A high-throughput genotyping system combining rapid DNA extraction and high-resolution melting analysis in allo-octoploid strawberry
The strawberry is an important commodity as well as a model plant species in the Rosaceae family. DNA markers for important traits in octoploid strawberry have increased dramatically in recent years, and are now widely applied for marker-assisted selection (MAS) to assist strawberry breeding. To date, laborious and expensive DNA extraction procedures have been used to achieve sufficient DNA quality in this recalcitrant species. When combined with gel-based marker systems, current genotyping methods are becoming a bottleneck for MAS in large breeding populations. Here we present a high-throughput genotyping system for octoploid strawberry as a model for other Rosaceae fruit crop breeding programs. High-resolution melting (HRM) analysis was combined with rapid DNA extraction for selection at the FaFAD1, FaRPc1 and FaRXf1 loci conferring fruity-aroma (γ-decalactone), resistance to Phytophthora crown root rot (Phytophthora cactorum) and bacterial angular leaf spot (Xanthamonas fragariae), respectively. This system was successfully employed to screen approximately 8,000 seedlings in the University of Florida strawberry breeding program in 2015. The system is rapid, cost-effective and highly accurate and should be readily applicable to other Rosaceae fruit crops with minor modifications. Updates and insights from seedling screening in spring, 2016 will also be discussed.
Lee, S., Noh, Y.-H., Roach, J.A., Mangandi, J., Verma, S., Whitaker, V.M. and Cearley, K.R. (2017). A high-throughput genotyping system combining rapid DNA extraction and high-resolution melting analysis in allo-octoploid strawberry. Acta Hortic. 1156, 89-94
γ-decalactone, angular leaf spot, Phytophthora crown root rot, single nucleotide polymorphism (SNP), marker-assisted selection (MAS), high-resolution melting (HRM) methods for genotyping