用8个引物，得到100条扩增带，意味着对枣基因组的100个位点进行了检测。由扩增的所有条带可得到所扩增品种的指纹图谱，可以区分这50余种枣和酸枣。有3个品种产生了特有带，可作为重要分子性状用于品种鉴定和早期性状预选。在所检测品种中，除有2对间没有分开外，其他品种都可以通过两个以上的引物全部区分开。没有区分开的品种，可能是同物易名或来源于同一无性系。另外，试验中表明，只要反应体系标准化，操作规范，其稳定性是有保障的。因此，RAPD技术在枣种质鉴定中是完全可行的。实验用组间连接法构建51个枣品种及类型的遗传关系树状图，由聚类图显示，所有枣类被分为7类，第一人类包含的品种较多，有牛奶脆、蜂蜜罐、小叶枣、鸡蛋枣、清选1号、糖枣、长木枣、锥刺木枣、方木枣、狗头枣、油福水枣、白枣、木条枣、麻子枣、晋枣、耙齿牙枣、核桃纹、大荔小枣、安团，其中包含了绝大多数的陕西品种，又明显分为三组：第一组包括前4种，基本上都是鲜食品种；第二组有清选1号、糖枣、长木枣、锥刺木枣、方木枣，主要是木枣品种群；其余为第三组；第二人类有大荔水枣、酸团枣、清涧脆枣、灵宝大枣、清涧小团枣、赞皇大枣、长红枣；第三人类有绵枣、洛阳珍珠枣、河南灰枣、佳县油枣、中钻枣；第四大类有胎里红、蜂蜜罐、跌牙枣、冬枣、九月青，第五大类有天津快枣、光洋枣、酸木枣；第六人类有吴堡团枣、三变红、二秋枣、北京小枣、苹果枣、陕西龙枣；第七大类有大酸枣和交城俊枣、十月青、雪枣、无核枣。所测试的所有的枣首先聚在一起，然后再和酸枣聚在一起。从聚类图中可以看出品种间亲缘关系的远近，并鉴定出酸枣和枣的不同品种，结果说明 RAPD不仅可用于枣类品种的鉴定，而且可以用来分析品种（种）间的亲缘关系。枣100个位点的检测，对进一步研究枣的遗传结构具有重要意义。 通过实验得到以下结论： （1）在本实验室中建立了枣的 RAPD扩增体系，并找到适合枣加A提取的方法。 （2）用RAPD方法证明酸枣是大枣的野生近缘种，作为野生种，酸枣有更大的变异性，大枣由酸枣进化而来。 （3）实验用8个引物，得到lin条扩增带；意味着对枣基因组的IO0个位点进行了检测。 （4）将所测的51个枣分为7大类，枣品种间的亲缘关系与地域性有关，也与其他特征有关，如用途、果形等。更多还原

【Abstract】 Ziziphus jujuba originated from China. It’s the first dry fruit plants and the seventh fruit plants. As it ’s benefit for people’s health, Z. jujuba has been becoming a hot point in the development of Chinese fruit plants. Our country has the most abundant Z. jujuba cultivars in the world, so exploiting this resource dominance has important significance in enhancing people’s health, conservation of soil and water beside the Yellow River and developing our country’ s economy. The jujube cultivars are raised from the bud mutant. In the long history of Z. jujuba cul ture practice, many cultivars were formed. Even in Shaanxi province, there are about 130 cultivars. To understand the relationship among these cultivars has great value in the breeding of new cultivars and their generalization.Based on the traditional taxonomical research and former experience, typical cultivars was selected and used to distinguish Z. jujuba from Z. spinosus and from different Z. jujuba cultivars by RAPD (random amplified polymorphic DNA) technique. The feasibility of class if icat ion of Z. jujuba by RAPD analysis were discussed and reliability was proved. A strict standard reaction system for RAPD analysis was established, which could be used on the classification of Z. jujuba cultivars in Shaanxi province and laying foundation for breeding new high quality cultivars and spreading.In this experiment, 100 DNA bands were amplified by 8 primers, which mean 100 sites in the genome were detected. From these DNA bands, the cultivars DNA fingerprints were obtained, we could distinguish Z. spinosus and 50 kinds of Z. jujuba cultivars. 3 Z. jujuba cultivars had its own special DNA bands, which could be used as DNA molecular markers for the determination of cultivars and screening earlier characters. Except 2 pairs could not be distinguished from each other, the others cultivars could be distinguished successfully by two or more primers. The cultivars that couldnot be distinguished may be the same cultivars with different names or be derived from same strain. What’s more, the result showed that the reliability of result could be guarantied, if only the reaction system for RAPD was standard and the operation was normative. It’s mean that RAPD analysis could be used in the germplasm resources on Z. jujuba. The fifty-five jujube accessions based on their genetic distances, were clustered using unweighted pair group average cluster method (UPGMA) in a dendrogram. The genetic dendrogram was established by" between-groups linkage" method. The cluster dendrogram showed that all the cultivars could be divided into 7 class: the first class included the most of the cultivars from shaanxi province: Niunaicui, Xiaoyezao, Fengmiguan, Jidanzao,Qing-xuan, Tangzao, Changmuzao, Zhuicimuzao, Fangmuzao, Goutouzao, Youfushui- zao, Baizao, Bantiaozao, Mazizao, Jinzao, Pachiyazao, Hetaowen, Dalixiao-zao, Antuanzao, they could be divided into three groups easily; the second class included Dalishuizao, suantuanzao, Qingjiancuizao, Lingbaodazao, Qingjianxiaotuanzao, Changhongzao, Zanhuangdazao; the third class included Mianzao, Jiaxianyouzao, Luoyangzhenzhuzao, Henanhuizao, Zhongzuanzao; the fourth class included Tailihong, Dongzao, Dieyazao, Jiuyueqing, Fengmiguan; the fifth class included Tianjingkuaizao, Guangyangzao, suanmuzao, the sixth class included Sanbianhong, Erqiuzao, Beijingxiaozao, Pingguozao, Wubaotuanzao, Shaanxilongzao; the seventh class included Shiyueqing, Xuezao, Wuhezao, Jiaochengjunzao, Dasuanzao. All the Z. jujuba. Cultivars were clustered together first, then were clustered with Z. spinosus . From the dendroid cluster, We could see the relationship among them. It shows that RAPD analysis not only can be used to distinguish the Z. jujuba cultivars, but also can be used to analysis the relationship among them. The main conclusions of this paper were as follows:1. A RAPD reaction system for Ziziphus jujuba was established, and a suitable method on Z. jujuba DNA extraction was found.2. In the 50 accessions, used 8 primers and obtain更多还原