导　　师： 谭晓风

学科专业： 090702

授予学位： 硕士

作　　者： ;

机构地区： 中南林学院

摘　　要： 香榧（Torreya grandis）为我国特有的珍贵多用途经济树种，其种子为著名的干果，全身都是 宝，经济价值非常高。香榧是我国近年来发展最快、经济效益最好的经济林树种之一，优良无 性系品种越来越多。但由于良种枝条和苗木的缺乏，一些不法之徒以次充好，以假冒真的事件 在各地时有发生，因此鉴别香榧优良品种已成为当前香榧生产和良种化过程中急需解决的迫切 问题。香榧的品种分类研究仍限于形态学、解剖学的方法，有关香榧在DNA水平上的分类研 究尚未见报道。本实验试图利用RAPD技术对香榧主要栽培品种进行系统聚类分析，并进行品 种种类的划分和分子鉴别，为更加深入地研究这一珍贵树种发挥其潜在的巨大经济价值提供依 据。 本实验利用香榧种子胚乳DNA来进行RAPD分析。共有8个品种，分别为：茄榧、旋纹 榧、大圆榧、小圆榧、细榧、米榧、芝麻榧、冲杠榧。采用改良的CTAB法对香榧种子胚乳 DNA进行抽提。将抽提出来的DNA溶于TE缓冲液中，利用Du-640核酸蛋白分析仪测定DNA 的浓度。结果表明，所抽提的香榧种子胚乳DNA样品中，DNA浓度较高，但仍含有较多的蛋 白质等杂质，而且DNA的纯度不高，但由于RAPD分析对模板DNA的纯度要求不高，所抽提 的DNA样品完全能满足RAPD分析的需要。 为了提高RAPD分析的重复性，在实验过程中对RAPD分析的各个步骤都进行了反复的摸 索，特别是对RAPD的反应体系和循环条件进行了优化，最后得到了较优的适合于香榧种子胚 乳DNA进行RAPD分析的反应条件。其中RAPD反应体系为：10ng//μl DNA template 2.0μl； 10x buffer 1.5μl；2mM dNTPs 1.8μl；25mM MgCl/_2 1.44μl；10μ Torreya grandis is the special and precious economic tree with many uses in our country. Its whole body is just a gem and its seed is a well-known nut with a high economic value. In the recent years more and more improved breeds of Torreya grandis have been cultivated with the rapid productive development as one of trees with high economic benefit. But because of the lack of branches and nursery stocks of improved varieties, some cheats often happened from lawless persons by using the sham as the true. So at present the urgent problem during the course of producing and improving breeds is how to identify the ? improved breeds of Torreya grandis. Because the research on the breed classification of Torreya grandis is still limited to the research methods of morphology and anatomy, so it is very hard to find any article about the classification research at the level of DNA. In these ? experiments we try to perform the systematical analysis of assembling classes about the main cultivated breeds of Torreya grandis, and try to divide and identify the varieties and classes by using RAPD technology. The aim of this research is to provide data for the further deepen study of this precious tree species for the potential economic value. In this research the material for the analysis of RAPD is the endosperm DNA of Torreya grandis seed. there are eight breeds: qiefei, xuanwenfei, dayuanfei, xiaoyuanfei, xifei, mifei, zhimafei, chonggangfei. The DNA samples of endosperm of Torreya grandis seed which are extracted by using improved CTAB method are dissolved into TE buffer liquid for the purpose of determin

关 键 词： 香榧 标记 抽提 分子鉴别 分子分类

领　　域： [农业科学] [农业科学]