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和离核融合技术，在胡萝卜中尝试了细胞质雄性不育与正常可育植株的相互转化。60 kR x射线照射和10 mM碘甲酰胺照射后，用PEG处理细胞质，使供、受体细胞系融合。无论使用何种细胞材料，许多植株都是通过体细胞胚胎发生从融合产物中再生出来的。然而，没有获得雄性不育性改变的植株。在对照培养中，仅用x射线或iaa处理，可能由于互补而获得一些逃逸菌落。因此，在融合治疗前采用更强的处理来抑制游离。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是非常困难的，而从液泡化的非胚性细胞中分离细胞核相对容易。然而，利用非胚性细胞分离的细胞核不适合在细胞核结合后获得植株再生。需要进一步的实验从胚胎细胞中获得细胞核。

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.