Molecular genetic mapping of three X-linked avirulence genes, vH6, vH9 andvH13, in the Hessian fly

Genome ◽  
1999 ◽  
Vol 42 (5) ◽  
pp. 821-828 ◽  
Author(s):  
S J Schulte ◽  
S D Rider, Jr. ◽  
J H Hatchett ◽  
J J Stuart
Keyword(s):  
In Situ Hybridization ◽  
Molecular Genetic ◽  
Polytene Chromosomes ◽  
Hessian Fly ◽  
Genetic Distances ◽  
Mayetiola Destructor ◽  
Avirulence Genes ◽  
Hessian Fly Resistance ◽  
Relationship Of

Three X-linked avirulence genes, vH6, vH9, and vH13 in the Hessian fly, Mayetiola destructor, confer avirulence to Hessian fly resistance genes H6, H9, and H13 in wheat. We used a combination of two- and three-point crosses to determine the order of these genes with respect to each other, the white eye mutation and three X-linked molecular markers, G15-1, 020, and 021, developed from genomic lambda clones, λG15-1, λ020, and λ021. The gene order was determined to be vH9-vH6-G15-1-w-vH13-020-021. In situ hybridization of λG15-1, λ020, and λ021, on the polytene chromosomes of the Hessian fly salivary gland established their orientation on Hessian fly chromosome X1. Based on the size of the Hessian fly genome, and the genetic distances between markers, the relationship of physical to genetic distance was estimated at no more than 300 kb/cM along Hessian fly chromosome X1, suggesting that map-based cloning of these avirulence genes will be feasible.Key words: Mayetiola destructor, avirulence genes, genetic map, SSCP, in situ hybridization.

Genetic mapping of Hessian fly avirulence gene vH6 using bulked segregant analysis

Genome ◽  
1998 ◽  
Vol 41 (5) ◽  
pp. 702-708 ◽  
Author(s):  
J J Stuart ◽  
S J Schulte ◽  
P S Hall ◽  
K M Mayer
Keyword(s):  
In Situ Hybridization ◽  
Bulked Segregant Analysis ◽  
Polytene Chromosomes ◽  
Hessian Fly ◽  
Mayetiola Destructor ◽  
Dna Polymorphisms ◽  
Avirulence Gene ◽  
Conformational Polymorphism ◽  
Single Stranded Conformational Polymorphism

The Hessian fly, Mayetiola destructor (Say), an important insect pest of wheat, Triticum aestivum L., has a gene-for-gene relationship with wheat: single genes in the insect condition avirulence to specific resistance genes in wheat. We report the discovery of the first molecular genetic marker that is tightly linked to a Hessian fly avirulence gene. This dominant DNA polymorphism (OPG15-1) was identified using bulked segregant analysis and arbitrary primers in polymerase chain reactions. Bulked segregant analysis was modified to accommodate the anomalous chromosome cycle of the Hessian fly. It was used to identify DNA polymorphisms linked to the gene (vH6) that confers avirulence to the resistance gene H6 in wheat. OPG15-1 was cloned and sequenced, and a pair of site-specific primers were designed that converted it into a codominant single-stranded conformational polymorphism. Both OPG15-1 and vH6 were shown to be X-linked, and the genetic distance between the two loci was 2.5 ± 2.5 cM. In situ hybridization to polytene chromosomes of larval salivary glands indicated that OPG15-1 resides near the centromere of Hessian fly chromosome X1.Key words: Mayetiola destructor, avirulence gene, RAPD-PCR, bulked segregant analysis, single-stranded conformational polymorphism, SSCP, in situ hybridization.

Detection of rRNA and phaseolin genes on polytene chromosomes of Phaseolus coccineus by fluorescence in situ hybridization after pepsin pretreatment

Genome ◽  
1994 ◽  
Vol 37 (6) ◽  
pp. 1018-1021 ◽  
Author(s):  
M. Nenno ◽  
K. Schumann ◽  
W. Nagl
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Ribosomal Rna ◽  
Storage Protein ◽  
Seed Storage Protein ◽  
Polytene Chromosomes ◽  
Seed Storage ◽  
Phaseolus Coccineus ◽  
Rna Genes

This is the first report of fluorescence in situ hybridization (FISH) on plant polytene chromosomes. Different protease pretreatments have been tested to improve fluorescence in situ hybridization FISH on polytene chromosomes of a plant, Phaseolus coccineus, with the aim to enable the detection of low-copy genes. The structural preservation of the chromosomes and the distinctness of the FISH signals were comparatively analysed with a probe for the ribosomal RNA genes after digestion with pepsin and trypsin. The pepsin pretreatment resulted in a general loosening of chromatin with good conservation of chromosome morphology and an increased number and density of signal points. The six nucleolus organizers exhibited significant differences in condensation. The pretreatment with pepsin enabled the detection of the low-copy genes encoding the seed storage protein phaseolin.Key words: plant, Leguminosae, ribosomal RNA genes, seed storage protein genes, protease.

scholarly journals A three-dimensional structural dissection of Drosophila polytene chromosomes.

1995 ◽  
Vol 131 (2) ◽  
pp. 279-295 ◽  
Author(s):  
Y Urata ◽  
S J Parmelee ◽  
D A Agard ◽  
J W Sedat
Keyword(s):  
Rna Polymerase Ii ◽  
Polytene Chromosome ◽  
Polytene Chromosomes ◽  
Three Dimensional ◽  
Spatial Relationship ◽  
Band Shape ◽  
Structural Details ◽  
Novel Method ◽  
Relationship Of

We have analyzed the three-dimensional structural details of Drosophila melanogaster polytene chromosome bands and interbands using three-dimensional light microscopy and a novel method of sample preparation that does not involve flattening or stretching the chromosomes. Bands have been visualized in unfixed chromosomes stained with the DNA specific dye 4,6-Diamidino-2-phenylindole (DAPI). Interbands have been visualized using fixed chromosomes that have been immunostained with an antibody to RNA polymerase II. Additionally, these structures have been analyzed using in situ hybridization with probes from specific genetic loci (Notch and white). Bands are seen to be composed of approximately 36 substructural features that measure 0.2-0.4 micron in diameter. We suggest that these substructural features are in fact longitudinal fibers made up of bundles of chromatids. Band shape can be a reproducible characteristic of a particular band and is dependent on the spatial relationship of these bundles, varying from bands with a uniform distribution of bundles to bands with a peripheral concentration of chromatin. Interbands are composed of bundles of chromatids of a similar size and number as those seen in the bands. The distribution of bundles is similar between a band and the neighboring interband, implying that there is a long range organization to the DNA that includes both the coding and the noncoding portions of genes. Finally, we note that the polytene chromosome has a circular shape when viewed in cross section, whether there are one or two homologs present.

The genome of the olive fruit fly Bactrocera oleae: localization of molecular markers by in situ hybridization to the salivary gland polytene chromosomes

Genome ◽  
1999 ◽  
Vol 42 (4) ◽  
pp. 744-751 ◽  
Author(s):  
Anna Zambetaki ◽  
Antigone Zacharopoulou ◽  
Zacharias G. Scouras ◽  
Penelope Mavragani-Tsipidou
Keyword(s):  
Salivary Gland ◽  
Molecular Markers ◽  
In Situ Hybridization ◽  
Polytene Chromosomes ◽  
Fruit Fly ◽  
Bactrocera Oleae ◽  
Olive Fruit Fly ◽  
Olive Fruit

scholarly journals UNDER-REPLICATION OF RIBOSOMAL CISTRONS IN POLYTENE CHROMOSOMES OF RHYNCHOSCIARA

1974 ◽  
Vol 62 (1) ◽  
pp. 215-222 ◽  
Author(s):  
A. G. Gambarini ◽  
F. J. S. Lara
Keyword(s):  
Salivary Gland ◽  
In Situ Hybridization ◽  
Xenopus Laevis ◽  
Related Species ◽  
Polytene Chromosomes ◽  
Chromosome X ◽  
Heterologous Hybridization ◽  
Ribosomal Cistrons ◽  
Rhynchosciara Americana

DNA preparations obtained from several tissues of Rhynchosciara americana and two related species, R. milleri and R. papaveroi, were hybridized to R. americana rRNA. The percentage of hybridization was found to be higher in tissues with low polyteny than in tissues with high polyteny, suggesting a relationship between the amount of rDNA and the tissue polyteny. This could be explained by under-replication of ribosomal cistrons in polytene cells, such as those from the salivary gland. Only slight tissue-dependent changes in the percentages of hybridization can be observed in heterologous hybridization using Xenopus laevis rRNA. The possibility that these experiments could not detect differences in the amount of ribosomal cistrons among tissues is discussed. The female:male ratio for the percentages of hybridization in the salivary gland of R. americana agrees with the results obtained by in situ hybridization experiments (16, 17) which have shown that the rRNA cistrons are distributed among chromosomes other than chromosome X.

Quantitative in situ hybridization of ribosomal RNA species to polytene chromosomes of Drosophila melanogaster

1977 ◽  
Vol 115 (3) ◽  
pp. 539-563 ◽  
Author(s):  
Paul Szabo ◽  
Robert Elder ◽  
Dale M. Steffensen ◽  
Olke C. Uhlenbeck
Keyword(s):  
Drosophila Melanogaster ◽  
In Situ Hybridization ◽  
Ribosomal Rna ◽  
Polytene Chromosomes
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