Submitted to: ASA-CSSA-SSSA Annual Meeting Abstracts
Publication Type: Abstract Only
Publication Acceptance Date: 10/31/2004
Publication Date: 10/31/2004
Citation: Park, Y.H., Ulloa, M. 2004. PCR-mediated recombination generated from allotetraploid cotton dna using microsatellite markers.. ASA-CSSA-SSSA Annual Meeting Abstracts.
Technical Abstract: PCR-mediated recombination is known to occur between similar or related DNA template sequences in a single PCR amplification. With 91 high stringent selected microsatellites, recombination between two size-polymorphic homoeologue DNA sequences was tested. We amplified genomic DNAs from two allotetraploid cotton species (G. hirsutum and G. barbadense) and their progenitor diploid species (G. herbaceum and G. ramondii), and a mixture of genomic DNAs from the diploid species. Twenty-seven (29.6%) microsatellite primer sets amplified secondary PCR fragments from the DNA mixture of the two diploid species, in addition to the diploid genome-specific fragments. Eighteen (19.7%) primer sets amplified DNA fragments from allotetraploid species that were similar in size to the secondary PCR fragments observed from the diploid DNA mixture. Cloning of the secondary PCR fragments and sequence comparisons to subgenome specific fragments is being conducted. Current study will 1) elucidate the sequence structure of the secondary PCR fragments and suggest a model for their generation mechanism, 2) examine a potential pitfall of including the PCR recombinants into data analysis in genetic mapping, and 3) test several modified PCR conditions that would minimize the amplification of the recombinants.