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Title: Tall Fescue Genomic SSR Markers: Development and Transferability across Multiple Grass Species

Author
item SAHA, MALAY - SAMUEL ROBERTS NOBLE FOUN
item COOPER, JOHN - SAMUEL ROBERTS NOBLE FOUN
item Mian, Rouf
item CHEKHOVSKIY, KONSTANTIN - SAMUEL ROBERTS NOBLE FOUN
item MAY, GREGORY - SAMUEL ROBERTS NOBLE FOUN

Submitted to: Theoretical and Applied Genetics
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 8/4/2006
Publication Date: 9/1/2006
Citation: Saha, M.C., Cooper, J.D., Mian, R.M., Chekhovskiy, K., May, G.D. 2006. Tall Fescue Genomic SSR Markers: Development and Transferability across Multiple Grass Species. Theoretical and Applied Genetics. 113:1449-1458.

Interpretive Summary: Simple sequence repeat (SSR) marker has become a very useful tool for modern plant breeding and genetic research. Tall fescue (Festuca arundinacea Schreb) is major cool season forage and turf grass in the USA as well as in other temperate regions of the world. Currently, few SSR markers are available for tall fescue and other forage grass species. The objectives of this study were to develop SSR primer pairs (PPs) from tall fescue genomic DNA and assess their potential usefulness in different forage, cereal, and turf grass species. A total of 511 PPs were developed and their usefulness was tested across six different grass species (tall fescue, meadow fescue, perennial ryegrass, Italian ryegrass, rice and wheat). Results revealed that 48% (in rice) to 66% (in tall fescue) of the PPs produced clean SSR-type amplification products in different grass species. Polymorphism rates were fairly high in tall fescue (68%) and substantially lower in other species (46% ryegrass, 39% wheat, and 34% rice). A set of SSR loci (38%) were identified which amplified across all six species. These markers were also tested for construction of a genetic linkage map in tall fescue and were found highly useful for this purpose. Sequencing of amplified products obtained from primer pair NFFAG428 revealed a very high level of sequence similarity among the grass species under study. The results of this study demonstrate the usefulness of these SSRs for genetic linkage mapping in tall fescue and cross-species amplification. All of the genomic SSR PPs developed in this study are being made publicly available through this publication. This will provide the forage and turf grass community with a large set of SSR markers for the first time.

Technical Abstract: Simple sequence repeat (SSR) markers are highly informative and widely used for genetic and breeding studies. Currently, a very limited number of SSR markers are available for tall fescue (Festuca arundinacea Schreb) and other forage grass species. A tall fescue genomic library enriched in (GA/CT)n repeats was used to develop SSR markers. Our objectives were to develop SSR primer pairs (PPs) from tall fescue and assess PP-functionality across different forage, cereal, and turf grass species. A total of 511 PPs were developed and their utility was determined across six different grass species. The parents and a subset of a tall fescue mapping population were used to select PPs for mapping in tall fescue. Results revealed that 48% (in rice) to 66% (in tall fescue) of the PPs produced clean SSR-type amplification products in different grass species. Polymorphism rates were fairly high in tall fescue (68%) and substantially lower in other species (46% ryegrass, 39% wheat, and 34% rice). A set of SSR loci (38%) were identified which amplified across all six species. Loci segregating in the tall fescue mapping population were grouped as loci segregating from the female parent (HD28-56, 37%), the male parent (R43-64, 37%), and both parents (26%). Three percent of the loci that were polymorphic between parents were monomorphic in the F1 mapping population and the remaining loci segregated. Sequencing of amplified products obtained from primer pair NFFAG428 revealed a very high level of sequence similarity among the grass species under study. Our results are the first report of genomic SSR marker development from tall fescue and the outcomes demonstrate the usefulness of these SSRs for genetic linkage mapping in tall fescue and cross-species amplification.