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

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.

Genome ◽  
1999 ◽  
Vol 42 (5) ◽  
pp. 821-828 ◽  
Author(s):  
S J Schulte ◽  
S D Rider, Jr. ◽  
J H Hatchett ◽  
J J Stuart

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.


Genome ◽  
2002 ◽  
Vol 45 (5) ◽  
pp. 812-822 ◽  
Author(s):  
Stanley Dean Rider, Jr. ◽  
Weilin Sun ◽  
Roger H Ratcliffe ◽  
Jeffrey J Stuart

AFLP markers in linkage disequilibrium with vH13, an avirulence gene in the Hessian fly (Mayetiola destructor) that conditions avirulence to resistance gene H13 in wheat (Triticum spp.), were discovered by bulked segregant analysis. Five AFLPs were converted into codominant site-specific markers that genetically mapped within 13 cM of this gene. Flanking markers used as probes positioned vH13 near the telomere of the short arm of Hessian fly chromosome X2. These results suggest that the X-linked avirulence genes vH6, vH9, and vH13 are present on Hessian fly chromosome X2 rather than on chromosome X1 as reported previously. Genetic complementation demonstrated that recessive alleles of vH13 were responsible for the H13-virulence observed in populations derived from four different states in the U.S.A.: Georgia, Maryland, Virginia, and Washington. Results support the hypothesis that a gene-for-gene interaction exists between wheat and Hessian fly.Key words: bulked segregant analysis, gene-for-gene interaction, wheat, Triticum, Mayetiola destructor.


Genome ◽  
1994 ◽  
Vol 37 (6) ◽  
pp. 1018-1021 ◽  
Author(s):  
M. Nenno ◽  
K. Schumann ◽  
W. Nagl

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.


Genome ◽  
1999 ◽  
Vol 42 (4) ◽  
pp. 744-751 ◽  
Author(s):  
Anna Zambetaki ◽  
Antigone Zacharopoulou ◽  
Zacharias G. Scouras ◽  
Penelope Mavragani-Tsipidou

Genome ◽  
1998 ◽  
Vol 41 (5) ◽  
pp. 702-708 ◽  
Author(s):  
J.J. Stuart ◽  
S.J. Schulte ◽  
P.S. Hall ◽  
K.M. Mayer

1974 ◽  
Vol 62 (1) ◽  
pp. 215-222 ◽  
Author(s):  
A. G. Gambarini ◽  
F. J. S. Lara

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.


1977 ◽  
Vol 115 (3) ◽  
pp. 539-563 ◽  
Author(s):  
Paul Szabo ◽  
Robert Elder ◽  
Dale M. Steffensen ◽  
Olke C. Uhlenbeck

Chromosoma ◽  
1992 ◽  
Vol 101 (7) ◽  
pp. 448-455 ◽  
Author(s):  
A. Zacharopoulou ◽  
M. Frisardi ◽  
C. Savakis ◽  
A. S. Robinson ◽  
P. Tolias ◽  
...  

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