Examination of the mitochondrial genome of revertant progeny from S cms maize with cloned S-1 and S-2 hybridization probes.

R J Kemble, R J Mans
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Abstract

We have examined the molecular rearrangement of mitochondrial DNAs in each of several fertile revertants that arose spontaneously from S-type cytoplasmically male-sterile maize. Cloned segments of S-1 and S-2 DNAs (plasmid-like DNAs characteristic of the mitochondrial DNAs of S-type lines) were hybridized to untreated and restriction endonuclease-treated mitochondrial DNAs from fertile plants carrying normal cytoplasm, from cytoplasmically male-sterile plants, and from plants that had cytoplasmically reverted to fertility. The relative intensity of hybridization with S-1 and S-2 probes was different among the fertile, sterile, and revertant lines. The sizes of some restriction endonuclease fragments from the fertile revertant lines that hybridize with the S-2 probe differ from those of the sterile parental lines. Preferential synthesis of high molecular weight components of the mitochondrial genome carrying S-1 and S-2 sequences, concomitant with cessation in apparent autonomous replication of discrete S-1 and S-2 DNAs and their replicative intermediates (described here), could accommodate the hybridization data. The results suggest but do not prove that S-2 sequences are transposed coincident with the sterile plant's reversion to fertility. The inserted segments could arise from sequences already present in the high molecular weight DNA or from the lower molecular weight linear components of the mitochondrial genome. Putative target sites of insertion of S-1 and S-2 sequences would be multiple and separate for each. Reversion of S-type cytoplasmically male-sterile plants to fertility does not restore the organization of the mitochondrial genome to that of a normal fertile plant.

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用克隆S-1和S-2杂交探针检测S- cms玉米逆转录子代线粒体基因组。
我们研究了从s型细胞质雄性不育玉米中自发产生的几种可育回复性玉米中线粒体dna的分子重排。将克隆的S-1和S-2 dna片段(s型系线粒体dna的质粒样dna特征)与携带正常细胞质的可育植物、细胞质雄性不育植物和细胞质恢复可育植物的未经处理和限制性内切酶处理的线粒体dna进行杂交。在可育系、不育系和抗性系中,S-1和S-2探针的相对杂交强度不同。与S-2探针杂交的可育逆转录系的一些限制性内切酶片段的大小与不育亲本不同。优先合成携带S-1和S-2序列的线粒体基因组的高分子量组分,同时停止离散S-1和S-2 dna及其复制中间体的明显自主复制(在这里描述),可以容纳杂交数据。结果提示但不能证明S-2序列的转置与不育植株的育性恢复一致。插入的片段可能来自已经存在于高分子量DNA中的序列,也可能来自线粒体基因组中较低分子量的线性成分。假设的S-1和S-2序列的插入靶位是多个且彼此分离的。s型细胞质雄性不育植物恢复可育性并不能使线粒体基因组的组织恢复到正常可育植物的组织。
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