PAG-I Plant Genome I Conference

Town & Country Conference Center, San Diego, CA, November, 1992.


PG-I: IDENTIFICATION OF RAPD POLYMORPHISMS AMONG INTROGRESSED FLAX LINES CONTAINING THE M RUST RESISTANCE GENES, AND THE CLONING OF THE PCR PRODUCTS

IDENTIFICATION OF RAPD POLYMORPHISMS AMONG INTROGRESSED FLAX LINES CONTAINING THE M RUST RESISTANCE GENES, AND THE CLONING OF THE PCR PRODUCTS.

Chris Cullis and Didahally Govindaraju, Biology Department, Case Western Reserve University, Cleveland, OH 44106.


Most of the flax rust resistance genes have been incorporated into a Bison background by Flor. A series of 7 lines, each containing a single M gene, was developed by backcrossing to Bison 8 to 14 times. These lines and the source parents along with Bison were used in a comparison to identify RAPD polymorphisms based on screening with 100 random 10-mers as primers. A number of reproducible polymorphisms were identified between the introgressed lines and Bison. The polymorphic pattern was checked against the relevant source parent to ensure that the difference was consistent with it being derived from the source parent. The most polymorphisms were observed with the line which had only been backcrossed to Bison 8 times compared to those backcrossed 12 or 14 times. The polymorphic bands from Bison and/or the differential were excised and cloned using the TA cloning system from Invitrogen. The cloned amplified fragments will be used to test whether or not they highlight useful RFLPS. In addition, random PCR amplification products have been cloned to determine the relative frequency of repetitive and low copy sequences being amplified in flax using random 10-mers as primers.


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