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Research Project: Sustainable Approaches for Pest Management in Vegetable Crops

Location: Vegetable Research

Title: Evaluation of off-type grasses in hybrid bermudagrass (Cynodon dactylon (L.) Pers. x C. transvaalensis Burtt-Davy) putting greens using genotyping-by-sequencing

item REASOR, ERIC - University Of Tennessee
item BROSNAN, JAMES - University Of Tennessee
item STATON, MARGARET - University Of Tennessee
item LANE, THOMAS - University Of Tennessee
item TRIGIANO, ROBERT - University Of Tennessee
item Wadl, Phillip
item CONNER, JOANN - University Of Georgia
item SCHWARTZ, BRIAN - University Of Georgia

Submitted to: American Society of Agronomy
Publication Type: Abstract Only
Publication Acceptance Date: 6/15/2016
Publication Date: 11/7/2016
Citation: Reasor, E., Brosnan, J.T., Staton, M.E., Lane, T., Trigiano, R., Wadl, P.A., Conner, J.A., Schwartz, B.M. 2016. Evaluation of off-type grasses in hybrid bermudagrass (Cynodon dactylon (L.) Pers. x C. transvaalensis Burtt-Davy) putting greens using genotyping-by-sequencing. American Society of Agronomy. Abstract ID: 99766.

Interpretive Summary: N/A

Technical Abstract: Use of hybrid ultradwarf bermudagrasses (UDBG; Cynodon dactylon (L.) Pers. x C. transvaalensis Burtt-Davy) on golf course putting greens is increasing in the southern United States. However, off-type grasses within many putting surfaces have been observed. To explore the genetic variation among UDBG cultivars and off-type grasses, a genotyping-by-sequencing (GBS) approach was utilized. Desirable UDBG cultivars and off-type grasses were selected from golf course-putting greens in 2013, and cultured from single stolon transplants in a glasshouse at the University of Tennessee (Knoxville, TN). All hybrid bermudagrass and off-type samples were confirmed as triploids (2n=3x=27) using flow cytometry. A GBS protocol was then used to assess the genetic variation among UDBG cultivars, off-types, and progenitor species. The ApeKI restriction enzyme was selected based on optimization trials for the GBS digestion in order to maximize the number of sampled genomic loci. Libraries were constructed from 96 DNA samples and multiplexed on a single lane of an Illumina HiSeq. Over 270 million reads were obtained. Data analysis was performed with the UNEAK pipeline designed for calling single nucleotide polymorphisms (SNPs) from GBS data without a reference genome. Using a minimum depth of 3 reads per sample, the analysis yielded 205,416 filtered SNPs (average read depth of 3.889). The samples were run on a lane of an Illumina NextSeq to increase coverage. Analysis of the combined datasets yielded 272,039 filtered SNPs with an average depth of 4.718. A multidimensional scaling plot of the genetic variation among samples revealed clear clustering of the progenitor species (C. dactylon (L.) Pers. and C. transvaalensis Burtt-Davy) from UDBG cultivars and off-types. Of the 26 off-type samples, only 9 (35%) were genetically divergent from UBDG cultivars. Additional research is needed to determine the genetic origins of off-type grass phenotypes in UDBG putting greens.