Identification of wheat and tritordeum chromosomes by genomic in situ hybridization using total Hordeum chilense DNA as probe

Genome ◽  
1999 ◽  
Vol 42 (6) ◽  
pp. 1194-1200 ◽  
Author(s):  
M J González ◽  
A Cabrera
Keyword(s):  
Triticum Aestivum ◽  
In Situ Hybridization ◽  
Hexaploid Wheat ◽  
Chromosome Complement ◽  
Dna Probe ◽  
Hordeum Chilense ◽  
B Genome ◽  
A Genome ◽  
The Individual

Total genomic Hordeum chilense DNA probe was hybridized to somatic chromosome spreads of Triticum aestivum 'Chinese Spring' and to four advanced tritordeum lines, the latter being the fertile amphiploid between H. chilense and durum wheat (2n = 6x = 42, AABBHchHch). The probe hybridized strongly to the B-genome chromosomes and to one or two bands on the A-genome chromosomes present in both wheat and tritordeum alloploids. Bands on chromosomes 1D, 2D, and 7D from hexaploid wheat were also detected. Genomic H. chilense DNA probe identified 16 chromosome pairs of the chromosome complement of hexaploid wheat and all A- and B-genome chromosomes present in the tritordeum amphiploids. The in situ hybridization patterns observed correspond to those previously reported in wheat by both N-banding and in situ hybridization with the GAA-satellite sequence (Pedersen and Langridge 1997), allowing the identification of these chromosomes. Variation among the tritordeum amphiploids for hybridization sites on chromosomes 2A, 4A, 6A, 7A, 4B, 5B, and 7B was observed. Despite of this polymorphism, all lines shared the general banding pattern. When used as probe, total H. chilense genomic DNA labeled the H. chilense chromosomes over their lengths allowing the identification of 14 H. chilense chromosomes present in the tritordeum amphiploids. In addition, chromosome-specific telomeric, interstial, and centromeric hybridization sites were observed. These hybridization sites coincide with N-banded regions in H. chilense allowing the identification of the individual H. chilense chromosomes in one of the amphiploid. The N-banded karyotypes of H. chilense (accessions H1 and H7) are presented.Key words: Hordeum chilense, Triticum aestivum, chromosome identification, in situ hybridization, N-banding.

scholarly journals The effect of avidin-biotin interactions in detection systems for in situ hybridization.

1992 ◽  
Vol 40 (1) ◽  
pp. 135-141 ◽  
Author(s):  
E J Speel ◽  
B Schutte ◽  
F C Ramaekers ◽  
A H Hopman
Keyword(s):  
In Situ Hybridization ◽  
Network Formation ◽  
Transitional Cell ◽  
Fluorescein Isothiocyanate ◽  
Staining Technique ◽  
Dna Probe ◽  
Detection Systems ◽  
Amplification Method ◽  
The Individual

The effect of avidin-biotin interactions in several detection systems for the non-radioactive in situ hybridization (ISH) technique was studied in a model system using a transitional cell carcinoma line and a biotinylated DNA probe. We performed fluorescence ISH to unravel the individual steps in a sensitive and frequently used amplification method which makes use of the alternating cytochemical detection layers of fluorescein isothiocyanate-conjugated avidin (AvFITC) and biotinylated goat anti-avidin (BioGAA) antibodies to detect the hybridized and biotinylated probe. Our experiments revealed that BioGAA antibodies bind with their antigen binding sites and not with their biotin moieties to avidin molecules that have already interacted with the DNA probe. The probable working mechanism of this amplification method is presented in a model. Furthermore, we used a peroxidase staining technique to compare with each other the sensitivity of several other detection systems in which avidin-biotin interactions play an important role, e.g., the avidin-biotinylated peroxidase complex (ABC) system. The experiments show that avidin molecules can not be efficiently used to interconnect two biotinylated molecular layers, since their introduction leads to firmly closed cytochemical networks. Such a closed network is already formed between the hybridized and biotinylated DNA probe and a first detection layer of avidin molecules, as appears from the finding that biotinylated molecules could hardly be coupled to these avidin molecules in a following detection layer. Therefore, the results presented here provide us with new insight into the molecular basis of cytochemical network formation. This will enable us to choose the proper procedures for increasing the sensitivity of ISH detection systems.

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 evidence on the origin of wheat chromosomes 4A and 4B

Genome ◽  
1990 ◽  
Vol 33 (1) ◽  
pp. 30-39 ◽  
Author(s):  
J. Dvořák ◽  
P. Resta ◽  
R. S. Kota
Keyword(s):  
In Situ Hybridization ◽  
Repeated Sequence ◽  
Triticum Aestivum L ◽  
Close Relative ◽  
Molecular Evidence ◽  
Substitution Lines ◽  
Genome Chromosome ◽  
B Genome ◽  
A Genome

The genome allocation of the Triticum aestivum L. chromosomes denoted 4A and 4B was based on an erroneous inference. Since neither chromosome pairs with the chromosomes of putative ancestors of wheat, molecular tools were employed to clarify the origin of the two chromosomes. Disomic substitutions for T. aestivum chromosomes 4A or 4B by chromosomes 4 from T. speltoides (Tausch) Gren., a putative ancestor of the wheat B genome, T. longissimum (Schweinf. et Muschl.) Bowden (a close relative of T. speltoides), or T. monococcum L. ssp. aegilopoides (Link) Thell., a close relative of the ancestor of the wheat A genome, were produced. The ability of the substituted chromosome to compensate in the disomic substitution lines, the C-banding patterns of the chromosomes, electrophoretic alleles at the Adh-1 and Lpx-1 loci, and in situ hybridization with an interspersed repeated sequence all were consistent in showing that the chromosome previously denoted as 4A belongs to the B genome and the chromosome previously denoted as 4B is a rearranged chromosome of the A genome. Chromosome 4A is consequently reallocated to the B genome and chromosome 4B to the A genome in T. turgidum L. em. Morris et Sears and T. aestivum. To reflect the fact that the chromosome previously denoted as 4B has only a homoeologous relationship to chromosome 4A of T. urartu (the ancestor of the A genome in polyploid wheats), the chromosome is designated 4Aa.Key words: repeated nucleotide sequence, alcohol dehydrogenase, lipoxygenase, in situ hybridization, chromosome evolution.

scholarly journals Equivalence of the A genome of bread wheat and that of Triticum urartu

1976 ◽  
Vol 27 (1) ◽  
pp. 69-76 ◽  
Author(s):  
Victor Chapman ◽  
T. E. Miller ◽  
Ralph Riley
Keyword(s):  
Triticum Aestivum ◽  
Hexaploid Wheat ◽  
Homoeologous Pairing ◽  
Triticum Urartu ◽  
B Genome ◽  
Chromosome 5B ◽  
A Genome ◽  
Telocentric Chromosomes ◽  
Mother Cells ◽  
Pairing Behaviour

SUMMARYLines of Triticum aestivum Chinese Spring (2n = 6x = 42) which were ditelocentric or doubly ditelocentric, in turn, for the 14 chromosomes of the A and B genomes were pollinated by Triticum urartu (2n = 14). The behaviour of the marked telocentric chromosomes was scored in the 14 distinct hybrids obtained from these pollinations. In 6 of the hybrids in which different A genome chromosomes were marked by telocentrics there were from 50 to 80% of the pollen mother cells in which the telocentrics were paired. In the seven hybrids in which different B genome chromosomes were marked the telocentrics were never paired. It was concluded that the genome of T. urartu matched very closely the A genome of hexaploid wheat and that it did not correspond, as had been proposed by Johnson, to the B genome. The pairing behaviour of the 14 T. aestivum × T. urartu hybrids was compared with earlier results obtained from hybrids between T. aestivum and T. boeoticum. It was proposed that the higher trivalent frequencies seen in the T. boeoticum hybrids could be due to homoeologous pairing and that the genotype of T. boeoticum has the capacity partly to suppress the activity of the Ph locus of chromosome 5B of wheat, as a result of which homoeologous pairing is normally prevented.

Simultaneous painting of three genomes in hexaploid wheat by BAC-FISH

Genome ◽  
2004 ◽  
Vol 47 (5) ◽  
pp. 979-987 ◽  
Author(s):  
Peng Zhang ◽  
Wanlong Li ◽  
Bernd Friebe ◽  
Bikram S Gill
Keyword(s):  
Triticum Aestivum ◽  
In Situ Hybridization ◽  
Transposable Elements ◽  
Fluorescence In Situ Hybridization ◽  
Hexaploid Wheat ◽  
D Genome ◽  
Bac Clones ◽  
Dispersed Repeat ◽  
Bac Fish

Fluorescence in situ hybridization (FISH) is widely used in the physical mapping of genes and chromosome landmarks in plants and animals. Bacterial artificial chromosomes (BACs) contain large inserts, making them amenable for FISH mapping. In our BAC-FISH experiments, we selected 56 restriction fragment length polymorphism (RFLP)-locus-specific BAC clones from the libraries of Triticum monococcum and Aegilops tauschii, which are the A- and D-genome donors of wheat (Triticum aestivum, 2n = 6x = 42), respectively. The BAC clone 676D4 from the T. monococcum library contains a dispersed repeat that preferentially hybridizes to A-genome chromosomes, and two BAC clones, 9I10 and 9M13, from the Ae. tauschii library contain a dispersed repeat that preferentially hybridizes to the D-genome chromosomes. These repeats are useful in simultaneously discriminating the three different genomes in hexaploid wheat, and in identifying intergenomic translocations in wheat or between wheat and alien chromosomes. Sequencing results show that both of these repeats are transposable elements, indicating the importance of transposable elements, especially retrotransposons, in the genome evolution of wheat.Key words: bacterial artificial chromosome (BAC), fluorescence in situ hybridization (FISH), transposable elements (TEs), wheat, Triticum aestivum.

Coevolution of A and B genomes in allotetraploid Triticum dicoccoides

Genome ◽  
2000 ◽  
Vol 43 (6) ◽  
pp. 1021-1026 ◽  
Author(s):  
Alexander Belyayev ◽  
Olga Raskina ◽  
Abraham Korol ◽  
Eviatar Nevo
Keyword(s):  
In Situ Hybridization ◽  
Repetitive Sequences ◽  
Triticum Dicoccoides ◽  
Genomic In Situ Hybridization ◽  
Banding Technique ◽  
Genome Sequences ◽  
Polyploid Evolution ◽  
B Genome ◽  
A Genome

Data is presented on the coevolution of A and B genomes in allotetraploid wheat Triticum dicoccoides (2n = 4x = 28, genome AABB) obtained by genomic in situ hybridization (GISH). Probing chromosomes of T. dicoccoides with DNA from the proposed A/B diploid genome ancestors shows evidence of enriching A-genome with repetitive sequences of B-genome type. Thus, ancestral S-genome sequences have spread throughout the AB polyploid genome to a greater extent than have ancestral A-genome sequences. The substitution of part of the A-genome heterochromatin clusters by satellite DNA of the B genome is detected by using the molecular banding technique. The cause may be interlocus concerted evolution and (or) colonization. We propose that the detected high level of intergenomic invasion in old polyploids might reflect general tendencies in speciation and stabilization of the allopolyploid genome.Key words: Triticum, polyploid, evolution, genomic in situ hybridization, repetitive sequences.

Simultaneous discrimination of the three genomes in hexaploid wheat by multicolor fluorescence in situ hybridization using total genomic and highly repeated DNA probes

Genome ◽  
1993 ◽  
Vol 36 (3) ◽  
pp. 489-494 ◽  
Author(s):  
Yasuhiko Mukai ◽  
Yumiko Nakahara ◽  
Maki Yamamoto
Keyword(s):  
Triticum Aestivum ◽  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Common Wheat ◽  
Chinese Spring ◽  
Genomic Dna ◽  
D Genome ◽  
B Genome ◽  
Multicolor Fluorescence

Common wheat, Triticum aestivum, is an allohexaploid species consisting of three different genomes (A, B, and D). The three genomes were simultaneously discriminated with different colors. Biotinylated total genomic DNA of the diploid A genome progenitor Triticum urartu, digoxigenin-labeled total genomic DNA of the diploid D genome progenitor Aegilops squarrosa, and nonlabeled total genomic DNA of one of the possible B genome progenitors Ae. speltoides were hybridized in situ to metaphase chromosome spreads of Triticum aestivum cv. Chinese Spring. For detection, only two fluorochromes, fluorescein and rhodamine, were used. The A, B, and D genomes were simultaneously detected by their yellow, brown, and orange fluorescence, respectively. The genomic fluorescence in situ hybridization pattern of chromosome 4A of cv. Chinese Spring wheat showed that the distal 32% of the long arm was derived from a B genome chromosome. Furthermore, by using two highly repeated sequence probes, pSc 119.2 and pAsl, and two fluorochromes simultaneously, we were able to identify all B and D genome chromosomes and chromosomes 1A, 4A, and 5A of wheat.Key words: common wheat, in situ hybridization, multicolor fluorescence.

Ultrastructural analysis of the spatial distribution of MRNA

1992 ◽  
Vol 50 (1) ◽  
pp. 552-553
Author(s):  
Gary Bassell ◽  
Robert H. Singer
Keyword(s):  
Electron Microscopy ◽  
Spatial Distribution ◽  
In Situ Hybridization ◽  
High Resolution ◽  
Nucleic Acids ◽  
Colloidal Gold ◽  
Dna Probe ◽  
Nucleotide Analogs ◽  
Gold Particles

We have been investigating the spatial distribution of nucleic acids intracellularly using in situ hybridization. The use of non-isotopic nucleotide analogs incorporated into the DNA probe allows the detection of the probe at its site of hybridization within the cell. This approach therefore is compatible with the high resolution available by electron microscopy. Biotinated or digoxigenated probe can be detected by antibodies conjugated to colloidal gold. Because mRNA serves as a template for the probe fragments, the colloidal gold particles are detected as arrays which allow it to be unequivocally distinguished from background.

Chromosomal organization of a sequence related to LTR-like elements of Ty1-copia retrotransposons in Avena species

Genome ◽  
1999 ◽  
Vol 42 (4) ◽  
pp. 706-713 ◽  
Author(s):  
Concha Linares ◽  
Antonio Serna ◽  
Araceli Fominaya
Keyword(s):  
In Situ Hybridization ◽  
Diploid Species ◽  
D Genome ◽  
Specific Sequence ◽  
Chromosomal Organization ◽  
Intergenomic Translocations ◽  
A Genome ◽  
Slot Blot Analysis ◽  
Copia Retrotransposons

A repetitive sequence, pAs17, was isolated from Avena strigosa (As genome) and characterized. The insert was 646 bp in length and showed 54% AT content. Databank searches revealed its high homology to the long terminal repeat (LTR) sequences of the specific family of Ty1-copia retrotransposons represented by WIS2-1A and Bare. It was also found to be 70% identical to the LTR domain of the WIS2-1A retroelement of wheat and 67% identical to the Bare-1 retroelement of barley. Southern hybridizations of pAs17 to diploid (A or C genomes), tetraploid (AC genomes), and hexaploid (ACD genomes) oat species revealed that it was absent in the C diploid species. Slot-blot analysis suggested that both diploid and tetraploid oat species contained 1.3 × 104 copies, indicating that they are a component of the A-genome chromosomes. The hexaploid species contained 2.4 × 104 copies, indicating that they are a component of both A- and D-genome chromosomes. This was confirmed by fluorescent in situ hybridization analyses using pAs17, two ribosomal sequences, and a C-genome specific sequence as probes. Further, the chromosomes involved in three C-A and three C-D intergenomic translocations in Avena murphyi (AC genomes) and Avena sativa cv. Extra Klock (ACD genomes), respectively, were identified. Based on its physical distribution and Southern hybridization patterns, a parental retrotransposon represented by pAs17 appears to have been active at least once during the evolution of the A genome in species of the Avena genus.Key words: chromosomal organization, in situ hybridization, intergenomic translocations, LTR sequence, oats.

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