High Resolution Melt Analysis of Simple Sequence Repeats for Bentgrass Species Differentiation

Agrostis stolonifera L. (creeping bentgrass) and Agrostis capillaris (colonial bentgrass) are turfgrass species well adapted for golf course use in regions of the world where cool-season grasses are grown. Interspecific hybrids between the species do form and have the potential to incorporate some of the beneficial characteristics of both species. A. stolonifera has excellent quality at low mowing heights and recovers well from damage. A. capillaris tends to exhibit more drought tolerance and a higher level of resistance to the common fungal pathogen Sclerotinia homoeocarpa. Fast inexpensive methods of species differentiation could help seed certification agencies and enhance interspecific hybrid bentgrass development. Ninety-six Simple Sequence Repeat (SSR) primer pairs were designed from Roche 454 sequencing of A. stolonifera and A. capillaris genomic DNA and were tested for their potential for bentgrass species differentiation. Real-time Polymerase Chain Reaction (PCR) followed by High Resolution Melt Analysis (HRM) was tested for its potential to speed up analysis and lower costs. SSR primer pairs were identified that can differentiate bentgrass species.