Production of cytoplasmic male sterile and its maintainer by using asymmetrical protoplast fusion and incorporation of isolated nuclei.

利用不对称原生质体融合技术，将细胞质雄性不育性导入正常可育植株，并将细胞质雄性不育性导入正常可育植株，将细胞质雄性不育的供体细胞系和受体细胞系分别经60kR X射线照射和10mM碘乙酰胺处理后，用PEG处理进行融合。无论使用的细胞材料，许多植株再生融合产物通过体细胞胚胎发生。然而，没有获得具有改变的雄性不育性的植物。在仅用X射线或IOA处理的对照培养物中，可能由于互补而获得一些逃逸集落。因此，在融合处理之前，应用更强的处理来抑制逃逸。这些处理减少了最终发育成胚胎的存活菌落的数量。进一步的实验正在进行中，从胚性细胞培养物中分离细胞核非常困难，而从空泡化的非胚性细胞中分离细胞核相对容易。然而，使用从非胚性细胞分离的细胞核不适合于在细胞核掺入后获得植株再生。从胚性细胞中获得细胞核还需要进一步的实验。

Introduction of cytoplasmic male sterility into normal fertile plants and vice versa were tried by using asymmetrical protoplast fusion technique and incorporation of isolated nuclei in carrot.Both donor and recipient cell lines of cytoplasm were fused by PEG treatment after irradiation with 60 kR x-ray and treatment with 10 mM iodoactamide, respectively. Irrespective of the cell materials used, many plantlets were regenerated from the fusion products via somatic embryogenesis. However, no plants with altered male sterility were obtained. In the control cultures treated with only x-ray or with IOA, some escaped colonies were obtained presumably due to the complementation. Therefore, stronger treatments were applied to suppress the escape before fusion treatments. These treatments reduced the number of survived colonies which eventually developed into embryos. Further experiments are now in progress.Isolation of nuclei from the embryogenic cell cultures was extremely difficult, whereas that from vacuolated non-embryogenic cells was relatively easy. However, the use of nuclei isolated from non-embryogenic cells was not suitable for obtaining plant regeneration after nuclei incorporation. Further experiment is required to get nuclei from embryogenic cells.