Abstract
Oligonucleotide probes that can be used for non-denaturing fluorescence in situ hybridization (ND-FISH) analysis are convenient tools for identifying chromosomes of wheat (Triticum aestivum L.) and its relatives. New oligonucleotide probes, Oligo-HvT01, Oligo-pTa71-1, Oligo-s120.1, Oligo-s120.2, Oligo-s120.3, Oligo-275.1, Oligo-275.2, Oligo-k566 and Oligo-713, were designed based on the repetitive sequences HVT01, pTa71, pTa-s120, pTa-275, pTa-k566 and pTa-713. All these probes can be used for ND-FISH analysis and some of them can be used to detect polymorphisms of wheat chromosomes. Probes Oligo-HvT01, Oligo-pTa71-1, Oligo-s120.3, Oligo-275.1, Oligo-k566 and Oligo-713 can, respectively, replace the roles of their original sequences to identify chromosomes of some barley (Hordeum vulgare ssp. vulgare) and the common wheat variety Chinese Spring. Oligo-s120.1, Oligo-s120.2 and Oligo-275.2 produced different hybridization patterns from the ones generated by their original sequences. In addition, Oligo-s120.1, Oligo-s120.2 and Oligo-s120.3, which were derived from pTa-s120, revealed different signal patterns. Likewise, Oligo-275.1 and Oligo-275.2, which were derived from pTa-275, also displayed different hybridization patterns. These results imply that differently arranged or altered structural statuses of tandem repeats might exist on different chromosome regions. These new oligonucleotide probes provide extra convenience for identifying some wheat and barley chromosomes, and they can display polymorphisms of wheat chromosomes.
Highlights
Fluorescence in situ hybridization (FISH) techniques have played an important role in modern molecular cytogenetics because they provide an useful tool for chromosome identification, which is an essential prerequisite for deducing the functions of chromosomes
Namely Chinese Spring (CS), Mianyang 11 (MY11), and Chuannong 27 (CN27) as well as three barley varieties (Hordeum vulgare ssp. vulgare), CNSimai 1, PI 328463 and PI 447312, were used to test the new oligonucleotide probes developed in this study
The signal sites of probes pMD-HvT01 and Oligo-HvT01 on the chromosomes of the three kinds of barley were same (Figure S10). These results indicate that the oligonucleotide probes Oligo-HvT01 and Oligo-pTa71-1 can respectively replace the roles of HVT01 and pTa71 to identify chromosomes of some barley varieties
Summary
Fluorescence in situ hybridization (FISH) techniques have played an important role in modern molecular cytogenetics because they provide an useful tool for chromosome identification, which is an essential prerequisite for deducing the functions of chromosomes. Repetitive DNA sequences are often used as probes for FISH analysis on plant chromosomes because they can generate specific FISH signal patterns on individual chromosomes within a single species [1]. Tandem repeats including pAs1, pSc119.2, pTa71 and (AAG)n are mainly used probes in FISH analysis of wheat (Triticum aestivum L.) and its relatives [2,3,4,5,6]. Some new tandemly repeated sequences, including pTa-s120, pTa-275, pTa-535, pTa-k566, pTa-713, etc., have been used as FISH probes to analyze wheat chromosomes [7]. These new repeated sequences are especially valuable probes for FISH analysis of wheat because they
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