Relationship between Cytoplasmic Male Sterility and Characteristics in Mitochondrial DNA in Cruciferae Plants and Garlic Plants

十字花科植物和大蒜植物细胞质雄性不育与线粒体DNA特征的关系

• 批准号: 62560032
• 负责人: OGURA Hiroshi
• 金额: $ 1.54万
• 依托单位: Kagoshima University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1987
• 资助国家: 日本
• 起止时间: 1987 至 1988
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-62560032/
• 关键词: Plant breeding; Cytoplasm; 雄性不稔性; DNA

A relationship between cytoplasmic male strility (CMS) of Cruciferae plants and variation in cytoplasmic DNAs has been investigated by a restriction endonuclrase analysis and with nine Cruciferae plants with or without CMS to elucidate the nature of CMS at the molecular level, which is useful to produce F1 hybrid plants with better characteristics in agriculture and horticulture.Chloroplasts were purified by a sucrose-gradient centrifugation method after homogenizing young, green and intact leaves of nine Cruciferae plants. Chloroplast DNA (ct-DNA) was prepared by a DNase treatment to remove a contaminated nuclear-DNA, followed by a deproteinizing treatment with proteinase K and chlorolorm-phenol mixture. Since the ct-DNA obtained thus was not digested by three types of restriction endonucleases, the nine ct-DNAs was further purified by using a commercial kit of GENECLEAN from Funakoshi Co..variation in cytoplasmic DNAs was examined by the treatment of three restriction endonucleases ( … More Hind III, EcoR I, Sal I) and consequently 1% agarose gel electrophoresis detecting total number of fragments and mobility of each fragment in the digest products.Ct-DNAs from leaves of the nine plants showed a very similar electrophoretical results each other, but a few bands of fragments were recognized to be different among them. Especially, between two species, radish and turnip, there was a distinct difference in the both electrophoretical patterns. No difference was recognized between diploid and tetraploid, and CMS species and normal species in the Cruciferae plants. In a hybrid plant, Raphanobrassica was very similar to radish at the restiction endonuclease digest patterns. Finally, it is suggested that there is no variation in ct-DNAs being related to CMA.Mitochondrial DNAs (mt-DNA) were purified from main roots of nine Cruciferae plants. Although last year they were tried to purify from seedlings, the try was unsuccessful because of difficulty to get a large numer of seeds. The yields of mt-DNAs from main roots differed much depending on the age of main roots. Some of them too low to analyze the variation in mt-DNAs. The variation in mt-DNAs is likely to be larger than it in ct-DNAs and it may be related to CMS. Less

用限制性内切酶分析了十字花科植物细胞质雄性不育性与细胞质DNA变异的关系，并对9种十字花科植物的细胞质雄性不育性进行了分子水平的研究。利用蔗糖-琼脂糖凝胶电泳法对叶绿体进行纯化，采用梯度离心法对9种十字花科植物的幼叶、绿色叶和完整叶进行分离。叶绿体DNA（ct-DNA）的制备是通过DNA酶处理以去除污染的核DNA，然后用蛋白酶K和氯仿-苯酚混合物进行脱蛋白处理。由于这样得到的ct-DNA不能被三种限制性内切酶消化，所以用Funakoshi Co.的GENECLEAN商品试剂盒进一步纯化这九种ct-DNA。通过三种限制性内切酶（ ...更多信息 Hind Ⅲ，EcoR Ⅰ，Sal Ⅰ），然后用1%琼脂糖凝胶电泳检测酶切产物中的片段总数和各片段的迁移率。尤其是萝卜和芜菁两种植物的光合作用模式存在明显差异。在十字花科植物中，二倍体与四倍体、CMS种与正常种之间均无明显差异。在杂交植株中，萝卜与萝卜的限制性内切酶酶切图谱非常相似。从9种十字花科植物的主根中提取线粒体DNA（mt-DNA），并对其进行了纯化。虽然去年他们试图从幼苗中提纯，但由于难以获得大量种子，这种尝试没有成功。主根中mt-DNA的产量因主根年龄的不同而有较大差异。其中一些太低，无法分析线粒体DNA的变异。线粒体DNA的变异可能大于ct-DNA的变异，可能与CMS有关。少