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

通过限制性内切酶分析，研究了十字花科植物细胞质雄性不育（CMS）与细胞质dna变异之间的关系，从分子水平上阐明了CMS的性质，为培育具有较好农艺性状的F1杂交植物提供依据。对9种十字花科植物幼叶、绿叶和完整叶片进行均质处理，采用蔗糖梯度离心法纯化叶绿体。采用脱氧核糖核酸酶（DNase）去除受污染的细胞核DNA，然后用蛋白酶K和氯仿-苯酚混合物进行脱蛋白处理，制备了叶绿体DNA （ct-DNA）。由于获得的ct-DNA没有被三种限制性内切酶消化，因此使用Funakoshi Co.的GENECLEAN商业试剂盒进一步纯化了9个ct-DNA。通过三种限制性内切酶（More Hind III, EcoR I, Sal I）处理检查细胞质dna的变化，然后用1%的脂糖凝胶电泳检测消化产物中每个片段的总数和迁移率。9种植物叶片的ct - dna电泳结果非常相似，但有几个片段带是不同的。特别是在萝卜和萝卜这两个物种之间，两者的电泳模式有明显的差异。十字花科植物的二倍体和四倍体、CMS种和正常种之间没有差异。在杂交植物中，油菜的限制性内切酶酶切模式与萝卜非常相似。最后，我们认为与CMA相关的ct- dna没有变异。从9种十字花科植物主根中纯化线粒体dna （mt-DNA）。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，所以尝试失败了。主根中mt- dna的产量因主根年龄的不同而有很大差异。其中一些太低，无法分析mt- dna的变化。mt- dna的变异可能大于ct- dna，这可能与CMS有关。少

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