Physical organization of repetitive sequences and chromosome diversity of barley revealed by fluorescence in situ hybridization (FISH)

Genome ◽  
2019 ◽  
Vol 62 (5) ◽  
pp. 329-339 ◽  
Author(s):  
Siyu Zhang ◽  
Minqiu Zhu ◽  
Yi Shang ◽  
Jiaqi Wang ◽  
Dawadundup ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Polymorphic Information Content ◽  
Repetitive Sequences ◽  
Chromosome Research ◽  
Sequence Composition ◽  
Induced Mutants ◽  
Chromosome 4H ◽  
Chromosomal Polymorphisms

Fluorescence in situ hybridization (FISH) using oligonucleotides is a simple and convenient method for chromosome research. In this study, 34 of 46 previously developed oligonucleotides produced signals in barley. Together with two plasmid clones and one PCR-amplified cereal centromere repeat (CCS1) probe, 37 repetitive sequences were chromosomally located produced three types of signals covering different positions on the chromosomes. The centromeric and pericentric regions had a more complex genomic organization and sequence composition probably indicative of higher contents of heterochromatin. An efficient multi-plex probe containing eight oligonucleotides and a plasmid clone of 45S rDNA was developed. Thirty-three barley karyotypes were developed and compared. Among them, 11 irradiation-induced mutants of cultivar 08-49 showed no chromosomal variation, whereas 22 cultivar and landrace accessions contained 28 chromosomal polymorphisms. Chromosome 4H was the most variable and 6H was the least variable based on chromosome polymorphic information content (CPIC). Five polymorphic chromosomes (1H-2, 2H-1, 3H-3, 5H-2, and 6H-2) were dominant types, each occurring in more than 50% of accessions. The multi-plex probe should facilitate identification of further chromosomal polymorphisms in barley.

Isolation and characterization of a satellite DNA family in the Saccharum complex

Genome ◽  
1998 ◽  
Vol 41 (6) ◽  
pp. 854-864 ◽  
Author(s):  
Karine Alix ◽  
Franc-Christophe Baurens ◽  
Florence Paulet ◽  
Jean-Christophe Glaszmann ◽  
Angélique D'Hont
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Satellite Dna ◽  
Repetitive Sequences ◽  
Repeat Sequence ◽  
Miscanthus Sinensis ◽  
Saccharum Complex ◽  
Satellite Dnas ◽  
Isolation And Characterization

EaCIR1, a 371-bp Erianthus-specific satellite DNA sequence, was cloned from TaqI restricted genomic DNA after agarose-gel electrophoresis. This sequence has 77% homology with a 365-bp satellite of Helictotrichon convolutum and 72% homology with a 353-bp tandem repeat sequence from Oryza sativa. PCR primers defined in the conserved regions of these repetitive sequences were used to isolate other satellite DNAs in different representatives of the Saccharum complex: SoCIR1 in Saccharum officinarum, SrCIR1 in Saccharum robustum, SsCIR1 and SsCIR2 in Saccharum spontaneum, and MsCIR1 in Miscanthus sinensis. EaCIR1 and SoCIR1 were localized to subtelomeric regions of the chromosomes by fluorescence in situ hybridization. Southern hybridization experiments, using two representatives of this repeat sequence family as probes, illustrated contrasting species-specificity and demonstrated the existence of similar repetitive elements in sorghum and maize.Key words: satellite DNA, sugarcane, Saccharum complex, Gramineae, fluorescence in situ hybridization, FISH.

scholarly journals Comparative Fluorescence in Situ Hybridization Mapping of a 431-kb Arabidopsis thaliana Bacterial Artificial Chromosome Contig Reveals the Role of Chromosomal Duplications in the Expansion of the Brassica rapa Genome

Genetics ◽  
2000 ◽  
Vol 156 (2) ◽  
pp. 833-838 ◽  
Author(s):  
Scott A Jackson ◽  
Zhukuan Cheng ◽  
Ming Li Wang ◽  
Howard M Goodman ◽  
Jiming Jiang
Keyword(s):  
Arabidopsis Thaliana ◽  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Brassica Rapa ◽  
Physical Mapping ◽  
Large Scale ◽  
Repetitive Sequences ◽  
Comparative Genome ◽  
Artificial Chromosomes

Abstract Comparative genome studies are important contributors to our understanding of genome evolution. Most comparative genome studies in plants have been based on genetic mapping of homologous DNA loci in different genomes. Large-scale comparative physical mapping has been hindered by the lack of efficient and affordable techniques. We report here the adaptation of fluorescence in situ hybridization (FISH) techniques for comparative physical mapping between Arabidopsis thaliana and Brassica rapa. A set of six bacterial artificial chromosomes (BACs) representing a 431-kb contiguous region of chromosome 2 of A. thaliana was mapped on both chromosomes and DNA fibers of B. rapa. This DNA fragment has a single location in the A. thaliana genome, but hybridized to four to six B. rapa chromosomes, indicating multiple duplications in the B. rapa genome. The sizes of the fiber-FISH signals from the same BACs were not longer in B. rapa than those in A. thaliana, suggesting that this genomic region is duplicated but not expanded in the B. rapa genome. The comparative fiber-FISH mapping results support that chromosomal duplications, rather than regional expansion due to accumulation of repetitive sequences in the intergenic regions, played the major role in the evolution of the B. rapa genome.

Highly repetitive sequences in B chromosomes of Secale cereale revealed by fluorescence in situ hybridization

Genome ◽  
1994 ◽  
Vol 37 (4) ◽  
pp. 709-712 ◽  
Author(s):  
Angeles Cuadrado ◽  
Nicolas Jouve
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Secale Cereale ◽  
Fluorescent In Situ Hybridization ◽  
Repetitive Sequences ◽  
5S Rdna ◽  
B Chromosomes ◽  
Multigene Families ◽  
Hybridization Probes

An analysis of the presence and distribution of the rye and wheat repeated sequences in rye B chromosomes was carried out by fluorescent in situ hybridization. Probes used consisted of three highly repetitive sequences from rye (pSc119.2, pSc74, and pSc34) and the multigene families for the 25S–5.8S–18S and 5S rDNA from wheat (pTa71 and pTa794, respectively). pSc74 and pSc119.2 showed hybridization signals in the telomeric regions of rye B chromosomes. The remaining DNA clones did not hybridize to the B chromosomes.Key words: Secale cereale, rye, repetitive DNA, fluorescence in situ hybridization, B chromosomes.

scholarly journals New ND-FISH-Positive Oligo Probes for Identifying Thinopyrum Chromosomes in Wheat Backgrounds

2019 ◽  
Vol 20 (8) ◽  
pp. 2031 ◽  
Author(s):  
Wei Xi ◽  
Zongxiang Tang ◽  
Shuyao Tang ◽  
Zujun Yang ◽  
Jie Luo ◽  
...  
Keyword(s):  
Triticum Aestivum ◽  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Repetitive Sequences ◽  
Triticum Aestivum L ◽  
Wheat Breeding ◽  
Fish Analysis ◽  
Breeding Programs ◽  
Gish And Fish

Thinopyrum has been widely used to improve wheat (Triticum aestivum L.) cultivars. Non-denaturing fluorescence in situ hybridization (ND-FISH) technology using oligonucleotides (oligo) as probes provides a convenient and efficient way to identify alien chromosomes in wheat backgrounds. However, suitable ND-FISH-positive oligo probes for distinguishing Thinopyrum chromosomes from wheat are lacking. Two oligo probes, Oligo-B11 and Oligo-pThp3.93, were designed according to the published Thinopyrum ponticum (Th. ponticum)-specific repetitive sequences. Both Oligo-B11 and Oligo-pThp3.93 can be used for ND-FISH analysis and can replace conventional GISH and FISH to discriminate some chromosomes of Th. elongatum, Th. intermedium, and Th. ponticum in wheat backgrounds. The two oligo probes provide a convenient way for the utilization of Thinopyrum germplasms in future wheat breeding programs.

Development and Application of Transposable Element-Based Chromosomal Markers for the St Genome in Triticeae

2019 ◽  
Vol 159 (4) ◽  
pp. 215-224
Author(s):  
Ruijuan Liu ◽  
Feng Yu ◽  
Linna Wei ◽  
Bo Liu ◽  
Demei Liu ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Genomic Dna ◽  
Dna Analysis ◽  
Repetitive Sequences ◽  
Distribution Patterns ◽  
Signal Distribution ◽  
Sequence Composition ◽  
Plasmid Library ◽  
Chromosomal Markers

The St genome, originating from Pseudoroegneria (Nevski) Á. Löve, plays an important role in Triticeae. In this study, the Pseudoroegneria stipifolia genome (2n = 2x = 14, StSt) was screened to identify sequences that could be used for FISH. A total of 163 effective clones were obtained from the genomic plasmid library which was constructed by DNase I digestion of P. stipifolia nuclear genomic DNA. Analysis of these clones identified 112 with characteristics of transposable elements (TEs), 13 with characteristics of tandem repetitive sequences, 8 with characteristics of mRNA sequences, and 30 unknown sequences. Fluorescent signals were detected for 11 of 41 TE sequences on P. stipifolia chromosomes after in situ hybridization and were divided into 4 types according to signal distribution patterns: over the whole St genome chromosomes, telomere to pericentromeric regions, centromere to pericentromeric regions, and terminal regions. The affinity between St and Y genomes was studied using the 11 TE probes in 3 StStYY species. Five TE probes showed no obvious difference between subgenomes, 2 probes displayed divergence only in 2 StStYY species, and 4 probes exhibited significant differences among 3 StStYY species. These results provide a preliminary understanding of the sequence composition of the St genome and enabled 11 novel TE probes to be developed and applied.

Fluorescence in situ hybridization of bovine Alu-like sequences to bovine and ovine chromosomes

Genome ◽  
1993 ◽  
Vol 36 (5) ◽  
pp. 984-986 ◽  
Author(s):  
E. Rajcan-Separovic ◽  
M. P. Sabour
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Repetitive Sequences ◽  
Chromosome Identification ◽  
Banding Patterns ◽  
Alu Sequences ◽  
Diffuse Distribution

Fluorescence in situ hybridization procedures have been applied to study the distribution of Alu-like sequences on bovine and ovine chromosomes. Unlike in man and mouse, where the Alu sequences produced discrete R-like bands, a more diffuse distribution of Alu-like sequences was observed on both bovine and ovine chromosomes. Under the conditions used, banding patterns useful for chromosome identification were not detected.Key words: bovine Alu-like repetitive sequences, bovine and ovine chromosomes, fluorescence in situ hybridization.

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