AFLP and STS tagging of Lr19, a gene conferring resistance to leaf rust in wheat |
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| Authors: | R Prins J Z Groenewald G F Marais J W Snape R M D Koebner |
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| Institution: | (1) ARC-Small Grain Institute, P.O. Box 3507, Matieland, South Africa, 7602 e-mail: rprins@maties.sun.ac.za Fax: +27-21-8874171, ZA;(2) Department of Genetics, Private Bag X1, Matieland, South Africa, 7602, ZA;(3) Cereals Research Department, John Innes Centre, Norwich Research Park, Colney Lane, Norwich NR4 7UH, UK, GB |
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| Abstract: | Amplified fragment length polymorphism (AFLP) markers were used to enrich the map of the wheat chromosomal region containing
the Thinopyrum-derived Lr19 leaf rust resistance gene. The region closest to Lr19 was targeted through the use of deletion and recombinant lines of the translocated segment. One of the AFLP bands thus identified
was converted into a sequence-tagged-site (STS) marker. This assay generated a 130-bp PCR fragment in all Lr19-carrying lines tested, except for one deletion mutant, while non-carrier template failed to amplify any product. This sequence
represents the first marker to map on the distal side of Lr19 on chromosome 7el1. The conversion process of AFLP fragments to STS markers was technically difficult, mainly because of the presence of contaminating
fragments. Various approaches were taken to reduce the frequency of false positives and to identify the correct clone. We
were able to formulate a general verification strategy prior to clone sequencing. Various other factors causing problems with
converting AFLP bands to an STS assays are also discussed.
Received: 15 September 2000 / Accepted: 5 January 2001 |
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| Keywords: | AFLP Leaf rust Lr19 STS |
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