Recombinant chromosomes of advanced backcross plants between Allium cepa L. and A. fistulosum L. revealed by in situ hybridization

2000 ◽  
Vol 100 (8) ◽  
pp. 1190-1196 ◽  
Author(s):  
A. Hou ◽  
E. B. Peffley
Keyword(s):  
In Situ Hybridization ◽  
Allium Cepa ◽  
Advanced Backcross ◽  
Allium Cepa L

scholarly journals The Power of Genomic in situ Hybridization (GISH) in Interspecific Breeding of Bulb Onion (Allium cepa L.) Resistant to Downy Mildew (Peronospora destructor [Berk.] Casp.)

Plants ◽  
2019 ◽  
Vol 8 (2) ◽  
pp. 36 ◽  
Author(s):  
Ludmila Khrustaleva ◽  
Majd Mardini ◽  
Natalia Kudryavtseva ◽  
Rada Alizhanova ◽  
Dmitry Romanov ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Downy Mildew ◽  
Allium Cepa ◽  
Genomic In Situ Hybridization ◽  
Lethal Gene ◽  
Peronospora Destructor ◽  
Bulb Onion ◽  
Allium Cepa L ◽  
Interspecific Breeding

We exploited the advantages of genomic in situ hybridization (GISH) to monitor the introgression process at the chromosome level using a simple and robust molecular marker in the interspecific breeding of bulb onion (Allium cepa L.) that is resistant to downy mildew. Downy mildew (Peronospora destructor [Berk.] Casp.) is the most destructive fungal disease for bulb onions. With the application of genomic in situ hybridization (GISH) and previously developed DMR1 marker, homozygous introgression lines that are resistant to downy mildew were successfully produced in a rather short breeding time. Considering that the bulb onion is a biennial plant, it took seven years from the F1 hybrid production to the creation of S2BC2 homozygous lines that are resistant to downy mildew. Using GISH, it was shown that three progeny plants of S2BC2 possessed an A. roylei homozygous fragment in the distal region of the long arm of chromosomes 3 in an A. cepa genetic background. Previously, it was hypothesized that a lethal gene(s) was linked to the downy mildew resistance gene. With the molecular cytogenetic approach, we physically mapped more precisely the lethal gene(s) using the homozygous introgression lines that differed in the size of the A. roylei fragments on chromosome 3.

Random amplified polymorphic DNA analysis, genome size, and genomic in situ hybridization of triploid viviparous onions

Genome ◽  
1999 ◽  
Vol 42 (6) ◽  
pp. 1208-1216 ◽  
Author(s):  
Jasna Puizina ◽  
Branka Javornik ◽  
Borut Bohanec ◽  
Dieter Schweizer ◽  
Jolanta Maluszynska ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Genome Size ◽  
Allium Cepa ◽  
Dna Analysis ◽  
Random Amplified Polymorphic Dna ◽  
Genomic In Situ Hybridization ◽  
Allium Species ◽  
Allium Cepa L ◽  
Genomic Dnas

Triploid viviparous onions (Allium cepa L. var. viviparum Metzg. (ALEF.), auct.), (2n = 3x = 24), are known in some countries only as a rare relic crop, while in other parts of the world they are still traditionally or even commercially cultivated. Results indicating an identical random amplified polymorphic DNA (RAPD) banding pattern and the same DNA content (2C = 43.4 pg) establish the high genetic similarity and the unique origin of the Croatian clone Ljutika and the Indian clone Pran. In order to determine the parental Allium species of these natural triploid hybrids, genomic fluorescent in situ hybridization (GISH) was applied. Biotinylated genomic DNAs from six diploid Allium species (A. cepa L., A. fistulosum L., A. roylei Stearn, A. vavilovii M. Pop. et Vved., A. galanthum Kar. et Kir., A. oschaninii O. Fedtsch.) were used as probes in this study. While probes obtained from genomic DNA of A. cepa, A. vavilovii, and A. roylei hybridized to somatic chromosomes of Ljutika probes from A. fistulosum, A. galanthum, and A. oschaninii did not. The DNA probes of A. cepa and A. roylei each completely or predominantly labelled one genome (eight chromosomes). A few chromosomes, the markers of the triploid karyotype, were not completely labelled by any probe applied. Our GISH results indicate that triploid viviparous onions might possess a complex triparental genome organization.Key words: triploid viviparous onions, Allium cepa, Allium roylei, genomic in situ hybridization, genome size, random amplified polymorphic DNA (RAPD).

scholarly journals Cytological Evaluations of Advanced Generations of Interspecific Hybrids Between Allium cepa and Allium fistulosum Showing Resistance to Stemphylium vesicarium

Genes ◽  
2019 ◽  
Vol 10 (3) ◽  
pp. 195 ◽  
Author(s):  
Natalia Kudryavtseva ◽  
Michael J. Havey ◽  
Lowell Black ◽  
Peter Hanson ◽  
Pavel Sokolov ◽  
...  
Keyword(s):  
Allium Cepa ◽  
Interspecific Hybrids ◽  
Molecular Classification ◽  
Allium Fistulosum ◽  
Bulb Onion ◽  
Allium Cepa L ◽  
Disease Resistances ◽  
Interspecific Crossing

Interspecific crossing is a promising approach for introgression of valuable traits to develop cultivars with improved characteristics. Allium fistulosum L. possesses numerous pest resistances that are lacking in the bulb onion (Allium cepa L.), including resistance to Stemphylium leaf blight (SLB). Advanced generations were produced by selfing and backcrossing to bulb onions of interspecific hybrids between A. cepa and A. fistulosum that showed resistance to SLB. Molecular classification of the cytoplasm established that all generations possessed normal (N) male−fertile cytoplasm of bulb onions. Genomic in situ hybridization (GISH) was used to study the chromosomal composition of the advanced generations and showed that most plants were allotetraploids possessing the complete diploid sets of both parental species. Because artificial doubling of chromosomes of the interspecific hybrids was not used, spontaneous polyploidization likely resulted from restitution gametes or somatic doubling. Recombinant chromosomes between A. cepa and A. fistulosum were identified, revealing that introgression of disease resistances to bulb onion should be possible.

scholarly journals MOLECULAR HYBRIDIZATION STUDIES IN ALLIUM

HortScience ◽  
1995 ◽  
Vol 30 (3) ◽  
pp. 435e-435
Author(s):  
Usha S. Kallemuchikkal ◽  
E.B. Peffley
Keyword(s):  
Triticum Aestivum ◽  
In Situ Hybridization ◽  
New Mexico ◽  
Allium Cepa ◽  
Genomic Dna ◽  
Interspecific Hybrids ◽  
Allium Fistulosum ◽  
Plant Materials ◽  
High Identity

Total genomic DNA was isolated from study plants and was hybridized with 32P-labeled Allium fistulosum `Ishikura' genomic DNA; Southern blots were performed. Plant materials were Allium cepa `New Mexico Yellow Grano', the Allium fistulosum `Heshiko' and `Ishikura', and their F1 interspecific (Allium fistulosum × Allium cepa) hybrids. Sequences with high identity to the labeled DNA hybridized strongly (i.e., A. fistulosum `Ishikura' hybridized most strongly to itself, next with A. fistulosum `Heshiko'). The least hybridization was observed when A. fistulosum `Ishikura' was hybridized with A. cepa `New Mexico Yellow Grano'. Intensity of the signals observed when DNA of the F1 interspecific hybrids was probed with the `Ishikura' DNA was as expected, with the signals intermediate between those of A. fistulosum to A. fistulosum and A. fistulosum to A. cepa. A second study was performed to identify additional cytological markers in Allium. The 5srDNA and NOR genes from Triticum aestivum onto onion chromosomes using in situ hybridization. Evidence of hybridizations are the presence of fluorescing areas on the chromosomes.

Ultrastructural rRNA localization in plant cell nucleoli. RNA/RNA in situ hybridization, autoradiography and cytochemistry

1993 ◽  
Vol 106 (4) ◽  
pp. 1333-1346 ◽  
Author(s):  
A. Olmedilla ◽  
P.S. Testillano ◽  
O. Vicente ◽  
M. Delseny ◽  
M.C. Risueno
Keyword(s):  
In Situ Hybridization ◽  
Pollen Grains ◽  
Rrna Gene ◽  
Allium Cepa L ◽  
Root Meristematic Cells ◽  
Long Time ◽  
High Resolution Autoradiography ◽  
Rna In Situ Hybridization ◽  
Lowicryl K4m

The distribution of ribosomal transcripts in the plant nucleolus has been studied by non-isotopic in situ hybridization in ultrathin Lowicryl K4M sections and by high-resolution autoradiography after labelling with tritiated uridine. In parallel, cytochemical techniques were applied to localize RNA on different plant nucleolar components of Allium cepa L. root meristematic cells and Capsicum annuum L. pollen grains. For RNA/RNA in situ hybridization, several biotinylated single-stranded ribosomal RNA probes were used for mapping different fragments of the 18 S and the 25 S rRNA gene transcribed regions. Ribosomal RNAs (from pre-rRNAs to mature 18 and 25 S RNAs) were found in the nucleolus, in the dense fibrillar (DFC) and granular components (GC). Hybridization signal was found at the periphery of some fibrillar centres (FCs) with probes recognizing both 18 and 25 S rRNA sequences. A quantitative study was performed to analyze the significance of this labelling. Incorporation of tritiated uridine into roots was carried out and, later, after a long time-exposure, autoradiography revealed the presence of newly synthesized RNA mainly in the DFC and at the periphery of the FCs. The presence of RNA in these areas was also confirmed by the cytochemical techniques used in this study. Taken together, these data favour the hypothesis that transcription can begin at the periphery of the FCs, although we cannot exclude the possibility that the DFC plays a role in this process.

scholarly journals CHOMOSOMAL LOCATION OF BIOTIN- AND FLUORESCEIN-LABELED DNA BY IN SITU HYBRIDIZATION IN ALLIUM.

HortScience ◽  
1990 ◽  
Vol 25 (9) ◽  
pp. 1076a-1076
Author(s):  
Agnes RICROCH ◽  
Robert J. BAKER ◽  
Ellen B. PEFFLEY
Keyword(s):  
In Situ Hybridization ◽  
Allium Cepa ◽  
Interspecific Hybrid ◽  
Allium Fistulosum ◽  
Hybridization Technique ◽  
Mitotic Chromosomes ◽  
Rdna Sequences ◽  
Labeled Probe ◽  
Genomic Map

Biotin- and fluorescein-labeled probe has been used to map. specific sunflower rDNA sequences by in situ hybridization on mitotic chromosomes of Allium cepa, Allium fistulosum and interspecific hybrid derivatives, There are three hybridization sites in A. cepa and more than six in an interspecific triploid. This in situ hybridization technique offers new cytogenetic markers useful in the construction of a physical genomic map of Allium and offer a means to document introgression of these genomes.

DNA sequence mapping in interphase and metaphase chromosomes by fluorescence in situ hybridization

1992 ◽  
Vol 50 (1) ◽  
pp. 496-497
Author(s):  
Barbara Trask ◽  
Susan Allen ◽  
Anne Bergmann ◽  
Mari Christensen ◽  
Anne Fertitta ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Dna Sequence ◽  
Dna Sequences ◽  
Dual Band ◽  
Nick Translation ◽  
Metaphase Chromosomes ◽  
Band Pass ◽  
Texas Red ◽  
Fluorescent Spot

Using fluorescence in situ hybridization (FISH), the positions of DNA sequences can be discretely marked with a fluorescent spot. The efficiency of marking DNA sequences of the size cloned in cosmids is 90-95%, and the fluorescent spots produced after FISH are ≈0.3 μm in diameter. Sites of two sequences can be distinguished using two-color FISH. Different reporter molecules, such as biotin or digoxigenin, are incorporated into DNA sequence probes by nick translation. These reporter molecules are labeled after hybridization with different fluorochromes, e.g., FITC and Texas Red. The development of dual band pass filters (Chromatechnology) allows these fluorochromes to be photographed simultaneously without registration shift.

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.

Sign in / Sign up

Export Citation Format

Share Document