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 |
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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. |
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ISSN: | 2045-2322 2045-2322 |
DOI: | 10.1038/srep10552 |