Tang Zongxiang, Yang Zujun, Fu Shulan
State Key Laboratory of Plant Breeding and Genetics, Sichuan Agricultural University, Wenjiang, Chengdu, 611130, Sichuan, People's Republic of China.
J Appl Genet. 2014 Aug;55(3):313-8. doi: 10.1007/s13353-014-0215-z. Epub 2014 Apr 30.
Hybrids derived from wheat (Triticum aestivum L.) × rye (Secale cereale L.) have been widely studied because of their important roles in wheat cultivar improvement. Repetitive sequences pAs1, pSc119.2, pTa-535, pTa71, CCS1, and pAWRC.1 are usually used as probes in fluorescence in situ hybridization (FISH) analysis of wheat, rye, and hybrids derived from wheat × rye. Usually, some of these repetitive sequences for FISH analysis were needed to be amplified from a bacterial plasmid, extracted from bacterial cells, and labeled by nick translation. Therefore, the conventional procedure of probe preparation using these repetitive sequences is time-consuming and labor-intensive. In this study, some appropriate oligonucleotide probes have been developed which can replace the roles of repetitive sequences pAs1, pSc119.2, pTa-535, pTa71, CCS1, and pAWRC.1 in FISH analysis of wheat, rye, and hybrids derived from wheat × rye. These oligonucleotides can be synthesized easily and cheaply. Therefore, FISH analysis of wheat and hybrids derived from wheat × rye using these oligonucleotide probes becomes easier and more economical.
源自小麦(Triticum aestivum L.)×黑麦(Secale cereale L.)的杂种因其在小麦品种改良中的重要作用而受到广泛研究。重复序列pAs1、pSc119.2、pTa - 535、pTa71、CCS1和pAWRC.1通常用作小麦、黑麦以及源自小麦×黑麦的杂种的荧光原位杂交(FISH)分析中的探针。通常,这些用于FISH分析的重复序列中的一些需要从细菌质粒中扩增、从细菌细胞中提取并通过缺口平移进行标记。因此,使用这些重复序列制备探针的传统方法既耗时又费力。在本研究中,已经开发出了一些合适的寡核苷酸探针,它们可以在小麦、黑麦以及源自小麦×黑麦的杂种的FISH分析中取代重复序列pAs1、pSc119.2、pTa - 535、pTa71、CCS1和pAWRC.1的作用。这些寡核苷酸可以轻松且廉价地合成。因此,使用这些寡核苷酸探针进行小麦以及源自小麦×黑麦的杂种的FISH分析变得更加容易且经济。