PAG-III Plant Genome III Conference

Town & Country Conference Center, San Diego, CA, January, 1995.

PG-III: 101 - DNA MARKERS GENERATED BY RESTRICTION SITE PRIMERS IN WHEAT

DNA MARKERS GENERATED BY RESTRICTION SITE PRIMERS IN WHEAT.

Ismail Dweikat and Herbert Ohm, Department of Agronomy, Purdue University, West Lafayette, IN 47907.

The level of DNA polymorphism in wheat is low among closely related lines. The use of RAPD-DGGE (random amplified polymorphic DNA analysis in combination with denaturing gradient gel electrophoresis) improved the detection of DNA polymorphism significantly over resolution using agarose gels. Currently we are using AFLPs (amplified fragment length polymorphism) resolved by DGGE as an alternate methodology for detecting DNA polymorphisms closely associated with Hessian fly resistance in wheat near isolines. We have tested 9 Pstl and 8 EcoRl primers. With these primers we were able to identify polymorphic fragments associated with resistance genes H6 and H16 (one each). In addition to AFLP markers we have designed 10-mer primers based on the recognition sites for PstI, EcoRl and Sacl. We have tested 9 different primers encompassing these restriction sites in all possible combinations and 4 polymorphisms were identified, each associated with an individual resistance genes, H5, H6, H10, or H16.

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