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Detection of DNA sequence polymorphisms among wheat varieties
S He1, H Ohm, S Mackenzie
1Department of Agronomy, Purdue University, 47907, West Lafayette, IN, USA.
Summary
A new DNA marker strategy combines randomly amplified polymorphic DNA (RAPD) and denaturing gradient gel electrophoresis (DGGE) for efficient wheat genetic analysis. This method reveals high levels of polymorphism, aiding in wheat breeding and genetic mapping.
Area of Science:
- Plant Genetics
- Molecular Biology
- Biotechnology
Background:
- Common wheat (Triticum aestivum) exhibits low levels of restriction fragment length polymorphism, hindering genetic analysis in self-pollinated crops.
- Developing efficient DNA marker strategies is crucial for understanding genetic diversity and improving crop traits.
Purpose of the Study:
- To develop a rapid and efficient DNA marker detection strategy for high-resolution polymorphism analysis in wheat.
- To overcome the limitations of existing methods for detecting genetic variation in closely related wheat lines.
Main Methods:
- Combined randomly amplified polymorphic DNA (RAPD) with denaturing gradient gel electrophoresis (DGGE).
- Utilized polymerase chain reaction (PCR) amplifications with 65 primer combinations.
- Applied the technique to differentiate wheat genotypes and analyze a recombinant inbred population.
Main Results:
- Over 38% of primer combinations produced detectable and reproducible DNA polymorphisms between wheat lines.
- A high level of polymorphism was observed among commercial wheat varieties and breeding lines.
- Demonstrated feasibility for genome mapping and detecting polymorphisms in a recombinant inbred population.
Conclusions:
- The developed RAPD-DGGE system offers a robust method for DNA polymorphism detection in wheat.
- This strategy facilitates pedigree analysis, fingerprinting, and high-density genetic map construction.
- The approach simplifies marker-based selection in plant breeding programs by avoiding radioactive isotopes and complex DNA extraction.
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