Department of Plant Production, University of Helsinki, SF-00710, Helsinki, Finland.
Theor Appl Genet. 1993 Feb;85(8):1017-22. doi: 10.1007/BF00215042.
Mitochondrial (mt) DNAs of somatic hybrids obtained by electrical and chemical fusion of mesophyll protoplasts of S. brevidens and a dihaploid line of S. tuberosum PDH 40 were analysed by Southern hybridization using the digoxigenin-labelled mtDNA sequences nad5 or orf25. In the Southern analysis of the hybrid mtDNA probed with nad5, most of the 19 hybrids analyzed had an RFLP pattern similar, but not identical, to one of the parents, S. tuberosum, PDH40. Nineteen percent of the hybrids had most of the S. brevidens fragments. Five of the hybrids had an identical RFLP pattern to either one of the parents while another two hybrids had novel RFLP patterns. Similar results were obtained by Southern analysis with orf25. These results clearly show that mtDNA rearrangements had occurred at a high frequency in the somatic hybrids. There were no differences in the frequencies of rearrangements observed between the hybrids regenerated from chemical and electrical fusions.
用电融合和化学融合的方法,将短柄野芝麻的叶肉原生质体与二倍体马铃薯 PDH40 杂种细胞融合,得到了体细胞杂种。利用 Southern 杂交,用标记的线粒体 nad5 或 orf25 序列对杂种的线粒体 DNA 进行了分析。用 nad5 探针进行 Southern 杂交分析,在所研究的 19 个杂种中,大多数杂种的 RFLP 图谱与亲本之一的马铃薯 PDH40 相似,但不完全相同。19%的杂种具有大多数短柄野芝麻的片段。有 5 个杂种与亲本之一具有相同的 RFLP 图谱,而另外两个杂种则具有新的 RFLP 图谱。用 orf25 进行 Southern 分析得到了相似的结果。这些结果清楚地表明,线粒体 DNA 重排发生在体细胞杂种中频率很高。在化学融合和电融合再生的杂种中,观察到的重排频率没有差异。