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

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 labeli...

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Veröffentlicht in:Scientific reports 2015-05, Vol.5 (1), p.10552-10552, Article 10552
Hauptverfasser: Fu, Shulan, Chen, Lei, Wang, Yangyang, Li, Meng, Yang, Zujun, Qiu, Ling, Yan, Benju, Ren, Zhenglong, Tang, Zongxiang
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Sprache:eng
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Zusammenfassung: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.
ISSN:2045-2322
2045-2322
DOI:10.1038/srep10552