辣椒细胞质雄性不育系与其保持系线粒体DNA的RAPD分析

RAPD Analyses of Mitochondrial DNAs of CMS Pepper Line and Its Maintainer Line

以辣椒细胞质雄性不育系21A及其相应的保持系21B为试材，从苗龄10～15d的黄化苗中分离纯化线粒体并提取线粒体DNA。线粒体负染后电镜观察发现，分离纯化的线粒体形态完整，呈椭球形，杂质较少。电泳检测提取的线粒体DNA的琼脂糖，结果表明，线粒体DNA纯度较高，无降解现象，浓度为20～50ng／山，可用作RAPD分析。经初步筛选，100个RAPD随机引物中有8个引物表现出多态性，经再次重复筛选，获得了辣椒细胞质雄性不育系21A线粒体DNA与雄性不育性相关的RAPD标记CMSAG3430。

The mitochondria and mtDNAs were extracted from chloresis pepper seedlings at 10 - 15 d of CMS pepper 21A and its maintainer 2lB. The mitochondria of pepper stained by PTA under the transmission electron microscrope showed intact shape, ellipsoid and less impurity. Electrophoresis detected quite pure mitochondrial DNA with no degeneration and concentration of 20 - 50 ng/μl which could be used for RAPD analysis. 100 random primers were used for analyzing the difference of mtDNA between CMS 21A and its maintainer 21B by means of RAPD technique. Among them 8 primers presented polymorphism. Furthermore, the specific RAPD marker CMSAG3430 related to CMS gene was found.