Detection of barley chromatin added to wheat by genomic in situ hybridization

Genome ◽  
1991 ◽  
Vol 34 (3) ◽  
pp. 448-452 ◽  
Author(s):  
Y. Mukai ◽  
B. S. Gill
Keyword(s):  
In Situ Hybridization ◽  
Genomic Dna ◽  
Repeat Sequence ◽  
Genomic Clone ◽  
Genomic In Situ Hybridization ◽  
Addition Lines ◽  
Interphase Nuclei ◽  
Nucleolar Organizing Region ◽  
Chromosome Domains

A technique for in situ hybridization is reported that can be used to detect barley chromatin in wheat background using total genomic DNA as a probe. A 1:2 ratio of biotin-labeled genomic DNA of barley to blocking (unlabeled, sheared) DNA of wheat was sufficient to reveal brownish labeled barley chromosome domains against bluish background of unlabeled wheat chromatin in metaphase, prophase, and interphase nuclei of wheat-barley addition lines. Using this procedure, the behavior of specific barley chromosomes was analyzed in interphase and prophase cells. In prophase cells, the 6H chromosome was always associated with a nucleolus. A genomic clone of α-amylase gene (gRAmy56) that contains a barley-specific dispersed repeat sequence was also used to detect barley chromosomes in a wheat background.Key words: Hordeum vulgare, Triticum aestivum, genomic in situ hybridization, biotin, nucleolar organizing region.

Genomic in situ hybridization analysis of Thinopyrum chromatin in a wheat - Th. intermedium partial amphiploid and six derived chromosome addition lines

Genome ◽  
1999 ◽  
Vol 42 (6) ◽  
pp. 1217-1223 ◽  
Author(s):  
Qin Chen ◽  
R L Conner ◽  
A Laroche ◽  
W Q Ji ◽  
K C Armstrong ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Genomic Dna ◽  
Genomic In Situ Hybridization ◽  
Dna Probe ◽  
Addition Lines ◽  
Thinopyrum Intermedium ◽  
Chromosome Addition ◽  
Chromosome Addition Lines ◽  
Partial Amphiploid

The genomic origin of alien chromosomes present in a wheat - Thinopyrum intermedium partial amphiploid TAF46 (2n = 8x = 56) and six derived chromosome addition lines were analyzed by genomic in situ hybridization (GISH) using S genomic DNA from Pseudoroegneria strigosa (2n = 2x = 14, SS) as a probe. The GISH analysis clearly showed that the chromosome complement of the partial amphiploid TAF46 consists of an entire wheat genome plus one synthetic genome consisting of a mixture of six S genome chromosomes and eight J (=E) genome chromosomes derived from Th. intermedium (2n = 6x = 42, JJJsJsSS). There were no Js genome chromosomes present in TAF46. The J genome chromosomes present in TAF46 displayed a unique GISH hybridization pattern with the S genomic DNA probe, in which S genome DNA strongly hybridized at the terminal regions and weakly hybridized over the remaining parts of the chromosomes. This provides a diagnostic marker for distinguishing J genome chromosomes from Js or S genome or wheat ABD genome chromosomes. The genomic origin of the alien chromosomes present in the six derived chromosome addition lines were identified by their characteristic GISH hybridization patterns with S genomic DNA probe. GISH analysis showed that addition lines L1, L2, L3, and L5 carried one pair of J genome chromosomes, while addition lines L4 and L7 each carried one pair of S genome chromosomes. GISH patterns detected by the S genome probe on addition line of L1 were identical to those of the J genome chromosomes present in the partial amphiploid TAF46, suggesting that these chromosomes were not structurally altered when they were transferred from TAF46 to addition lines.Key words: GISH, genomic composition, addition lines, Thinopyrum intermedium, partial amphiploid.

Molecular characterization of a wheat – Thinopyrum ponticum partial amphiploid and its derivatives for resistance to leaf rust

Genome ◽  
2003 ◽  
Vol 46 (5) ◽  
pp. 906-913 ◽  
Author(s):  
Hongjie Li ◽  
Qin Chen ◽  
Robert L Conner ◽  
Beihai Guo ◽  
Yanmin Zhang ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Leaf Rust ◽  
Genomic Dna ◽  
Genomic In Situ Hybridization ◽  
Addition Lines ◽  
Thinopyrum Ponticum ◽  
Sequence Tagged Site ◽  
Partial Amphiploid ◽  
Gish Analysis

Leaf rust (caused by Puccinia triticina Eriks.) occurs annually in most wheat-growing areas of the world. Thinopyrum ponticum (Podp.) Z.-W. Liu & R.-C. Wang has provided several leaf rust resistance genes to protect wheat from this fungal disease. Three chromosome substitution lines, Ji806, Ji807, and Ji859, and two chromosome addition lines, Ji791 and Ji924, with a winter growing habit were developed from crosses between wheat (Triticum aestivum L. em Thell.) and the wheat – Th. ponticum partial amphiploid line 693. These lines were resistant to leaf rust isolates from China. Sequence-tagged site (STS) analysis with the J09-STS marker, which is linked to the gene Lr24, revealed that the partial amphiploid line 693 and all of the substitution and addition lines carried gene Lr24. Genomic in situ hybridization (GISH) analysis was carried out on chromosome preparations using total genomic DNA from Pseudoroegneria strigosa (M. Bieb) A. Löve (St genome, 2n = 14) as a probe in the presence of total genomic DNA from T. aestivum 'Chinese Spring' wheat (ABD genomes, 2n = 42). The GISH analysis demonstrated that these lines had a pair of chromosomes displaying the typical pattern of a Js genome chromosome. This indicates that the chromosome that carries gene Lr24 belonged to the Js genome of Th. ponticum. In addition to 40 wheat chromosomes, eight Js and eight J genome chromosomes were also differentiated by GISH in the partial amphiploid line 693. Since most sources of Lr24 have a red grain color, the white-colored seeds in all of these substitution and addition lines, together with high protein content in some of the lines, make them very useful as a donor source for winter wheat breeding programs.Key words: Lr24, genomic in situ hybridization, sequence-tagged site, random amplified polymorphic DNA.

Genomic in situ hybridization to sectioned nuclei shows chromosome domains in grass hybrids

1990 ◽  
Vol 95 (3) ◽  
pp. 335-341
Author(s):  
A.R. Leitch ◽  
W. Mosgoller ◽  
T. Schwarzacher ◽  
M.D. Bennett ◽  
J.S. Heslop-Harrison
Keyword(s):  
In Situ Hybridization ◽  
Genomic Dna ◽  
Microscope Level ◽  
Hordeum Chilense ◽  
Root Tip ◽  
Interphase Nuclei ◽  
Light Microscope Level ◽  
Thin Sections ◽  
Electron Microscopes

In situ hybridization using biotinylated total genomic DNA and avidin detection systems was adapted for examination of thin-sectioned plant material in the light and electron microscopes. Root tip material was preserved prior to sectioning, so that the in vivo disposition of the chromatin was maintained. Use of total genomic DNA from Secale africanum as a probe enabled the chromatin from the two parental genomes in the grass hybrid Hordeum chilense × S. africanum to be distinguished. The biotinylated probe preferentially labelled the chromosomes of S. africanum origin. DNA-DNA hybrids were visualized at the light-microscope level by Texas Red fluorescence and at the electron-microscope level by the enzymic precipitation of DAB (diaminobenzidine) or by colloidal gold particles. The use of thin sections allowed the location of probe hybridization to be established unequivocally in both metaphase and interphase nuclei. Analysis of interphase nuclei showed that chromatin originating from the two parental genomes did not intermix but occupied distinct domains.

Genome discrimination by in situ hybridization in Icelandic species of Elymus and Elytrigia (Poaceae: Triticeae)

Genome ◽  
2001 ◽  
Vol 44 (2) ◽  
pp. 275-283 ◽  
Author(s):  
Marian Ørgaard ◽  
Kesara Anamthawat-Jónsson
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Genomic Dna ◽  
Genomic In Situ Hybridization ◽  
Pseudoroegneria Spicata ◽  
Genome Constitution ◽  
Elytrigia Repens ◽  
Elymus Caninus ◽  
Elymus Species

The genome constitution of Icelandic Elymus caninus, E. alaskanus, and Elytrigia repens was examined by fluorescence in situ hybridization using genomic DNA and selected cloned sequences as probes. Genomic in situ hybridization (GISH) of Hordeum brachyantherum ssp. californicum (diploid, H genome) probe confirmed the presence of an H genome in the two tetraploid Elymus species and identified its presence in the hexaploid Elytrigia repens. The H chromosomes were painted uniformly except for some chromosomes of Elytrigia repens which showed extended unlabelled pericentromeric and subterminal regions. A mixture of genomic DNA from H. marinum ssp. marinum (diploid,Xa genome) and H. murinum ssp. leporinum (tetraploid,Xu genome) did not hybridize to chromosomes of the Elymus species or Elytrigia repens, confirming that these genomes were different from the H genome. The St genomic probe from Pseudoroegneria spicata (diploid) did not discriminate between the genomes of the Elymus species, whereas it produced dispersed and spotty hybridization signals most likely on the two St genomes of Elytrigia repens. Chromosomes of the two genera Elymus and Elytrigia showed different patterns of hybridization with clones pTa71 and pAes41, while clones pTa1 and pSc119.2 hybridized only to Elytrigia chromosomes. Based on FISH with these genomic and cloned probes, the two Elymus species are genomically similar, but they are evidently different from Elytrigia repens. Therefore the genomes of Icelandic Elymus caninus and E. alaskanus remain as StH, whereas the genomes of Elytrigia repens are proposed as XXH.Key words: Elymus, Elytrigia, H genome, St genome, in situ hybridization.

The genomic identification of some monosomics of Avena sativa L. cv. Sun II using genomic in situ hybridization

Genome ◽  
1995 ◽  
Vol 38 (4) ◽  
pp. 747-751 ◽  
Author(s):  
J. M. Leggett ◽  
G. S. Markhand
Keyword(s):  
In Situ Hybridization ◽  
Avena Sativa ◽  
Genomic Dna ◽  
Diploid Species ◽  
Genomic In Situ Hybridization ◽  
Chromosome Translocations ◽  
C Genome

Genomic in situ hybridization using total genomic DNA extracted from the C genome diploid species Avena eriantha (2n = 2x = 14, genome CpCp) was used to identify monosomics (2n = 6x − 1 = 41) of the constituent genomes of the hexaploid cultivated oat A. sativa L. cv. Sun II (2n = 6x = 42, genomes AACCDD). The results demonstrate 3 AD/C and 6 C/AD chromosome translocations, indicate that five of the missing monosomics are derived from the C genome, and show that there are duplicates within the partial monosomic series. Chromosome polymorphisms between some monosomic lines are also demonstrated.Key words: Avena, monosomics, genomic in situ hybridization, genomic identification.

Detection of maize DNA sequences amplified in wheat

Genome ◽  
1995 ◽  
Vol 38 (5) ◽  
pp. 946-950 ◽  
Author(s):  
Juan Zhang ◽  
Bernd Friebe ◽  
Bikram S. Gill
Keyword(s):  
Triticum Aestivum ◽  
Zea Mays ◽  
In Situ Hybridization ◽  
Dna Sequences ◽  
Hexaploid Wheat ◽  
Chinese Spring ◽  
Genomic Dna ◽  
Genomic In Situ Hybridization ◽  
Metaphase Chromosomes

Genomic in situ hybridization to somatic metaphase chromosomes of hexaploid wheat cv. Chinese Spring using biotinylated maize genomic DNA as a probe revealed the existence of amplified maize DNA sequences in five pairs of chromosomes. The in situ hybridization sites were located on chromosomes 1A, 7A, 2B, 3B, and 7B. One pair of in situ hybridization sites was also observed in hexaploid oat. The locations and sizes of in situ hybridization sites varied among progenitor species.Key words: Triticum aestivum, Zea mays, shared DNA sequences, genomic in situ hybridization.

Identification of the parental chromosomes of the wheat–alien amphiploid Agrotana by genomic in situ hybridization

Genome ◽  
1995 ◽  
Vol 38 (6) ◽  
pp. 1163-1169 ◽  
Author(s):  
Qin Chen ◽  
R. L. Conner ◽  
A. Laroche
Keyword(s):  
In Situ Hybridization ◽  
Genomic Dna ◽  
Molecular Cytogenetics ◽  
Unknown Origin ◽  
Genomic In Situ Hybridization ◽  
Haynaldia Villosa ◽  
Genomic Relationships ◽  
High Degree ◽  
Donor Species

Labelled total genomic DNA from four alien species, Thinopyrum ponticum (Host) Beauv. (2n = 70, genomes J1J1J1J2J2), Th. bessarabicum (Savul. &Rayss) Love (2n = 14, genome J), Th. elongatum (Host) Beauv. (2n = 14, genome E), and Haynaldia villosa (L.) Schur. (2n = 14, genome V), were used as probes in combination with blocking wheat DNA for in situ hybridization of the chromosomes of Agrotana, a wheat–alien hybrid (2n = 56) of unknown origin. The results showed that genomic DNA probes from Th. ponticum and Th. bessarabicum both clearly revealed 16 alien and 40 wheat chromosomes in Agrotana, indicating that the J genome present in these two species has a high degree of homology with the alien chromosomes in Agrotana. Biotinylated genomic DNA probe from Th. elongatum identified 10 chromosomes from Agrotana, while some regions of six other chromosomes yielded a weak or no signal. The probe from H. villosa produced no differential labelling of the chromosomes of Agrotana. The genomic formula of Agrotana was designated as AABBDDJJ. We suggest that the alien parent donor species of Agrotana is Th. ponticum rather than Th. bessarabicum. Genomic relationships of the three Thinopyrum species are discussed in relation to the distribution of GISH signals in the chromosomes of Agrotana.Key words: Thinopyrum species, wheat–alien amphiploid, genomic DNA probing, in situ hybridization, molecular cytogenetics.

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