Analysis of genetic differentiations in radish based on the relationship between make sterile cytoplasma and fertility restorer genes

1. Ogura细胞质雄性不育基因orf138的分布和多态性：利用聚合酶链反应技术研究了orf138的分布。orf138在萝卜属以外的其它十字花科植物中未观察到。萝卜属的野生萝卜和日本野萝卜广泛含有该基因，而大多数栽培萝卜缺乏该基因。在143株植物中发现了7个突变，其中6个是核苷酸改变，1个是39bp的缺失。通过组合突变，发现了9种类型的orf 138序列，其中两种主要类型分布在萝卜和日本野萝卜中。这些结果表明orf138基因起源于萝卜，并已遗传到日本野萝卜中.恢复基因的分布与遗传：通过测交，发现不育细胞质的育性恢复基因在野生萝卜中普遍分布。在栽培萝卜中，大多数日本栽培品种不具有该基因，而欧洲和中国栽培品种具有该基因，这一事实表明栽培萝卜的起源是多样的。欧洲栽培萝卜和日本野萝卜的恢复基因均为显性恢复基因，位于同一位点.恢复基因的分子标记：为了找到与在欧洲栽培品种中发现的恢复基因连锁的分子标记，将混合分离分析应用于分离雄性不育的F2群体。筛选出7个RAPD标记，其中1个（OPH11210）与该基因紧密连锁。少数RaPD标记被转化为SCAR标记，这些标记易于检测且稳定。

1. Distribution and polymorphisms of the Ogura cytoplasmic male sterile gene, orf138 : The distribution of orf138 was studies using polymerase chain reaction. The orf138 was not observed in other Cruciferous plants than Raphanus. Wild radishes belonging to R.raphanistrum and Japanese wild radish contain this gene widely, while most of cultivated radishes lack it. For radishes having the oft138, nucleotide sequences of the gene were determined. Seven mutations, six were nucleotide changes and one was a deletion of 39bp, were found in 143 plants. By combination of the mutations, 9 types of the orf 138 sequence were found, among them two major types being distributed in both R.raphanistrum and Japanese wild radish. These results clarifies that the orf138 was originated in R.raphanistrum and was transmitted to Japanese wild radish.2. Distribution and inheritance of the restorer gene : By test-crosses, the fertility restorer gene for the male sterile cytoplasm was found to be distributed generally in wild radishes. In cultivated radishes, most of Japanese cultivars do not have the gene, while European and Chinese cultivars possess it. This fact demonstrates diversified origins of cultivated radishes. The restorer genes found in a European cultivar, R.raphanistrum and a Japanese wild radish are all dominant one locating on an identical locus.3. Molecular Marker of the restorer gene : To find molecular markers linked to the restorer gene found in a European cultivar, bulked segregant analysis was applied to a F2 population segregating for the male sterility. Seven RAPD markers were identified, and one of them (OPH11210) was linked tightly to the gene. A few RaPD markers were converted into SCAR markers which are detectable easily and stably.