用于ND-FISH分析以鉴定黑麦和小麦染色体的寡核苷酸探针
Oligonucleotide Probes for ND-FISH Analysis to Identify Rye and Wheat Chromosomes.
作者信息
Fu Shulan, Chen Lei, Wang Yangyang, Li Meng, Yang Zujun, Qiu Ling, Yan Benju, Ren Zhenglong, Tang Zongxiang
机构信息
1] Province Key Laboratory of Plant Breeding and Genetics, Sichuan Agricultural University, Wenjiang, Chengdu 611130, Sichuan, People's Republic of China [2] Agronomy College, Sichuan Agricultural University, Wenjiang, Chengdu 611130, Sichuan, People's Republic of China.
School of Life Science and Technology, University of Electronic Science and Technology of China, Chengdu 610054, Sichuan, People's Republic of China.
出版信息
Sci Rep. 2015 May 21;5:10552. doi: 10.1038/srep10552.
Genomic in situ hybridization (GISH) has been widely used to detect rye (Secale cereale L.) chromosomes in wheat (Triticum aestivum L.) introgression lines. The routine procedure of GISH using genomic DNA of rye as a probe is time-consuming and labor-intensive because of the preparation and labeling of genomic DNA of rye and denaturing of chromosomes and probes. In this study, new oligonucleotide probes Oligo-1162, Oligo-pSc200 and Oligo-pSc250 were developed. The three new probes can be used for non-denaturing fluorescence in situ hybridization (ND-FISH) assays and replace genomic DNA of rye as a probe to discriminate rye chromosomes in wheat backgrounds. In addition, previously developed oligonucleotide probes Oligo-pSc119.2-1, Oligo-pSc119.2-2, Oligo-pTa535-1, Oligo-pTa535-2, Oligo-pTa71-2, Oligo-pAWRC.1 and Oligo-CCS1 can also be used for ND-FISH of wheat and rye. These probes have provided an easier, faster and more cost-effective method for the FISH analysis of wheat and hybrids derived from wheat × rye.
基因组原位杂交（GISH）已被广泛用于检测小麦（Triticum aestivum L.）渐渗系中的黑麦（Secale cereale L.）染色体。由于黑麦基因组DNA的制备、标记以及染色体和探针的变性，以黑麦基因组DNA为探针的常规GISH程序既耗时又费力。在本研究中，开发了新的寡核苷酸探针Oligo-1162、Oligo-pSc200和Oligo-pSc250。这三种新探针可用于非变性荧光原位杂交（ND-FISH）分析，并替代黑麦基因组DNA作为探针，以区分小麦背景中的黑麦染色体。此外，先前开发的寡核苷酸探针Oligo-pSc119.2-1、Oligo-pSc119.2-2、Oligo-pTa535-1、Oligo-pTa535-2、Oligo-pTa71-2、Oligo-pAWRC.1和Oligo-CCS1也可用于小麦和黑麦的ND-FISH。这些探针为小麦及其与黑麦杂交后代的FISH分析提供了一种更简便、快速且经济高效的方法。
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