Location: Floral and Nursery Plants ResearchTitle: Simple sequence repeat marker development and diversity analysis in buffalograss
|MARSHALL, COLLIN - University Of Nebraska|
|AMUNDSEN, KEENAN - University Of Nebraska|
Submitted to: Crop Science
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 1/24/2022
Publication Date: 4/26/2022
Citation: Marshall, C., Warnke, S.E., Amundsen, K. 2022. Simple sequence repeat marker development and diversity analysis in buffalograss. Crop Science. https://doi.org/10.1002/csc2.20725.
Interpretive Summary: Buffalograss is a low-input turfgrass species native to the central Great Plains of the United States. It is valuable as a turfgrass because of its relative low water use, minimal fertility requirements, pest resistance, rapid establishment, and infrequent mowing requirements. Molecular markers would be a useful tool to characterize buffalograss genetic resources and to guide breeding strategies. Scientists from the University of Nebraska and ARS developed DNA markers for buffalograss and used them to characterize a selected population of buffalograss plants. Ninety-six DNA markers were initially selected and tested on a panel of eight plants. Twenty-four of the ninety-six markers worked well and were used to screen the full population of plants. The buffalograss-specific markers will be useful for studying the variability of buffalograss, maintaining purity in variety development, and can support plant breeding strategies.
Technical Abstract: Buffalograss [Buchloë dactyloides (Nutt.) Engelm. syn. Bouteloua dactyloides (Nutt.) Columbus]is a stoloniferous low-input turfgrass species native to the central Great Plains of the United States. Several molecular marker techniques have been used to compare relationships among elite buffalograss germplasm, but most rely on random amplification, are transferred from other species, or are developed from limited DNA sequence information. The objectives of this study were to develop buffalograss derived simple sequence repeat markers (SSRs) from buffalograss sequence data using an in-silico bioinformatics pipeline, and use high resolution melt (HRM) analysis to screen the SSRs on buffalograss cultivars and germplasm. Transcriptomes from buffalograss cultivars ‘378’ and ‘Prestige’ were mined for SSRs using MISA. There were 259 conserved SSRs that were polymorphic between ‘378’ and ‘Prestige’. Of those, 96 were tested by HRM on a panel of eight genotypes and 24 were selected for producing melt profiles that could distinguish the genotypes. The selected reactions produced 356 distinct alleles when tested on a panel of 96 genotypes, including 88 buffalograss germplasm selections, six buffalograss cultivars, and two blue grama [Bouteloua gracilis (H. B. K.) Lag. ex Steud.] selections, with allele frequencies ranging from 0.01 to 0.97. A subset of 86 markers segregating among the blue grama and elite buffalograss cultivars distinguished each genotype and could be used as a marker panel for variety discrimination and maintaining genetic purity. The buffalograss-specific SSR markers presented here will be useful to study buffalograss genetic diversity, line purity maintenance, and for marker supported plant breeding strategies.