GISHGenomic in situ hybridization reveals cryptic genetic differences between maize and its putative wild progenitor Zea mays subsp. parviglumis

Genome ◽  
2004 ◽  
Vol 47 (5) ◽  
pp. 947-953 ◽  
Author(s):  
G Gonzalez ◽  
V Confalonieri ◽  
C Comas ◽  
C A Naranjo ◽  
L Poggio
Keyword(s):  
Zea Mays ◽  
In Situ Hybridization ◽  
Genomic Dna ◽  
Molecular Cytogenetics ◽  
Blocking Agent ◽  
Genomic In Situ Hybridization ◽  
Evolutionary Relationships ◽  
Wild Progenitor ◽  
Chromosome Regions

The aim of this paper is to test with genomic in situ hybridization the genomic affinities between maize and its putative progenitor Zea mays subsp. parviglumis. Blocking procedures were applied for the purpose of improving discrimination among chromosome regions. Unlabeled genomic DNA from Z. mays subsp. parviglumis as a blocking agent and labeled genomic DNA from maize were hybridized on maize chromosomes. On the other hand, mitotic metaphases from Z. mays subsp. parviglumis were blocked with unlabeled genomic DNA of maize and hybridized with labeled genomic DNA from Z. mays subsp. parviglumis. Both experiments showed that either maize or Z. mays subsp. parviglumis chromosomes have their own unique sequences. This means an unexpected degree of divergence if Z. mays subsp. parviglumis is the only progenitor of maize, a result that is discussed in relation to our previous genomic in situ hybridization observations and to the different scenarios proposed about the origin of maize.Key words: evolutionary relationships, Zea mays subsp. mays, teosinte, Tripsacum, molecular cytogenetics, genomic in situ hybridization (GISH).

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.

Genomic in situ hybridization (GISH) of Tripsacum dactyloides and Zea mays ssp. mays with B chromosomes

Genome ◽  
1999 ◽  
Vol 42 (4) ◽  
pp. 687-691 ◽  
Author(s):  
L. Poggio ◽  
V. Confalonieri ◽  
C. Comas ◽  
A. Cuadrado ◽  
N. Jouve ◽  
...  
Keyword(s):  
Zea Mays ◽  
In Situ Hybridization ◽  
Genomic In Situ Hybridization ◽  
B Chromosomes ◽  
Tripsacum Dactyloides

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.

Molecular cytogenetic analysis of durum wheat × tritordeum hybrids

Genome ◽  
1997 ◽  
Vol 40 (3) ◽  
pp. 362-369 ◽  
Author(s):  
J. Lima-Brito ◽  
H. Guedes-Pinto ◽  
G. E. Harrison ◽  
J. S. Heslop-Harrison
Keyword(s):  
In Situ Hybridization ◽  
Plant Breeding ◽  
Durum Wheat ◽  
Dna Sequences ◽  
Genomic Dna ◽  
Tandem Repeats ◽  
Nuclear Architecture ◽  
Molecular Cytogenetics ◽  
Hordeum Chilense

Southern and in situ hybridization were used to examine the chromosome constitution, genomic relationships, repetitive DNA sequences, and nuclear architecture in durum wheat × tritordeum hybrids (2n = 5x = 35), where tritordeum is the fertile amphiploid (2n = 6x = 42) between Hordeum chilense and durum wheat. Using in situ hybridization, H. chilense total genomic DNA hybridized strongly to the H. chilense chromosomes and weakly to the wheat chromosomes, which showed some strongly labelled bands. pHcKB6, a cloned repetitive sequence isolated from H. chilense, enabled the unequivocal identification of each H. chilense chromosome at metaphase. Analysis of chromosome disposition in prophase nuclei, using the same probes, showed that the chromosomes of H. chilense origin were in individual domains with only limited intermixing with chromosomes of wheat origin. Six major sites of 18S–26S rDNA genes were detected on the chromosomes of the hybrids. Hybridization to Southern transfers of restriction enzyme digests using genomic DNA showed some variants of tandem repeats, perhaps owing to methylation. Both techniques gave complementary information, extending that available from phenotypic, chromosome morphology, or isozyme analysis, and perhaps are useful for following chromosomes or chromosome segments during further crossing of the lines in plant breeding programs.Key words: In situ hybridization, molecular cytogenetics, plant breeding, Hordeum chilense, Southern hybridization, durum wheat, hybrids.

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.

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.

Cytological characterization of potato - Solanum etuberosum somatic hybrids and their backcross progenies by genomic in situ hybridization

Genome ◽  
1999 ◽  
Vol 42 (5) ◽  
pp. 987-992 ◽  
Author(s):  
F Dong ◽  
R G Novy ◽  
J P Helgeson ◽  
J Jiang
Keyword(s):  
In Situ Hybridization ◽  
Dna Marker ◽  
Somatic Hybrids ◽  
Molecular Cytogenetics ◽  
Genomic In Situ Hybridization ◽  
Preferential Pairing ◽  
Potato Germplasm ◽  
Solanum Etuberosum

Four somatic hybrids derived from a diploid wild species Solanum etuberosum and a diploid tuber-bearing Solanum clone 463-4, together with five BC1 and three BC2 plants, were analyzed by genomic in situ hybridization (GISH). None of the four somatic hybrids had the expected chromosome constitutions, i.e., 24 chromosomes from each fusion parent. Either one chromosome from S. etuberosum or one from the potato parent 463-4 was lost in the hybrids. Three BC1 plants had exactly one set of S. etuberosum chromosomes. The other two BC1 plants either had one extra or one fewer S. etuberosum chromosome, possibly because their somatic hybrid parents had an extra or had lost one S. etuberosum chromosome. The presence of one set, or close to one set, of S. etuberosum chromosomes in all BC1 plants suggests a preferential pairing and segregation of the S. etuberosum chromosomes in the somatic hybrids. Two of the three BC2 plants had 52 chromosomes, deviating significantly from the expected chromosome number of 48. These results suggest poor pairing between S. etuberosum and S. tuberosum chromosomes in the BC1 plants. The present study demonstrates the importance of combining GISH and DNA marker analysis for a thorough characterization of potato germplasm containing chromosomes from different species.Key words: potato germplasm, Solanum etuberosum, molecular cytogenetics.

Chromosomal characterization in native populations of Elymus scabrifolius from Argentina through classical and molecular cytogenetics (FISH–GISH)

Genome ◽  
2012 ◽  
Vol 55 (8) ◽  
pp. 591-598 ◽  
Author(s):  
P.A. Tomas ◽  
G.E. González ◽  
G.E. Schrauf ◽  
L. Poggio
Keyword(s):  
In Situ Hybridization ◽  
Chromosome Pair ◽  
Molecular Cytogenetics ◽  
Chromosome Morphology ◽  
Genomic In Situ Hybridization ◽  
Dapi Staining ◽  
Accurate Identification ◽  
Native Populations ◽  
Strong Hybridization

The karyotype of Elymus scabrifolius (Döll) J.H. Hunz. (2n = 4x = 28) was investigated by DAPI staining and in situ hybridization. All the accessions studied presented a symmetric and uniform karyotype constituted by 9m+2m–sm+3sm. DAPI stain showed 1–7 conspicuous bands in all the chromosomes and polymorphisms between accessions. FISH experiments carried out with 45S rDNA as probe (pTa71) showed strong hybridization signals on the metacentric SAT-chromosome pair 8; the submetacentric SAT-chromosome pair 13 presented weaker hybridization. FISH using pSc119.2 clone as probe identified five chromosome pairs. Then, the combination of chromosome morphology, DAPI-staining, and FISH enabled the accurate identification of each chromosome pair in E. scabrifolius. Genomic in situ hybridization (GISH) experiments using Hordeum DNA as probe on mitotic metaphases confirmed unequivocally the presence of the H genome in E. scabrifolius, allowing us to observe six uniformly labeled chromosome pairs and two chromosome pairs with only one arm labeled. The remaining six chromosome pairs were weakly labeled. The rehybridization of FISH slides with Hordeum DNA as probe allow us to assign the genomic provenance of most of the chromosomes in the studied accessions. Moreover, intergenomic rearrangement was detected between genome H and the still unknown progenitor genome.

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