Oligonucleotide Probes for ND-FISH Analysis to Identify Rye and Wheat Chromosomes

• Published in: Scientific reports Vol. 5; no. 1; p. 10552
• Main Authors: Fu, Shulan, Chen, Lei, Wang, Yangyang, Li, Meng, Yang, Zujun, Qiu, Ling, Yan, Benju, Ren, Zhenglong, Tang, Zongxiang
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 21.05.2015; Nature Publishing Group
• Subjects: 13; 14; 38/32; 631/1647/1407/1405; 631/449/711; Chromosomes; Chromosomes, Plant - genetics; Chromosomes, Plant - metabolism; Deoxyribonucleic acid; DNA; DNA probes; Fluorescence in situ hybridization; Fluorescent indicators; Genomic in situ hybridization; Humanities and Social Sciences; Hybrids; In Situ Hybridization, Fluorescence; Microscopy, Fluorescence; multidisciplinary; Oligonucleotide Probes - genetics; Oligonucleotide Probes - metabolism; Oligonucleotides; Polyploidy; Science; Secale - genetics; Triticum - genetics; Triticum aestivum
• ISSN: 2045-2322; 2045-2322
• 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.