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.

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.

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.

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.

Identification of all chromosome arms and their involvement in meiotic homoeologous associations at metaphase I in 2 Hordeum vulgare L. × Hordeum bulbosum L. hybrids

Genome ◽  
2006 ◽  
Vol 49 (1) ◽  
pp. 73-78 ◽  
Author(s):  
R Pickering ◽  
S Klatte ◽  
R C Butler
Keyword(s):  
In Situ Hybridization ◽  
Hordeum Vulgare ◽  
Fluorescence In Situ Hybridization ◽  
Interspecific Hybrids ◽  
Association Studies ◽  
Hordeum Bulbosum ◽  
Hordeum Vulgare L ◽  
Chromosome Addition ◽  
Chromosome Associations

We have identified all Hordeum bulbosum chromosomes in 2 diploid Hordeum vulgare × Hordeum bulbosum hybrids using suitable probes and fluorescence in situ hybridization. Using the parental idiograms allowed us to carry out a full analysis of chromosome associations among all chromosome arms in the hybrids. Association frequencies were generally lower for the short arms than for the long arms. There were also significant differences among the chromosome arms in association frequencies, partly correlated with the absolute length of the chromosome arm, as well as with the frequency of recombinant lines, which were recovered from partially fertile interspecific hybrids. The H. bulbosum idiogram will be useful for further chromosome association studies and will enable the identification of H. bulbosum chromosomes involved in chromosome addition or substitution lines.Key words: Hordeum vulgare, Hordeum bulbosum, interspecific hybrids, chromosome associations, meiosis, fluorescence in situ hybridization.

scholarly journals Degradabilidad y estimación del consumo de forrajes y concentrados en alpacas (Vicugna pacos)

2018 ◽  
Vol 29 (2) ◽  
pp. 429
Author(s):  
Alfonso Cordero F. ◽  
José Contreras P. ◽  
James Curasma C. ◽  
Miguel Tunque Q. ◽  
Daniel Enríquez Q.
Keyword(s):  
Triticum Aestivum ◽  
Zea Mays ◽  
Solanum Tuberosum ◽  
Hordeum Vulgare ◽  
Avena Sativa ◽  
Zea Mays L ◽  
Triticum Aestivum L ◽  
Hordeum Vulgare L ◽  
Vicugna Pacos

El estudio tuvo como objetivo evaluar los parámetros cinéticos de la degradación in situ de la materia seca (MS), proteína cruda (PC) y la estimación del consumo mediante ecuaciones de predicción de MS de forrajes y alimentos concentrados en alpacas Huacaya (Vicugna pacos). Se trabajó con ensilado de maíz chala (Zea mays L) sin y con 1% de urea, cebada (Hordeum vulgare L), avena (Avena sativa L), salvado de trigo (Triticum aestivum L) y raspa de papa (Solanum tuberosum). Los alimentos (5 g en base seca) fueron colocados en sacos de nylon e incubados en el primer compartimento estomacal de dos alpacas fistuladas durante 0, 6, 12, 24, 48 y 76 horas. Se analizó la MS y la PC de los residuos de los sacos. La MS y la PC del salvado de trigo y de la raspa de papa presentaron potenciales de degradación elevados, así como la MS y la PC de la avena. Se destaca la mayor fracción no degradable de la PC del maíz chala sin y con urea y, por tanto, una menor degradabilidad de la PC. Las estimaciones del consumo por las alpacas generadas por las ecuaciones de tres estudios no son adecuadas a los alimentos en estudio.

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.

High-density fluorescence in situ hybridization signal detection on barley (Hordeum vulgare L.) chromosomes with improved probe screening and reprobing procedures

Genome ◽  
2011 ◽  
Vol 54 (2) ◽  
pp. 151-159 ◽  
Author(s):  
Akio Kato
Keyword(s):  
In Situ Hybridization ◽  
Hordeum Vulgare ◽  
Fluorescence In Situ Hybridization ◽  
Gel Electrophoresis ◽  
Agarose Gel ◽  
Agarose Gel Electrophoresis ◽  
Hordeum Vulgare L ◽  
Pcr Products ◽  
Break Points

The barley ( Hordeum vulgare L.) genome was screened to identify sequences that could be used for fluorescence in situ hybridization (FISH). From 2000 transformed bacterium colonies carrying barley clones, 56 colonies were selected on the basis of the patterns that their PCR products produced when subjected to agarose gel electrophoresis. Among them, 42 (75%) exhibited fluorescent signals on barley chromosomes after in situ hybridization using the directly labeled PCR products. Sequencing revealed seven clones, pHv-365, pHv-177, pHv-1112, pHv-689, pHv-1476, pHv-1889, and pHv-1972, to be newly identified FISH-positive sequences. The remainder possess previously described sequences such as 5S, GAA microsatellite, centromere repeats, HVT01, and pHvMWG2315 (324 bp repeat). It is shown here that a combination of five probes, which produce strong signals on barley chromosomes, pHv-38 (5S), pHv-365, pHv-961 (HVT01), GAA, and TAG microsatellites, offer unequivocal recognition of each chromosome. The combination of three probes, i.e., pHv-1123 (barley 324 bp repeat), GAA, and TAG, decorated entire chromosomes with fine dotted signals and was useful for detecting the break points of aberrant chromosomes. The signals’ distributions of pHv-177, pHv-1112, and TAG were highly polymorphic. An improved reprobing procedure and its usefulness are also discussed.

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