Molecular cytogenetic analysis of DNA sequences with flanking telomeric repeats inTriticum aestivumcv. Begra

Genome ◽  
2001 ◽  
Vol 44 (1) ◽  
pp. 133-136
Author(s):  
Marta Dobrzanska ◽  
Elzbieta Kraszewska ◽  
Maria Bucholc ◽  
Glyn Jenkins
Keyword(s):  
Triticum Aestivum ◽  
Dna Sequences ◽  
Genomic Dna ◽  
Repeat Unit ◽  
Chromosome Complement ◽  
Mobile Dna ◽  
Chromosomal Distribution ◽  
Telomeric Sequences ◽  
Dna Fragment

A cloned genomic DNA fragment (pTa241) formerly derived from a DNA fraction obtained from isolated nuclei of embryos of a Polish cultivar of wheat (Triticum aestivum cv. Begra) comprises a tandem repeat of the telomeric array CCCTAAA, and hybridizes in situ exclusively to the telomeres of all chromosome arms of the somatic chromosome complement of wheat. A second cloned fragment (pTa637) derived from the same fraction is 637 bp long, flanked by 28 bp of the same telomeric repeat unit, and hybridizes in situ to the entire lengths of all the chromosomes of the complement. The same pattern of hybridization was observed when the flanking telomeric sequences were removed. A third DNA fragment (pTa1439), derived from unfractionated genomic DNA and flanked with 62 bp of the same telomeric unit, showed the same patterns of distribution. Together with additional evidence from Southern analysis, these observations were interpreted to mean that these sequences are associated with mobile DNA elements and are distributed widely throughout the genome. The chromosomal distribution of the non-telomeric parts of the clones is consistent with the dispersed genomic distribution characteristic of transposons and retroelements.Key words: wheat, Triticum aestivum cv. Begra, mobile elements, telomeric DNA sequence, FISH.

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.

Molecular analyses of a repetitive DNA sequence in wheat (Triticum aestivum L.)

Genome ◽  
2000 ◽  
Vol 43 (3) ◽  
pp. 556-563 ◽  
Author(s):  
P P Ueng ◽  
A Hang ◽  
H Tsang ◽  
J M Vega ◽  
L Wang ◽  
...  
Keyword(s):  
Triticum Aestivum ◽  
In Situ Hybridization ◽  
Long Terminal Repeat ◽  
Fluorescence In Situ Hybridization ◽  
Repetitive Sequence ◽  
Genomic Dna ◽  
Repeat Unit ◽  
Terminal Repeat ◽  
Genomic Clones

A repetitive sequence designated WE35 was isolated from wheat genomic DNA. This sequence consists of a 320-bp repeat unit and represents approximately 0.002% of the total wheat DNA. It is unidirectionally distributed either continuously or discretely in the genome. Ladder-like banding patterns were observed in Southern blots when the wheat genomic DNA was restricted with endonuclease enzymes EcoRI, HincII, NciI, and NdeI, which is characteristic for tandemly organized sequences. Two DNA fragments in p451 were frequently associated with the WE35 repetitive unit in a majority of λ wheat genomic clones. A 475-bp fragment homologous to the 5'-end long terminal repeat (LTR) of cereal retroelements was also found in some λ wheat genomic clones containing the repetitive unit. Physical mapping by fluorescence in situ hybridization (FISH) indicated that one pair of wheat chromosomes could be specifically detected with the WE35 positive probe p551. WE35 can be considered a chromosome-specific repetitive sequence. This repetitive unit could be used as a molecular marker for genetic, phylogenetic, and evolutionary studies in the tribe Triticeae.Key words: repetitive sequence, genomic DNA, Triticum aestivum, fluorescence in situ hybridization, long terminal repeat.

scholarly journals Retention of relict satellite DNA sequences in Anemone (Ranunculaceae)

2013 ◽  
Vol 72 (1) ◽  
pp. 1-133 ◽  
Author(s):  
Višnja Besendorfer ◽  
Jelena Mlinarec
Keyword(s):  
Dna Sequences ◽  
Southern Hybridization ◽  
Repeat Unit ◽  
Similar Species ◽  
Genomic Component ◽  
Library Hypothesis ◽  
Species Specific ◽  
Eukaryotic Organisms ◽  
Fish Signal

Abstract Satellite DNAis a genomic component present in virtually all eukaryotic organisms. The turnover of highly repetitive satellite DNAis an important element in genome organization and evolution in plants. Here we study the presence, physical distribution and abundance of the satellite DNAfamily AhTR1 in Anemone. Twenty-two Anemone accessions were analyzed by PCR to assess the presence of AhTR1, while fluorescence in situ hybridization and Southern hybridization were used to determine the abundance and genomic distribution of AhTR1. The AhTR1 repeat unit was PCR-amplified only in eight phylogenetically related European Anemone taxa of the Anemone section. FISH signal with AhTR1 probe was visible only in A. hortensis and A. pavonina, showing localization of AhTR1 in the regions of interstitial heterochromatin in both species. The absence of a FISH signal in the six other taxa as well as weak signal after Southern hybridization suggest that in these species AhTR1 family appears as relict sequences. Thus, the data presented here support the »library hypothesis« for AhTR1 satellite evolution in Anemone. Similar species-specific satellite DNAprofiles in A. hortensis and A. pavonina support the treatment of A. hortensis and A. pavonina as one species, i.e. A. hortensis s.l.

Identification of the entire chromosome complement of bread wheat by two-colour FISH

Genome ◽  
1997 ◽  
Vol 40 (5) ◽  
pp. 589-593 ◽  
Author(s):  
C. Pedersen ◽  
P. Langridge
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Dna Sequences ◽  
Banding Pattern ◽  
Aegilops Tauschii ◽  
Chromosome Complement ◽  
Chromosome Identification ◽  
Satellite Sequence ◽  
Entire Chromosome

Using the Aegilops tauschii clone pAs1 together with the barley clone pHvG38 for two-colour fluorescence in situ hybridization (FISH) the entire chromosome complement of hexaploid wheat was identified. The combination of the two probes allowed easy discrimination of the three genomes of wheat. The banding pattern obtained with the pHvG38 probe containing the GAA-satellite sequence was identical to the N-banding pattern of wheat. A detailed idiogram was constructed, including 73 GAA bands and 48 pAs1 bands. Identification of the wheat chromosomes by FISH will be particularly useful in connection with the physical mapping of other DNA sequences to chromosomes, or for chromosome identification in general, as an alternative to C-banding.Key words: Triticum aestivum, chromosome identification, fluorescence in situ hybridization, repetitive DNA sequences.

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.

MOLECULAR CLONING OF α-AMYLASE GENES FROM DROSOPHILA MELANOGASTER. I. CLONE ISOLATION BY USE OF A MOUSE PROBE

Genetics ◽  
1985 ◽  
Vol 110 (2) ◽  
pp. 299-312
Author(s):  
Robert M Gemmill ◽  
Jack N Levy ◽  
Winifred W Doane
Keyword(s):  
Drosophila Melanogaster ◽  
Dna Sequences ◽  
Genomic Dna ◽  
Cdna Sequence ◽  
Lambda Phage ◽  
Hybridization Probe ◽  
Class A ◽  
Class B ◽  
Small Set

ABSTRACT A cloned ä-amylase cDNA sequence from the mouse is homologous to a small set of DNA sequences from Drosophila melanogaster under appropriate conditions of hybridization. A number of recombinant lambda phage that carry homologous Drosophila genomic DNA sequences were isolated using the mouse clone as a hybridization probe. Putative amylase clones hybridized in situ to one or the other of two distinct sites in polytene chromosome 2R and were assigned to one of two classes, A and B. Clone λDm32, representing class A, hybridizes within chromosome section 53CD. Clone λDm65 of class B hybridizes within section 54A1-B1. Clone λDm65 is homologous to a 1450- to 1500-nucleotide RNA species, which is sufficiently long to code for α-amylase. No RNA homologous to λDm32 was detected. We suggest that the class B clone, λDm65, contains the functional Amy structural gene(s) and that class A clones contain an amylase pseudogene.

scholarly journals Genomic in situ hybridization in interspecific hybrids of scallops (Bivalvia, Pectinidae) and localization of the satellite DNA Cf303, and the vertebrate telomeric sequences (TTAGGG)n on chromosomes of scallop Chlamys farreri (Jones & Preston, 1904)

2018 ◽  
Vol 12 (1) ◽  
pp. 83-95
Author(s):  
Liping Hu ◽  
Liming Jiang ◽  
Ke Bi ◽  
Huan Liao ◽  
Zujing Yang ◽  
...  
Keyword(s):  
Interspecific Hybrids ◽  
Distribution Patterns ◽  
Mitotic Chromosomes ◽  
Chromosomal Distribution ◽  
Satellite Dnas ◽  
Banding Patterns ◽  
Telomeric Sequences ◽  
Specific Distribution ◽  
Species Specific

Mitotic chromosome preparations of the interspecific hybrids Chlamysfarreri (Jones & Preston, 1904) × Patinopectenyessoensis (Jay, 1857), C.farreri × Argopectenirradinas (Lamarck, 1819) and C.farreri × Mimachlamysnobilis (Reeve, 1852) were used to compare two different scallop genomes in a single slide. Although genomic in situ hybridization (GISH) using genomic DNA from each scallop species as probe painted mitotic chromosomes of the interspecific hybrids, the painting results were not uniform; instead it showed species-specific distribution patterns of fluorescent signals among the chromosomes. The most prominent GISH-bands were mainly located at centromeric or telomeric regions of scallop chromosomes. In order to illustrate the sequence constitution of the GISH-bands, the satellite Cf303 sequences of C.farreri and the vertebrate telomeric (TTAGGG)n sequences were used to map mitotic chromosomes of C.farreri by fluorescence in situ hybridization (FISH). The results indicated that the GISH-banding pattern presented by the chromosomes of C.farreri is mainly due to the distribution of the satellite Cf303 DNA, therefore suggesting that the GISH-banding patterns found in the other three scallops could also be the result of the chromosomal distribution of other species-specific satellite DNAs.

Tandem repeat DNA localizing on the proximal DAPI bands of chromosomes in Larix, Pinaceae

Genome ◽  
2002 ◽  
Vol 45 (4) ◽  
pp. 777-783 ◽  
Author(s):  
Masahiro Hizume ◽  
Fukashi Shibata ◽  
Ayako Matsumoto ◽  
Yukie Maruyama ◽  
Eiji Hayashi ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Repetitive Dna ◽  
Genomic Dna ◽  
Repeat Unit ◽  
Blot Hybridization ◽  
Southern Blot Hybridization ◽  
Repeat Sequence ◽  
Proximal Region

Repetitive DNA was cloned from HindIII-digested genomic DNA of Larix leptolepis. The repetitive DNA was about 170 bp long, had an AT content of 67%, and was organized tandemly in the genome. Using fluorescence in situ hybridization and subsequent DAPI banding, the repetitive DNA was localized in DAPI bands at the proximal region of one arm of chromosomes in L. leptolepis and Larix chinensis. Southern blot hybridization to genomic DNA of seven species and five varieties probed with cloned repetitive DNA showed that the repetitive DNA family was present in a tandem organization in genomes of all Larix taxa examined. In addition to the 170-bp sequence, a 220-bp sequence belonging to the same DNA family was also present in 10 taxa. The 220-bp repeat unit was a partial duplication of the 170-bp repeat unit. The 220-bp repeat unit was more abundant in L. chinensis and Larix potaninii var. macrocarpa than in other taxa. The repetitive DNA composed 2.0–3.4% of the genome in most taxa and 0.3 and 0.5% of the genome in L. chinensis and L. potaninii var. macrocarpa, respectively. The unique distribution of the 220-bp repeat unit in Larix indicates the close relationship of these two species. In the family Pinaceae, the LPD (Larix proximal DAPI band specific repeat sequence family) family sequence is widely distributed, but their amount is very small except in the genus Larix. The abundant LPD family in Larix will occur after its speciation.Key words: AT-rich tandem repetitive DNA, fluorescence in situ hybridization, Larix, proximal DAPI band.

Searching for telomeric sequences in two Allium species

Genome ◽  
2001 ◽  
Vol 44 (4) ◽  
pp. 640-643 ◽  
Author(s):  
N Cuñado ◽  
E Sánchez-Morán ◽  
J Barrios ◽  
J L Santos
Keyword(s):  
In Situ Hybridization ◽  
Dna Sequences ◽  
Fluorescent In Situ Hybridization ◽  
Two Dimensional ◽  
Ltr Retrotransposons ◽  
Allium Species ◽  
Telomeric Sequences ◽  
Rdna Sequences ◽  
Dimensional Surface

Some Alliaceae species have no tandemly repeated TTTAGGG sequences. Instead, at the very end of their chromosomes, there are highly repetitive satellite and (or) rDNA sequences. These sequences apparently replace the canonical plant telomeric sequences in these species. A method of preparing two-dimensional surface spreads of plant synaptonemal complexes (SCs), combined with fluorescent in situ hybridization, has revealed that telomeric chromatin is tightly condensed at the ends of SCs in plants and animals. Using this method, we have tested the organization and location of those sequences postulated to cap the chromosomes in two species of the genus Allium: A. cepa and A. altaicum. We have also extended this study to other putative telomere candidates, such as LTR (long terminal repeat) and non-LTR retrotransposons. None of the DNA sequences analyzed showed the characteristic telomeric organization at pachytene.Key words: fluorescent in situ hybridization, meiosis, repetitive DNA, Allium, synaptonemal complex.

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