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采用PEG(聚乙二醇)融合法,诱导水稻(Oryza sativa L.)原生质体与无融合生殖大黍(Panicummaximum Jacq.)原生质体融合,经过融合体筛选、培养,成功地获得了再生植株并移栽成活。在融合前,水稻原生质体经过2.5mmol/L碘乙酰胺(IOA)在室温(22～25℃)条件下处理15min,大黍原生质体经过60Kr软X射线照射处理。对获得的28株融合再生植株进行初步检查发现,在花器官形态、结构及生殖特性上与对照亲本水稻植株有显著的差异,出现多花药(一朵颖花具7～11枚甚至13枝花药)、多胚珠(1个子房内有2～3个胚珠)及“多胚囊”(1个胚珠内有2个以上类似胚囊的结构)等现象。雌、雄性育性显著降低或完全不育,仅有5株能够少量结实,I-KI溶液着色的花粉从0至68％不等。细胞胚胎学检查表明不能结实的植株雌性均不育,即不能分化出正常的胚囊结构。
The protoplasts of rice (Oryza sativa L.) were induced to fuse with protoplasts of Panicum maximum Jacq. By PEG (polyethylene glycol) fusion method. The regenerated plants were successfully obtained through fusion selection and culture Transplant survival. Before fusion, protoplasts of rice were treated with 2.5mmol / L iodoacetamide (IOA) for 15min at room temperature (22 ~ 25 ℃). Protoplasts of rice were treated with soft X-ray irradiation of 60Kr. The preliminary examination of the 28 fusion regenerated plants revealed that there were significant differences in flower organ morphology, structure and reproductive characteristics between the parental rice plants and the control parental plants (7 to 11 or even 13 anthers ), Multiple ovules (2-3 ovules in 1 ovary), and “multiple embryo sacs” (two or more embryo sac-like structures in one ovule). Female or male fertility was significantly reduced or completely sterile, and only 5 plants were able to be sturdily small, ranging from 0 to 68% with I-KI solution-colored pollen. Cellular embryological examination showed that plants that can not be sturdy are female sterile, that is, can not differentiate into normal embryo sac structure.