Departamento de Genética, Universidad Complutense, Madrid, Spain.
Genome. 2011 Jul;54(7):555-64. doi: 10.1139/g11-003. Epub 2011 Jul 13.
We used rye-specific repetitive DNA sequences in fluorescence in situ hybridization (FISH) to paint the rye genome and to identify rye DNA in a wheat background. A 592 bp fragment from the rye-specific dispersed repetitive family R173 (named UCM600) was cloned and used as a FISH probe. UCM600 is dispersed over the seven rye chromosomes, being absent from the pericentromeric and subtelomeric regions. A similar pattern of distribution was also observed on the rye B chromosomes, but with weaker signals. The FISH hybridization patterns using UCM600 as probe were comparable with those obtained with the genomic in situ hybridization (GISH) procedure. There were, however, sharper signals and less background with FISH. UCM600 was combined with the rye-specific sequences Bilby and pSc200 to obtain a more complete painting. With these probes, the rye chromosomes were labeled with distinctive patterns; thus, allowing the rye cultivar 'Imperial' to be karyotyped. It was also possible to distinguish rye chromosomes in triticale and alien rye chromatin in wheat-rye addition and translocation lines. The distribution of UCM600 was similar in cultivated rye and in the wild Secale species Secale vavilovii Grossh., Secale sylvestre Host, and Secale africanum Stapf. Thus, UCM600 can be used to detect Secale DNA introgressed from wild species in a wheat background.
我们使用黑麦特异性重复 DNA 序列进行荧光原位杂交（FISH），以绘制黑麦基因组并鉴定小麦背景中的黑麦 DNA。克隆了来自黑麦特异性分散重复家族 R173 的 592 bp 片段（命名为 UCM600），并将其用作 FISH 探针。UCM600 分散在七个黑麦染色体上，不存在于着丝粒和端粒区域。在黑麦 B 染色体上也观察到了类似的分布模式，但信号较弱。使用 UCM600 作为探针的 FISH 杂交模式与基因组原位杂交（GISH）程序获得的模式相当。然而，FISH 具有更清晰的信号和更少的背景。UCM600 与黑麦特异性序列 Bilby 和 pSc200 结合使用，以获得更完整的绘制。使用这些探针，黑麦染色体被标记出独特的图案，从而使黑麦品种“Imperial”得以进行核型分析。还可以区分三叶草和小麦-黑麦添加和易位系中的异源黑麦染色质中的黑麦染色体。UCM600 的分布在栽培黑麦和野生黑麦物种 Secale vavilovii Grossh.、Secale sylvestre Host 和 Secale africanum Stapf 中相似。因此，UCM600 可用于检测小麦背景中来自野生种的黑麦 DNA 渐渗。
