The use of random amplified polymorphic DNA markers in wheat

Area of Science:

• * Molecular Biology
• * Genetics
• * Plant Science

Background:

• * Random amplified polymorphic DNA (RAPD) is a molecular marker technique.
• * Its application in wheat genetic analysis requires rigorous optimization.
• * Understanding RAPD behavior is crucial for its effective utilization.

Purpose of the Study:

• * To evaluate the utility of RAPD as a genetic marker system in wheat.
• * To identify factors influencing RAPD pattern reproducibility.
• * To assess the suitability of RAPD for linkage mapping and probe development.

Main Methods:

• * Genomic DNA extraction from wheat varieties and recombinant lines.
• * Optimization of polymerase chain reaction (PCR) conditions, including primer selection, DNA concentration, Mg(2+) concentration, polymerase concentration, and denaturing temperature.
• * Analysis of RAPD amplification products.

Main Results:

• * Reproducible RAPD amplification products were achieved under optimized conditions.
• * Key factors influencing RAPD patterns identified: DNA concentration, Mg(2+) concentration, polymerase concentration, and denaturing temperature.
• * RAPD markers exhibit non-homoeologous, non-dose-responsive, and dominant behavior in wheat.
• * High probability of amplified fragments originating from repetitive DNA.

Conclusions:

• * RAPD markers are less suitable for constructing wheat linkage maps due to their dominant nature.
• * Their use as a source for conventional RFLP probes is limited by repetitive DNA origins.
• * RAPD markers are highly valuable for analyzing genotypes involving manipulation of single chromosomes or chromosome segments.