Development of a multiplex amplicon‐sequencing assay to detect low‐frequency mutations in poinsettia ( <i>Euphorbia pulcherrima</i> ) breeding programmes

Poinsettia is an economically important ornamental potted plant in which certain bract colour variants are often obtained by mutation breeding. Previously, poinsettia, we identified Bract1, a GST gene involved the sequestration and transport of anthocyanins to vacuole. This carries short, highly mutable 4-bp repeat its coding region. Loss one unit leads loss function for homozygous mutants, anthocyanin-based coloration absent, resulting white or cream-coloured bracts. Although induction through ionizing radiation high frequency mutations mutants difficult obtain from red genotypes. In this study, used Bract1-specific amplicon sequencing as tool identify pools tissues, enabled detection dilutions up mutant 50 nonmutated samples. approach efficient screening recalcitrant genotypes mutated alleles reduction load application breeding programmes.