Detection of interspecific hybrids between creeping bentgrass and colonial bentgrass using DNA marker analysis

Abstract Bentgrasses (Agrostis spp.) are cool‐season turfgrasses that have utility on golf courses or other turf applications requiring low mowing heights. Interspecific hybridization of Agrostis species is a strategy that has been proposed to enhance biotic and abiotic stress tolerance in this genus. Non‐transgenic, efficient, and cost‐effective methods for interspecific hybrid detection are needed. In this study two bentgrass species creeping bentgrass (A. stolonifera L.), and colonial bentgrass (A. capillaris L.) were allowed to intercross under field conditions. Progeny plant DNA was extracted and stored using Flinders Technology Associates (FTA) PlantSaver cards followed by primer specific polymerase chain reaction (PCR) and high‐resolution melt analysis. Species identifying PCR primers were optimized for real‐time PCR through fragment resequencing of species‐specific sequence characterized amplified region (SCAR) markers. A total of 2289 potential interspecific hybrid bentgrass progeny were screened with 99.4% of the DNA extractions successful. The percent interspecific hybridization, on an individual plant basis, ranged from 1 to 85% and a total of 524 interspecific‐hybrid bentgrasses were identified representing 23% of plants tested. Interspecific hybridization is common between these bentgrass species and the proposed detection method provides a non‐transgenic, efficient, and cost‐effective method for interspecific hybrid identification.