Max-Planck-Institut für Züchtungsforschung, Abteilung Pflanzenzüchtung und Ertragsphysiologie, D-50829, Köln, Germany.
Theor Appl Genet. 1996 Mar;92(3-4):455-62. doi: 10.1007/BF00223693.
A successful hybridization of a diploid clone of Solanum tuberosum with a rolC-transgenic, diploid S. papita clone is reported. By using leaf expiants of this S. papita clone, which after transformation expressed kanamycin resistance, intact protoplasts were obtained, but these protoplasts did not develop to microcalli or regenerate to mature plants. However, protoplasts of the S. tuberosum clone showed a high capacity to regenerate plants from isolated protoplasts. On a medium containing Kanamycin only calli regenerated to plants, which revealed a rolC phenotype (reduced apical dominance with a large number of adventitious shoots and a pale green color of leaves) and later on turned out to be true hybrids. Self fusions of S. papita never developed to microcalli and those of S. tuberosum ceased to develop on the kanamycin-containing medium. Identification of somatic hybrids was done by RFLP and RAPD analysis. In the greenhouse, out of four selected hybrids only FK3.1 was successfully crossed with two standard S. tuberosum varieties (Datura, Desirée). Out of all the seeds germinated, only rolC-negative F(1) seedlings were further characterized. Within the seedling population obvious differences were evident in respect of the S. papita and S. tuberosum characteristics.
报道了用 rolC 转基因二倍体 S. papita 克隆与二倍体克隆杂种马铃薯的成功杂交。利用这种 S. papita 克隆的叶片外植体，转化后表达卡那霉素抗性，获得了完整的原生质体，但这些原生质体未能发育成小愈伤组织或再生为成熟植株。然而，马铃薯克隆的原生质体表现出从分离的原生质体再生植物的高能力。在仅含有卡那霉素的培养基上，愈伤组织再生为具有 rolC 表型（顶端优势减弱，产生大量不定芽，叶片呈浅绿色）的植株，后来被证明是真正的杂种。S. papita 的自融合从未发育成小愈伤组织，而 S. tuberosum 的自融合在含卡那霉素的培养基上停止发育。体细胞杂种的鉴定是通过 RFLP 和 RAPD 分析进行的。在温室中，从四个选定的杂种中，只有 FK3.1 成功地与两个标准的马铃薯品种（Datura、Desirée）杂交。在所有发芽的种子中，只有 rolC 阴性的 F(1) 苗进一步进行了特征描述。在幼苗群体中，在 S. papita 和 S. tuberosum 的特征方面明显存在差异。
