Karyotypic analysis of Triticum monococcum using standard repetitive DNA probes and simple sequence repeats

2012 ◽  
Vol 60 (2) ◽  
pp. 87-95 ◽  
Author(s):  
M. Megyeri ◽  
A. Farkas ◽  
M. Varga ◽  
G. Kovács ◽  
M. Molnár-Láng ◽  
...  
Keyword(s):  
Repetitive Dna ◽  
Wheat Breeding ◽  
Dna Probes ◽  
Triticum Monococcum ◽  
Karyotypic Analysis ◽  
Hybridization Pattern ◽  
Polyploid Wheat ◽  
Breeding Programmes ◽  
Simple Sequence

Triticum monococcum represents an important source of useful genes and alleles that it would be desirable to use in wheat breeding programmes. The well-defined landmarks on the Am chromosomes could accelerate the targeted introgression of T. monococcum chromatin into the wheat genome.Fluorescence in situ hybridization (FISH) using the repetitive DNA probes pSc119.2, Afa family and pTa71 showed that the pSc119.2 probe was not suitable for the identification of Am chromosomes. In contrast, the whole set of Am chromosomes (especially chromosomes 1, 4, 5 and 7) could be discriminated based on the hybridization pattern of pTa71 and Afa family. In situ hybridization with microsatellite motifs (GAA, CAG, AAC and AGG) proved that SSRs represent additional landmarks for the identification of Am chromosomes. The most promising SSR probes were the GAA and CAG motifs, which clearly discriminated the 6Am chromosome and, when used in combination with the Afa family and pTa71 probes, allowed the whole set of Am chromosomes to be reliably identified.In conclusion, fluorescence in situ hybridization using the repetitive DNA probes Afa family and pTa71, combined with SSR probes, makes it possible to identify the Am chromosomes of T. monococcum and to discriminate them from Au chromosomes in the polyploid wheat background.

Improvement of the agronomic traits of a wheat–barley centric fusion by introgressing the 3HS.3BL translocation into a modern wheat cultivar

Genome ◽  
2014 ◽  
Vol 57 (11/12) ◽  
pp. 601-607 ◽  
Author(s):  
Edina Türkösi ◽  
András Farkas ◽  
Nikolett Réka Aranyi ◽  
Borbála Hoffmann ◽  
Viola Tóth ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Repetitive Dna ◽  
Robertsonian Translocation ◽  
Field Experiments ◽  
Agronomic Traits ◽  
Wheat Cultivar ◽  
Centric Fusion ◽  
Dna Probes ◽  
Modern Wheat

The 3HS.3BL spontaneous Robertsonian translocation obtained from the progenies of wheat–barley (Chinese Spring × Betzes) hybrids backcrossed with wheat line Mv9kr1 was transferred into the modern Martonvásár wheat cultivar Mv Bodri. The translocation was identified with molecular cytogenetic methods. The inheritance of the translocation was traced using genomic in situ hybridization. Fluorescence in situ hybridization using barley subtelomeric (HvT01) and centromere-specific [(AGGGAG)4] repetitive DNA probes confirmed that the complete barley chromosome arm was involved in the Robertsonian translocation. The wheat-specific repetitive DNA probes identified the presence of the whole wheat genome, except the short arm of the 3B chromosome. Genotypes homozygous for the centric fusion were selected, after which morphological analysis was performed on the plants and the yield components were measured in the field during two consecutive vegetative seasons. The introgression of the 3HS.3BL translocation into the modern wheat cultivar Mv Bodri significantly reduced the plant height due to the incorporation of the dwarfing allele RhtD1b. The presence of the 3HS.3BL translocation in the Mv9kr1 and Mv Bodri wheat background improved tillering and seeds per plant productivity in field experiments carried out in Martonvásár and Keszthely, Hungary.

scholarly journals Rapid fluorescence in situ hybridization with repetitive DNA probes: Quantification by digital image analysis

Cytometry ◽  
1994 ◽  
Vol 17 (1) ◽  
pp. 13-25 ◽  
Author(s):  
Dino Celeda ◽  
Klaus Aldinger ◽  
Frank-Martin Haar ◽  
Michael Hausmann ◽  
Markus Durm ◽  
...  
Keyword(s):  
Image Analysis ◽  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Repetitive Dna ◽  
Digital Image ◽  
Digital Image Analysis ◽  
Dna Probes

Cytogenetic and molecular markers for detecting Aegilops uniaristata chromosomes in a wheat background

Genome ◽  
2014 ◽  
Vol 57 (9) ◽  
pp. 489-497 ◽  
Author(s):  
Wenping Gong ◽  
Guangrong Li ◽  
Jianping Zhou ◽  
Genying Li ◽  
Cheng Liu ◽  
...  
Keyword(s):  
Molecular Markers ◽  
Expressed Sequence Tag ◽  
Triticum Turgidum ◽  
Wheat Breeding ◽  
Addition Lines ◽  
Chromosome Engineering ◽  
Aegilops Uniaristata ◽  
Expressed Sequence ◽  
Simple Sequence

Aegilops uniaristata has many agronomically useful traits that can be used for wheat breeding. So far, a Triticum turgidum – Ae. uniaristata amphiploid and one set of Chinese Spring (CS) – Ae. uniaristata addition lines have been produced. To guide Ae. uniaristata chromatin transformation from these lines into cultivated wheat through chromosome engineering, reliable cytogenetic and molecular markers specific for Ae. uniaristata chromosomes need to be developed. Standard C-banding shows that C-bands mainly exist in the centromeric regions of Ae. uniaristata but rarely at the distal ends. Fluorescence in situ hybridization (FISH) using (GAA)8 as a probe showed that the hybridization signal of chromosomes 1N–7N are different, thus (GAA)8 can be used to identify all Ae. uniaristata chromosomes in wheat background simultaneously. Moreover, a total of 42 molecular markers specific for Ae. uniaristata chromosomes were developed by screening expressed sequence tag – sequence tagged site (EST–STS), expressed sequence tag – simple sequence repeat (EST–SSR), and PCR-based landmark unique gene (PLUG) primers. The markers were subsequently localized using the CS – Ae. uniaristata addition lines and different wheat cultivars as controls. The cytogenetic and molecular markers developed herein will be helpful for screening and identifying wheat – Ae. uniaristata progeny.

A Simplified Combination of DNA Probe Preparation and Fluorescence in situ Hybridization

1992 ◽  
Vol 47 (9-10) ◽  
pp. 739-747 ◽  
Author(s):  
Dino Celeda ◽  
Ulrich Bettag ◽  
Christoph Cremer
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Repetitive Dna ◽  
Human Lymphocytes ◽  
Dna Probes ◽  
Dna Probe ◽  
Nick Translation ◽  
Metaphase Chromosomes ◽  
Human Dna

Fluorescence in situ hybridization (FISH) has found widespread applications in cytogenetics. So far the standard protocols for probe amplification (and simultaneous labeling) by PCR, nick translation and in situ hybridization involve different buffer systems leading to a number of time consuming washing steps even before hybridization. In this manuscript we show a fast technique of a close combination of DNA probe preparation and in situ hybridization (ISH). This method was applied to metaphase chromosomes from human lymphocytes fixed on slides. Two specific repetitive DNA probes, the pUC 1.77 DNA probe and the DYZ 1 repetitive DNA fraction were used, amplified and labeled in different ways. Additional experiments with total genomic male human DNA as the DNA probe suggest that this method may be extended to a large variety of other probes. Moreover the ISH technique described does not require toxic denaturing agents, such as formamide.

Sign in / Sign up

Export Citation Format

Share Document