Genome. 1997 Jun;40(3):362-9. doi: 10.1139/g97-049.
Southern and in situ hybridization were used to examine the chromosome constitution, genomic relationships, repetitive DNA sequences, and nuclear architecture in durum wheat x tritordeum hybrids (2n = 5x = 35), where tritordeum is the fertile amphiploid (2n = 6x = 42) between Hordeum chilense and durum wheat. Using in situ hybridization, H. chilense total genomic DNA hybridized strongly to the H. chilense chromosomes and weakly to the wheat chromosomes, which showed some strongly labelled bands. pHcKB6, a cloned repetitive sequence isolated from H. chilense, enabled the unequivocal identification of each H. chilense chromosome at metaphase. Analysis of chromosome disposition in prophase nuclei, using the same probes, showed that the chromosomes of H. chilense origin were in individual domains with only limited intermixing with chromosomes of wheat origin. Six major sites of 18S-26S rDNA genes were detected on the chromosomes of the hybrids. Hybridization to Southern transfers of restriction enzyme digests using genomic DNA showed some variants of tandem repeats, perhaps owing to methylation. Both techniques gave complementary information, extending that available from phenotypic, chromosome morphology, or isozyme analysis, and perhaps are useful for following chromosomes or chromosome segments during further crossing of the lines in plant breeding programs.
采用Southern 杂交和原位杂交技术,对硬粒小麦与中间偃麦草衍生的 6 倍体杂种(2n=5x=35)的染色体组成、基因组关系、重复 DNA 序列和核型结构进行了研究。中间偃麦草是由 Hordeum chilense 和硬粒小麦杂交衍生的可育异源六倍体(2n=6x=42)。利用原位杂交,H. chilense 总基因组 DNA 与 H. chilense 染色体强烈杂交,与小麦染色体微弱杂交,显示出一些强烈标记的带。从 H. chilense 中分离的克隆重复序列 pHcKB6 能够在中期使每个 H. chilense 染色体得以明确识别。使用相同的探针分析前期核中的染色体排列,表明源自 H. chilense 的染色体位于单个结构域中,与源自小麦的染色体仅有有限的混合。在杂种的染色体上检测到 6 个 18S-26S rDNA 基因的主要位点。使用基因组 DNA 对酶切消化的 Southern 转移进行杂交显示出一些串联重复的变体,可能是由于甲基化所致。这两种技术都提供了互补的信息,扩展了表型、染色体形态或同工酶分析提供的信息,并且在植物育种计划中进一步杂交这些系时,可能有助于跟踪染色体或染色体片段。