Document Detail

Development of a PCR marker for rapid identification of the Bt-10 gene for common bunt resistance in wheat.
MedLine Citation:
PMID:  10791808     Owner:  NLM     Status:  MEDLINE    
In western Canada, the Bt-10 resistance gene in wheat (Triticum aestivum) is effective against all the known races of common bunt caused by Tilletia tritici and T laevis. The genotypes of 199 F2 plants, originated from a cross between BW553 containing Bt-10 and the susceptible spring wheat cultivar 'Neepawa,' were established in greenhouse and field inoculation studies. A ratio of 1:2:1 resistant : heterozygous : susceptible was observed for bunt reaction, indicating that Bt-10 was expressed in a partially dominant fashion. A polymorphic DNA fragment, amplified using RAPD, and previously shown to be linked to Bt-10 was sequenced and SCAR (sequence characterized amplified region) primers devised. However, SCAR primers failed to amplify the polymorphic fragment. Restriction of PCR products with DraI revealed a polymorphic fragment of 490 bp resulting from a single base pair difference between lines possessing Bt-10 and those lacking the gene. As per the base pair difference, FSD and RSA primers were designed to generate a 275-bp polymorphic DNA fragment. Both 275- and 490-bp polymorphic fragments were present in all of the 22 cultivars known to carry Bt-10, and absent in all 16 cultivars lacking Bt-10. A 3:1 ratio was observed for presence: absence of the 275-bp marker in the F2 population. Using Southern analysis, the 490-bp fragment was effective in differentiating homozygous resistant plants from those heterozygous for Bt-10, based on its presence and the hybridization signal strength. A 1:2:1 resistant : heterozygous : susceptible ratio was also observed for the molecular marker and corresponded to 88% of the phenotypes deduced from the original F2 population. The molecular marker was estimated to be between 1.1 cM and 6.5 cM away from the Bt-10 resistance gene, based on the segregation analysis. Segregation analyses of Bt-10 and the 275-bp marker, evaluated in three different Canada Prairie Spring (CPS) wheat populations, demonstrated a segregation ratio of 3:1 for the molecular marker in two of the populations. These results demonstrated that the PCR marker system using the FSD and RSA primer pair permitted a rapid and reliable identification of individual lines carrying the Bt-10 gene for resistance to common bunt.
A Laroche; T Demeke; D A Gaudet; B Puchalski; M Frick; R McKenzie
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Genome / National Research Council Canada = Génome / Conseil national de recherches Canada     Volume:  43     ISSN:  0831-2796     ISO Abbreviation:  Genome     Publication Date:  2000 Apr 
Date Detail:
Created Date:  2000-06-26     Completed Date:  2000-06-26     Revised Date:  2008-11-21    
Medline Journal Info:
Nlm Unique ID:  8704544     Medline TA:  Genome     Country:  CANADA    
Other Details:
Languages:  eng     Pagination:  217-23     Citation Subset:  IM    
Agriculture and Agri-Food Canada, Lethbridge Research Centre, AB.
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MeSH Terms
Blotting, Southern
Cloning, Molecular
Genes, Plant*
Genetic Markers*
Immunity, Innate / genetics
Plant Diseases / genetics*
Polymorphism, Genetic
Polymorphism, Restriction Fragment Length
Random Amplified Polymorphic DNA Technique*
Recombination, Genetic
Triticum / genetics*
Reg. No./Substance:
0/Genetic Markers

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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