Development of TaqMan PCR Zygosity Assay to Detect the Maize bm3 Mutant.

 

Brown midrib (BMR) corn varieties have a natural mutation that results in low lignin in their cell walls, leading to increased digestibility, which is a highly desirable trait for silage maize production.  Based on the sequence information from the public database, we designed oligos to amplify the COMT(bm3, wild type) gene from twelve diverse maize bm3 inbred lines and three non bm3 maize inbred lines.  The PCR products of the bm3 lines and non bm3 lines were cloned and sequenced.  An endpoint TaqMan PCR based zygosity assay was then developed to specifically detect and to test the zygosity status at the bm3 locus.  The assay utilizes a biplex of oligonucleotides specific to the bm3 deletion at the 3’ end of the exon and to the corresponding wild type sequences in the same assay.  Zygosity is determined by the presence/absence of the bm3 mutant and the wild type COMT alleles.  This high throughput PCR based molecular characterization of bm3 mutants will greatly enhance the breeding process for BMR maize lines.

See more from this Division: C01 Crop Breeding & Genetics
See more from this Session: Use of Molecular Tools to Enhance Breeding Efforts

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