High-throughput DNA extraction and allele specific PCR primers enables efficient screening for mutant (gsf) and wild-type (GSF) alleles in eastern gamagrass

Abstract

Eastern gamagrass [Tripsacum dactyloides (L.) L.], is a perennial warm-season bunchgrass with high forage potential if properly managed. Low seed production is one factor that has limited wide spread use of this native grass. A recessive gynomonoecious sex form (GSF) mutation previously found in a wild population has the potential to increase seed production, although it may also be linked to other deleterious traits. Allele specific primers were designed to detect the wild-type (GSF) and mutant (gsf) allele. Primers were successful at confirming the genotype of an assortment of plants tested using template DNA from a high-throughput extraction protocol. A tissue-pooling strategy enabled detection of the mutant allele when present in a predominantly wild-type background. The low cost, lack of toxic waste, and speed of the DNA extraction protocol and the ability to efficiently genotype a large number of individuals makes this a practical tool in a breeding program working with the GSF trait. © Crop Science Society of America.