Yamagishi H, Glimelius K
Department of Biotechnology, Kyoto Sangyo University, Kamigamo, Kita, 603-8555 Kyoto, Japan.
Plant Cell Rep. 2003 Aug;22(1):52-8. doi: 10.1007/s00299-003-0655-0. Epub 2003 Jun 26.
Somatic hybrids were produced by protoplast fusion between Arabidopsis thaliana ecotype Columbia and a male-sterile radish line MS-Gensuke ( Raphanus sativus) with the Ogura cytoplasm. Forty-one shoots were differentiated from the regenerated calli and established as shoot cultures in vitro. About 20 of these shoots were judged to be hybrids based on growth characteristics and morphology. Molecular analyses of 11 shoots were performed, confirming the hybrid features. Of these 11 shoots, eight were established as rooted plants in the greenhouse. Polymerase chain reaction and randomly amplified polymorphic DNA analyses of the nuclear genomes of all analyzed shoots and plants confirmed that they contained hybrid DNA patterns. Their chromosome numbers also supported the hybrid nature of the plants. Investigations of the organelles in the hybrids revealed that the chloroplast (cp) genome was exclusively represented by radish cpDNA, while the mitochondrial DNA configuration showed a combination of both parental genomes as well as fragments unique to the hybrids. Hybrid plants that flowered were male-sterile independent of the presence of the Ogura CMS-gene orf138.
通过拟南芥生态型哥伦比亚与具有小仓细胞质的雄性不育萝卜系MS - 玄助（Raphanus sativus）之间的原生质体融合产生了体细胞杂种。从再生愈伤组织中分化出41个芽，并在体外建立茎尖培养物。根据生长特性和形态，这些芽中约有20个被判定为杂种。对11个芽进行了分子分析，证实了杂种特征。在这11个芽中，有8个在温室中长成了生根植株。对所有分析的芽和植株的核基因组进行聚合酶链反应和随机扩增多态性DNA分析，证实它们含有杂种DNA模式。它们的染色体数目也支持了植株的杂种性质。对杂种中细胞器的研究表明，叶绿体（cp）基因组完全由萝卜cpDNA代表，而线粒体DNA构型显示出双亲基因组以及杂种特有的片段的组合。开花的杂种植株是雄性不育的，与小仓细胞质雄性不育基因orf138的存在无关。
