【作者】 李彦青；

【导师】 何龙飞； 李志刚；

【作者基本信息】 广西大学 ， 作物栽培学与耕作学， 2004， 硕士

【摘要】 本研究应用RAPD标记，选用10个随机引物对36份杂交水稻亲本材料进行了多态性分析，共检测到106条带。81条多态性带。聚类分析结果表明，所有供试材料可以被明确的区分。 为了得到一个稳定的RAPD反应体系，首先对影响反应体系稳定性的因子进行了优化，优化结果为：DNA模板用量10～60ng；Mg²⁺浓度2.0mmol／L；dNTPs浓度1.5～2.0mmol／L；引物浓度1.2～2.0μmol／L；Taq聚合酶用量0.8～1.2U。然后对40条引物进行筛选，结果得到10条具有重复性好、条带清晰、多态性高的引物，即S227、S228、S229、S290、S358、S400、S490、S1405、S1422、S1492。 用NTSYS-pc（version 2.1）软件包中的Nei&Li法的计算方法计算36个亲本材料的遗传相似系数，用UPGMA法聚类分析，用NJTREE程序构建亲本间的树状聚类图。36个亲本材料的遗传相似系数和树状聚类图结果表明，所有的供试材料，它们之间的相似系数较大，遗传差异较小。三系杂交水稻亲本间仍然有较为明显的多态性差异，即使是遗传背景相近的不育系与保持系间也存在多态性。更多还原

【Abstract】 Twelve rice male sterile lines, 9 maintainerblines and 15 restorer lines were analyzed by RAPD with 10primers.altogether,106 fragments were generated,of which 81 detected polymoprphism among rice lines tested.Cluster analysis showed that all lines could be uniqeuely distinguished by RAPD marker.In order to obtain a stable RAPD reaction system, we optimized the reaction conditions based on experimental results. The results showed that the reaction system , which included 10~60ng DNA, 2.0mmoI/L Mg2+, 1.5~2.0mmol/L dNTPs, 1.2~2.01 mol/L random primers and 0.8~1.2U Taq polymerase, was good for our experiments. Ten primers, which produced repeatable, clear and rich amplified polymorphism in the RAPD analysis, were obtained by screening 40commercial primers. They are S227,S228,S229,S290,S358,S400,S490,S1405,S1422and S1492.Based on the pair-wise comparsons of amplification products the genetic distance was calculated using Nei&Li’s genetic similarity coefficient and a dendrogram was constructed using an un-weighted pair group method with arithmetical averages (UPGMA),the result that the genetic similarity coefficient was very identical and the genetic difference of 36 rice lines was very small, their genetic polymoprphism was distinguishable,even the genetic backdrop among the male sterile lines and maintaier lines was very similarity,their genetic polymoprphism was also distinguishable.更多还原