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. 2019 Mar 21;24(6):1126.
doi: 10.3390/molecules24061126.

The Polymorphisms of Oligonucleotide Probes in Wheat Cultivars Determined by ND-FISH

Tianheng Ren  1   2 , Maojie He  3   4 , Zixin Sun  5   6 , Feiquan Tan  7   8 , Peigao Luo  9   10 , Zongxiang Tang  11   12 , Shulan Fu  13   14 , Benju Yan  15   16 , Zhenglong Ren  17   18 , Zhi Li  19   20
Affiliations

The Polymorphisms of Oligonucleotide Probes in Wheat Cultivars Determined by ND-FISH

Tianheng Ren et al. Molecules. .

Abstract

Non-denaturing fluorescence in situ hybridization (ND-FISH) has been used to distinguish wheat chromosomes and to detect alien chromosomes in the wheat genome. In this study, five different oligonucleotide probes were used with ND-FISH to examine 21 wheat cultivars and lines. These oligonucleotide probes distinguished 42 wheat chromosomes and also detected rye chromatin in the wheat genome. Moreover, the signal patterns of the oligonucleotide probes Oligo-pTa535-1 and Oligo-pSc119.2-1 showed high polymorphism in the wheat chromosomes. A total of 17.6% of the A group chromosomes, 25.9% of the B group chromosomes and 8.9% of the D group chromosomes showed obvious mutations when they were compared to the standard ND-FISH signal patterns, and most of them were Oligo-pSc119.2-1 mutants. The results suggested that these polymorphisms could be induced by the crossing of wheat cultivars. The results provided more information for the further application of oligonucleotide probes and ND-FISH.

Keywords: ND-FISH; Oligo-pSc119.2-1; Oligo-pTa535-1; chromosome; mutant; wheat.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
The ND-FISH (non-denaturing fluorescence in situ hybridization) results of A-genome chromosomes of all materials. From left to right: CN10, CN11, CN12, CN17, CN18, CN19, CN20, CN21, CN22, CN23, CN24, CN25, CN26, CN27, CN28, CN29, CN30, CN31, CN32, CN33 and CN35, respectively. From top to bottom: 1A, 2A, 3A, 4A, 5A, 6A and 7A chromosome, respectively. The red arrows showed the mutant signal patterns on chromosomes. Yellow: Oligo-pTa535-1; Green: Oligo-pSc119.2-1; Red: Oligo-Ku, Oligo-pSc200 and Oligo-pSc250; Blue: DAPI.
Figure 2
Figure 2
The ND-FISH results of B-genome chromosomes of all materials. From left to right: CN10, CN11, CN12, CN17, CN18, CN19, CN20, CN21, CN22, CN23, CN24, CN25, CN26, CN27, CN28, CN29, CN30, CN31, CN32, CN33 and CN35, respectively. From top to bottom: 1B, 2B, 3B, 4B, 5B, 6B and 7B chromosome, respectively. The red arrows showed the mutant signal patterns on chromosomes. Yellow: Oligo-pTa535-1; Green: Oligo-pSc119.2-1; Red: Oligo-Ku, Oligo-pSc200 and Oligo-pSc250.Blue: DAPI.
Figure 3
Figure 3
The ND-FISH results of D-genome chromosomes of all materials. From left to right: CN10, CN11, CN12, CN17, CN18, CN19, CN20, CN21, CN22, CN23, CN24, CN25, CN26, CN27, CN28, CN29, CN30, CN31, CN32, CN33 and CN35, respectively. From top to bottom: 1D, 2D, 3D, 4D, 5D, 6D and 7D chromosome, respectively. The red arrows showed the mutant signal patterns on chromosomes. Yellow: Oligo-pTa535-1; Green: Oligo-pSc119.2-1; Red: Oligo-Ku, Oligo-pSc200 and Oligo-pSc250.Blue: DAPI.
Figure 4
Figure 4
The differences of ND-FISH signal patterns in CN12 and CN17 cultivars when they were compared with their parents. From left to right: A302, 91S-23, CN12, CN17 and MY11, respectively. The red arrows showed the mutant signal patterns on chromosomes. Yellow: Oligo-pTa535-1; Green: Oligo-pSc119.2-1; Red: Oligo-Ku, Oligo-pSc200 and Oligo-pSc250. Blue: DAPI.
Figure 5
Figure 5
The differences of ND-FISH signal patterns in CN26, CN27 and CN28 cultivars when they were compared with their parents. From left to right: CN19, R3301, CN26, CN27, CN28 and MY11, respectively. The red arrows showed the mutant signal patterns on chromosomes. Yellow: Oligo-pTa535-1; Green: Oligo-pSc119.2-1; Red: Oligo-Ku, Oligo-pSc200 and Oligo-pSc250. Blue: DAPI.
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