ISSN: 2456-3102
Jocelyn De Wever
Plant genetic diversity studies are of high importance for efficient plant conversation and resource strategies (eg. tackling mislabeling, conserving valuable genetic material, parentage analysis, and genetic diversity studies) as they contribute to an increased knowledge on the genetic background and diversity of specific plants. These studies are most commonly analyzed through simple and effective genotyping methods making use of genetic markers, such as SSRs, however SNPs are gaining more interest. Recently, a cost-effective qPCRbased method has been proposed for SNP genotyping purposes, coined doublemismatch allele-specific (DMAS) qPCR as cheap alternative to -sample and multilocus method, based on straightforward readout of DNAbinding dye based qPCR technology. Its design, optimization, validation and application on Theobroma cacao L., an important cash crop involved in the chocolate production, has shown successful. It offered valuable knowledge on the background of cocoa which is often plagued by mislabeling and inefficient and limited management resources. The method, optimized here, showed 98.05% efficient in calling the right cacao genotype and identified 15.38% off-types and two duplicates in an internationally recognized cacao population (n=65), using a limited amount of markers (n=42). Furthermore, only 13 markers were needed to differentiate all analyzed accessions. Notably, the described method can easily be optimized and implemented in any molecular biology lab for a wide range of objectives and organisms e.g. mutation detection and to facilitate gene mapping and markerassisted selection for breeding purposes.