FISH landmarks for barley chromosomes (Hordeum vulgare L.)

Genome ◽  
1999 ◽  
Vol 42 (2) ◽  
pp. 274-281 ◽  
Author(s):  
Susan E Brown ◽  
Janice L Stephens ◽  
Nora LV Lapitan ◽  
Dennis L Knudson
Keyword(s):  
Hordeum Vulgare ◽  
Digital Imaging ◽  
18S Rdna ◽  
Chromosomal Location ◽  
5S Rdna ◽  
Hordeum Vulgare L ◽  
Metaphase Chromosomes ◽  
Bac Clone ◽  
Rdna Probe

Barley metaphase chromosomes (2n = 14) can be identified by fluorescence in situ hybridization (FISH) and digital imaging microscopy using heterologous 18S rDNA and 5S rDNA probe sequences. When these sequences are used together, FISH landmark signals were seen so that all 7 chromosomes were uniquely identified and unambiguously oriented. The chromosomal location of the landmark signals was determined by FISH to a barley trisomic series using the 18S and 5S probes labeled with different fluorophores. The utility of these FISH landmarks for barley physical mapping was also demonstrated when an Amy-2 cDNA clone and a BAC clone were hybridized with the FISH landmark probes.Key words: Hordeum vulgare, barley, FISH, 5S, 18S, rDNA, landmarks, chromosome.

scholarly journals Interindividual size heteromorphism of NOR and chromosomal location of 5S rRNA genes in Iheringichthys labrosus

2007 ◽  
Vol 50 (1) ◽  
pp. 141-146 ◽  
Author(s):  
Rafael Augusto de Carvalho ◽  
Ana Lúcia Dias
Keyword(s):  
Chromosomal Location ◽  
5S Rdna ◽  
Rrna Genes ◽  
Telomeric Region ◽  
Fluorescent Staining ◽  
Homologous Chromosomes ◽  
Rdna Probe ◽  
5S Rrna Genes ◽  
Iheringichthys Labrosus

Twenty-five specimens of Iheringichthys labrosus from the Capivara Reservoir were analysed cytogenetically. AgNORs were detected in a pair of ST chromosomes, in the telomeric region of the long arm. Some individuals showed size heteromorphism of this region between homologous chromosomes. Treatment with CMA3 displayed GC-rich regions corresponding to the AgNOR pair, plus other fluorescent staining. In situ hybridization by fluorescence (FISH) with the 18S rDNA probe showed only one pair of stained chromosomes, confirming the heteromorphism observed with AgNO3 and CMA3 in some individuals. The 5S rDNA probe revealed telomeric staining on the long arm of a pair of chromosomes of the ST-A group, probably different from the NOR pair.

Physical mapping of four sites of 5S rDNA sequences and one site of the α-amylase-2 gene in barley (Hordeum vulgare)

Genome ◽  
1993 ◽  
Vol 36 (3) ◽  
pp. 517-523 ◽  
Author(s):  
I. J. Leitch ◽  
J. S. Heslop-Harrison
Keyword(s):  
In Situ Hybridization ◽  
Hordeum Vulgare ◽  
Physical Mapping ◽  
5S Rdna ◽  
Chromosome 1 ◽  
Chromosome 2 ◽  
Hordeum Vulgare L ◽  
Rdna Sequences ◽  
5S Rdna Sequence

The 5S rDNA sequences have been mapped on four pairs of barley (Hordeum vulgare L.) chromosomes using in situ hybridization and barley monotelotrisomic lines. The 5S rDNA sequences are located, genetically and physically, on the short arm of chromosome 1 (7I) and the long arms of chromosomes 2 (2I) and 3 (3I). The 5S rDNA sequence is also located on the physically long arm of chromosome 4 (4I). Only one site on chromosome 2(2I) has previously been reported. The characteristic locations of the 5S rDNA sequences make them useful as molecular markers to identify each barley chromosome. The physical position of the low-copy α-amylase-2 gene was determined using in situ hybridization; the location of this gene on the long arm of chromosome 1 (7I) was confirmed by reprobing the same preparation with the 5S rDNA probe. The results show that there is a discrepancy between the physical and genetic position of the α-amylase-2 gene.Key words: genetic mapping, physical mapping, barley, mapping, 5S DNA, α-amylase, in situ hybridization.

INHERITANCE AND LINKAGE STUDIES WITH THE GRANDPA GENE IN BARLEY, HORDEUM VULGARE L.

1971 ◽  
Vol 13 (3) ◽  
pp. 489-498
Author(s):  
R. W. Matchett ◽  
H. G. Nass ◽  
D. W. Robertson
Keyword(s):  
Hordeum Vulgare ◽  
Chromosomal Location ◽  
Gene Interactions ◽  
Chromosome 2 ◽  
Linkage Studies ◽  
Hordeum Vulgare L ◽  
Barley Genome ◽  
Chlorophyll Mutants ◽  
Mutant Genes ◽  
Linkage Association

This study was initiated to determine the chromosomal location of the grandpa (gp) gene within the barley genome. The gp gene was placed on the long arm of chromosome 2 as indicated by linkage association with liguleless (li).Tests of allelism showed the gp gene to the allelic with the gp-2 gene. Seven sources of "yellow" chlorophyll mutants when crossed to grandpa plants gave albino double recessive seedlings. Three other sources of "yellow" chlorophyll mutants in the double recessive combination with grandpa exhibited yellow and white bands on the leaves. Double recessive individuals carrying the mottled (mt2) and grandpa genes were also albino. This is evidence of gene interactions between chlorophyll mutant genes.

scholarly journals Brief communication. In situ hybridization mapping of genes in Hordeum vulgare L.

1998 ◽  
Vol 89 (4) ◽  
pp. 366-370 ◽  
Author(s):  
G Butnaru ◽  
J Chen ◽  
P Goicoechea ◽  
JP Gustafson
Keyword(s):  
In Situ Hybridization ◽  
Hordeum Vulgare ◽  
Hordeum Vulgare L ◽  
Hybridization Mapping

Physical mapping of barley genes using an ultrasensitive fluorescence in situ hybridization technique

Genome ◽  
2004 ◽  
Vol 47 (1) ◽  
pp. 179-189 ◽  
Author(s):  
J L Stephens ◽  
S E Brown ◽  
N L.V Lapitan ◽  
D L Knudson
Keyword(s):  
In Situ Hybridization ◽  
Hordeum Vulgare ◽  
Fluorescence In Situ Hybridization ◽  
Linkage Map ◽  
Physical Mapping ◽  
Physical Map ◽  
Primary Objective ◽  
Hybridization Technique ◽  
Hordeum Vulgare L

The primary objective of this study was to elucidate gene organization and to integrate the genetic linkage map for barley (Hordeum vulgare L.) with a physical map using ultrasensitive fluorescence in situ hybridization (FISH) techniques for detecting signals from restriction fragment length polymorphism (RFLP) clones. In the process, a single landmark plasmid, p18S5Shor, was constructed that identified and oriented all seven of the chromosome pairs. Plasmid p18S5Shor was used in all hybridizations. Fourteen cDNA probes selected from the linkage map for barley H. vulgare 'Steptoe' × H. vulgare 'Morex' (Kleinhofs et al. 1993) were mapped using an indirect tyramide signal amplification technique and assigned to a physical location on one or more chromosomes. The haploid barley genome is large and a complete physical map of the genome is not yet available; however, it was possible to integrate the linkage map and the physical locations of these cDNAs. An estimate of the ratio of base pairs to centimorgans was an average of 1.5 Mb/cM in the distal portions of the chromosome arms and 89 Mb/cM near the centromere. Furthermore, while it appears that the current linkage maps are well covered with markers along the length of each arm, the physical map showed that there are large areas of the genome that have yet to be mapped.Key words: Hordeum vulgare, barley, physical mapping, FISH, cDNA, genetics, linkage, chromosome, BACs.

Integrated Karyotyping of Woodland Strawberry (Fragaria vesca) with Oligopaint FISH Probes

2017 ◽  
Vol 153 (3) ◽  
pp. 158-164 ◽  
Author(s):  
Manman Qu ◽  
Kunpeng Li ◽  
Yanli Han ◽  
Lei Chen ◽  
Zongyun Li ◽  
...  
Keyword(s):  
Draft Genome ◽  
5S Rdna ◽  
Fish Analysis ◽  
Fragaria Vesca ◽  
Chromosome Identification ◽  
Multicolor Fish ◽  
Metaphase Chromosomes ◽  
Cross Species Chromosome Painting ◽  
Simultaneous Identification

Chromosome identification is critical for many aspects of cytogenetic research. However, for Fragaria vesca, definite identification of individual chromosomes is almost impossible because of their small size and high similarity. Here, we demonstrate that bulked oligonucleotide (oligo) probes can be used as chromosome-specific DNA markers for chromosome identification in F. vesca. Oligos specific to entire pseudochromosomes in the draft genome of F. vesca were identified and synthesized as libraries. In all, we synthesized 6 oligo libraries corresponding to 6 pseudochromosomes of F. vesca. These libraries were amplified and labeled as probes for fluorescence in situ hybridization (FISH). Two rounds of multicolor FISH analysis were sequentially conducted on the same metaphase cells with each round including 3 probe libraries, which permitted simultaneous identification of all chromosomes of F. vesca. Moreover, 45S and 5S rDNA were mapped to chromosomes 1, 2, and 7, respectively. A karyotype of metaphase chromosomes was constructed, representing the first FISH-based molecular cytogenetic karyotype of F. vesca. Our study can serve as a basis for future comparative cytogenetic research through cross-species chromosome painting using bulked oligo probes and will facilitate the application of breeding technologies that rely on the identification of chromosomes in the genus Fragaria.

Characterization by RFLP analysis and genomic in situ hybridization of a recombinant and a monosomic substitution plant derived from Hordeum vulgare L. × Hordeum bulbosum L. crosses

Genome ◽  
1997 ◽  
Vol 40 (2) ◽  
pp. 195-200 ◽  
Author(s):  
R. A. Pickering ◽  
A. M. Hill ◽  
R. G. Kynast
Keyword(s):  
In Situ Hybridization ◽  
Hordeum Vulgare ◽  
Rflp Analysis ◽  
Leaf Sheath ◽  
Genomic In Situ Hybridization ◽  
Barley Chromosome ◽  
Interspecific Crosses ◽  
Hordeum Bulbosum ◽  
Hordeum Vulgare L

Interspecific crosses in Hordeum have been made with the aim of transferring desirable traits, such as disease resistance, from a wild species, Hordeum bulbosum, into cultivated barley (Hordeum vulgare). Interspecific recombinants have previously been identified using several methods, but there are limitations with all the techniques. We improved our ability to characterize progeny from H. vulgare × H. bulbosum crosses by using genomic in situ hybridization (GISH). The plant material comprised a recombinant and a monosomic alien substitution plant derived from H. vulgare × H. bulbosum crosses. The recombinant possesses a pubescent leaf sheath conferred by a gene transferred from H. bulbosum into barley cultivar Golden Promise. The use of GISH on a plant homozygous for the pubescence gene confirmed the presence of H. bulbosum DNA located distally on two barley chromosomes and we mapped the introgression to barley chromosome 4HL using RFLP analysis. Furthermore, by means of an allelism test we found that the transferred gene for pubescence is allelic or closely linked to a gene for pubescence (Hs) located on barley chromosome 4HL. The presence of a single H. bulbosum chromosome in the monosomic substitution plant was confirmed by GISH. A distal introgression of H. bulbosum DNA was also observed on one barley chromosome, which was located on chromosome 3HL by RFLP analysis.Key words: Hordeum vulgare, Hordeum bulbosum, interspecific hybrid, gene introgression, genomic in situ hybridization.

In situ digestion of barley chromosomes with restriction endonucleases

Genome ◽  
1992 ◽  
Vol 35 (5) ◽  
pp. 793-798 ◽  
Author(s):  
Y. Kamisugi ◽  
Y. Ikeda ◽  
M. Ohno ◽  
M. Minezawa ◽  
K. Fukui
Keyword(s):  
Hordeum Vulgare ◽  
Restriction Endonuclease ◽  
Treatment Time ◽  
Band Pattern ◽  
Restriction Endonucleases ◽  
Chromosome Band ◽  
Buffer Solution ◽  
Hordeum Vulgare L

In situ digestion of barley chromosomes with restriction endonucleases was examined. All the treatments with five restriction endonucleases, MboII, RsaI, HaeIII, HinfI, and DraII, showed various band patterns on the barley chromosomes. Differences were observed in the band patterns produced with different restriction endonucleases. Uneven staining patterns, similar to the band patterns by the endonuclease treatments, also appeared when the chromosomes were treated with the buffer solution without the enzyme. The band patterns observed both with and without the endonucleases were classified into the four types and the frequency of each type among the different treatments was investigated. The change of the band types along with treatment time was accelerated by the addition of the restriction endonuclease. As a result, it was concluded that there existed chromosome band patterns that were specific to the endonuclease treatments and that the buffer solution also affected to the production of the bands on the chromosomes.Key words: Hordeum vulgare L., chromosome band pattern, in situ digestion, restriction endonuclease, restriction banding.

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