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.

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.

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 relationships among diploid and hexaploid species of Andropogon (Poaceae)

Genome ◽  
2004 ◽  
Vol 47 (6) ◽  
pp. 1220-1224 ◽  
Author(s):  
G Norrmann ◽  
L Hanson ◽  
S Renvoize ◽  
I J Leitch
Keyword(s):  
In Situ Hybridization ◽  
Diploid Species ◽  
Genomic In Situ Hybridization ◽  
South American ◽  
The North ◽  
Hexaploid Species ◽  
Genomic Relationships ◽  
Strong Hybridization ◽  
Taxonomic Implications

Andropogon is a pantropical grass genus comprising 100–120 species and found mainly in the grasslands of Africa and the Americas. While the genomic relationships between many Andropogon species have been resolved by studying chromosome behavior in interspecific hybrids, relationships between the North and South American diploids have remained elusive. Further, the genome composition of two hexaploid species (including the important forage grass Andropogon lateralis Nees) has been unclear because of the strong hybridization barriers that exist between species. Consequently, genomic in situ hybridization was applied to shed light on these issues. The results confirmed that (i) both the South American (Andropogon selloanus (Hack.) Hack., Andropogon macrothrix Trin.) and North American (Andropogon gyrans Michx.) diploid species shared a common S genome and (ii) the S genome comprises just one of the three genomes in the hexaploids A. lateralis Nees and Andropogon bicornis L. The evolutionary and taxonomic implications of these findings are discussed.Key words: Andropogon, polyploidy evolution, Poaceae, genomic in situ hybridization, taxonomy.

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).

Synthesis and cytological characterization of trigeneric hybrids of durum wheat with and without Ph1

Genome ◽  
2004 ◽  
Vol 47 (6) ◽  
pp. 1173-1181 ◽  
Author(s):  
Prem P Jauhar ◽  
M Doğramaci ◽  
T S Peterson
Keyword(s):  
In Situ Hybridization ◽  
Chromosome Pairing ◽  
Genetic Recombination ◽  
Genomic In Situ Hybridization ◽  
Homoeologous Pairing ◽  
Alien Gene Transfer ◽  
Chromosome 5B ◽  
Alien Gene ◽  
Trigeneric Hybrids

Wild grasses in the tribe Triticeae, some in the primary or secondary gene pool of wheat, are excellent reservoirs of genes for superior agronomic traits, including resistance to various diseases. Thus, the diploid wheatgrasses Thinopyrum bessarabicum (Savul. and Rayss) Á. Löve (2n = 2x = 14; JJ genome) and Lophopyrum elongatum (Host) Á. Löve (2n = 2x = 14; EE genome) are important sources of genes for disease resistance, e.g., Fusarium head blight resistance that may be transferred to wheat. By crossing fertile amphidiploids (2n = 4x = 28; JJEE) developed from F1 hybrids of the 2 diploid species with appropriate genetic stocks of durum wheat, we synthesized trigeneric hybrids (2n = 4x = 28; ABJE) incorporating both the J and E genomes of the grass species with the durum genomes A and B. Trigeneric hybrids with and without the homoeologous-pairing suppressor gene, Ph1, were produced. In the absence of Ph1, the chances of genetic recombination between chromosomes of the 2 useful grass genomes (JE) and those of the durum genomes (AB) would be enhanced. Meiotic chromosome pairing was studied using both conventional staining and fluorescent genomic in situ hybridization (fl-GISH). As expected, the Ph1-intergeneric hybrids showed low chromosome pairing (23.86% of the complement), whereas the trigenerics with ph1b (49.49%) and those with their chromosome 5B replaced by 5D (49.09%) showed much higher pairing. The absence of Ph1 allowed pairing and, hence, genetic recombination between homoeologous chromosomes. Fl-GISH analysis afforded an excellent tool for studying the specificity of chromosome pairing: wheat with grass, wheat with wheat, or grass with grass. In the trigeneric hybrids that lacked chromosome 5B, and hence lacked the Ph1 gene, the wheat–grass pairing was elevated, i.e., 2.6 chiasmata per cell, a welcome feature from the breeding standpoint. Using Langdon 5D(5B) disomic substitution for making trigeneric hybrids should promote homoeologous pairing between durum and grass chromosomes and hence accelerate alien gene transfer into the durum genomes.Key words: alien gene transfer, chiasma (xma) frequency, chromosome pairing, fluorescent genomic in situ hybridization (fl-GISH), homoeologous-pairing regulator, specificity of chromosome pairing, wheatgrass.

Genome constitutions ofRoegneriaalashanica,R.elytrigioides,R.magnicaespesandR.grandis(Poaceae: Triticeae) via genomic in-situ hybridization

2010 ◽  
Vol 28 (2) ◽  
pp. 206-211 ◽  
Author(s):  
Hai-Qing Yu ◽  
Chun Zhang ◽  
Chun-Bang Ding ◽  
Hai-Qin Zhang ◽  
Yong-Hong Zhou
Keyword(s):  
In Situ Hybridization ◽  
Genomic In Situ Hybridization

scholarly journals Application of the FISH Technique to Visualize Sex Chromosomes in Domestic Cat Spermatozoa

Animals ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 2106
Author(s):  
Barbara Kij-Mitka ◽  
Halina Cernohorska ◽  
Svatava Kubickova ◽  
Sylwia Prochowska ◽  
Wojciech Niżański ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Sex Chromosomes ◽  
Chromosomal Abnormalities ◽  
Molecular Cytogenetics ◽  
Molecular Probes ◽  
Domestic Cat ◽  
Fish Technique ◽  
Y Chromosomes

Fluorescence in situ hybridization is a molecular cytogenetics technique that enables the visualization of chromosomes in cells via fluorescently labeled molecular probes specific to selected chromosomes. Despite difficulties in carrying out the FISH technique on sperm, related to the need for proper nuclear chromatin decondensation, this technique has already been used to visualize chromosomes in human, mouse, cattle, swine, horse, and dog spermatozoa. Until now, FISH has not been performed on domestic cat sperm; therefore, the aim of this study was to visualize sex chromosomes in domestic cat sperm. The results showed the presence of X and Y chromosomes in feline spermatozoa. The procedure used for sperm decondensation and fluorescence in situ hybridization was adequate to visualize chromosomes in domestic cat spermatozoa and, in the future, it may be used to determine the degree of chromosomal abnormalities in these gametes.

scholarly journals Comparative molecular cytogenetics in Melipona Illiger species (Hymenoptera, Apidae)

Sociobiology ◽  
2018 ◽  
Vol 65 (4) ◽  
pp. 696 ◽  
Author(s):  
Vanderly Andrade-Souza ◽  
Olivia Maria Pereira Duarte ◽  
Cinthia Caroline Cardoso Martins ◽  
Igor Silva Santos ◽  
Márcio Gilberto Cardoso Costa ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Chromosome Number ◽  
Fluorescent In Situ Hybridization ◽  
Molecular Cytogenetics ◽  
Fluorochrome Staining ◽  
Rdna Sites ◽  
Cytogenetic Studies ◽  
Melipona Species ◽  
Almost All

Cytogenetic studies in Melipona are scarce with only 24 species analyzed cytogenetically. Of these, six species had the rDNA sites physically mapped and characterized by Fluorescent in situ Hybridization (fish). The aim of this study was to perform karyotype analyzes on Melipona species from different regions of Brazil, with a greater sampling representative of the Amazonian fauna and using conventional, fluorochrome staining and FISH with heterologous rDNA probes. The predominant chromosome number was 2n = 18, however, the subspecies M. seminigra abunensis and M. s. pernigra showed 2n = 22 chromosomes. The karyotypes were symmetrical, however M. bicolor, M. quadrifasciata, M. flavolineata, M. fuscopilosa, M. nebulosa presented the first pair heteromorphic in length. CMA3+ blocks also exhibited heteromorphism of size and in almost all cases coincided with rDNA sites, except for M. crinita and M. nebulosa, which presented additional non-coincident CMA3+ blocks. The CMA/ rDNA sites were terminal and interstitial in species with high heterochromatic content, and pericentromeric in those species with low heterochromatic content. In addition to pointing out cytogenetic features of cytotaxonomic importance, the reorganization of the genome in Melipona is discussed.

scholarly journals Detection of trisomy 12 by fluorescent in situ hybridization (FISH) in chronic lymphocytic leukemia

2000 ◽  
Vol 23 (3) ◽  
pp. 531-533 ◽  
Author(s):  
Maria de Lourdes L.F. Chauffaille ◽  
Eliana Azevedo Marques ◽  
Jose Salvador Rodrigues de Oliveira ◽  
Maria Madalena Rodrigues ◽  
Maria Stella Figueiredo ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Chronic Lymphocytic Leukemia ◽  
Fluorescent In Situ Hybridization ◽  
Molecular Cytogenetics ◽  
Lymphocytic Leukemia ◽  
Specific Method ◽  
Alpha Satellite ◽  
Trisomy 12 ◽  
Mature Lymphocyte

Chronic lymphocytic leukemia (CLL) presents a varying incidence of karyotypic abnormalities whose detection is complicated by difficulties in obtaining mitosis for analysis in this type of mature lymphocyte disorder. Since the introduction of molecular cytogenetics (FISH = fluorescent in situ hybridization), applying centromeric probes for chromosome 12 has made it possible to detect a higher percentage of trisomy 12 cases. The objective of the present study was to detect trisomy 12 by FISH (alpha satellite probe) in 13 patients with CLL whose karyotypes by G-banding were either normal or inadequate. Using this method trisomy 12 was detected in three patients in a percentage of positive cells varying from 55.5% to 79%, showing that FISH is a sensitive and highly specific method for trisomy detection and should be routinely performed when the karyotype is normal.

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