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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.
Highlights
In situ hybridization (ISH) is a molecular, cytogenetic technique that uses labeled DNA or RNA probes to directly target specific nucleic acid sequences on chromosomes [1,2]
The main advantages of non-denaturing fluorescence in situ hybridization (ND-Fluorescence in situ hybridization (FISH)) are as follows: It does not require the preparation of probes which can be synthesized commercially, there is no denaturation process and a short hybridization time, and it has a lower cost than FISH [13,14]
In the 2A chromosomes, the signal from Oligo-pTa535-1 on the 2A long arm was missing in CN21, CN22, CN23, CN25 cultivars, and an additional signal from Oligo-pSc119.2-1 was present on the 2A long arm in CN19, CN26, CN27, CN28, CN29, CN30, CN31, CN32, CN33 and CN35 cultivars (Figure 1)
Summary
In situ hybridization (ISH) is a molecular, cytogenetic technique that uses labeled DNA or RNA probes to directly target specific nucleic acid sequences on chromosomes [1,2]. The main advantages of ND-FISH are as follows: It does not require the preparation of probes which can be synthesized commercially, there is no denaturation process and a short hybridization time, and it has a lower cost than FISH [13,14] Based on these advantages, this technique has been widely used to identify wheat and chromosomes from alien species [9,13,14,15,16]. The design of the oligonucleotide probes is the key to accurately identifying
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