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Development and characterization of simple sequence repeats for flowering dogwood (Cornus florida L.)

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Abstract

Abundant, codominant simple sequence repeats (SSRs) markers can be used for constructing genetic linkage maps and in marker-assisted breeding programs. Enrichment methods for SSR motifs were optimized with the ultimate aim of developing numerous loci in flowering dogwood (C. florida L.) genome. Small insert libraries using four motifs (GT, CT, TGG, and AAC) were constructed with C. florida ‘Cherokee Brave’ deoxyribonucleic acid (DNA). Colony polymerase chain reaction (PCR) of 2,208 selected clones with three primers we reported previously indicated that 47% or 1,034 of the clones harbored one of the four targeted SSR motifs. Sequencing the putative positive clones confirmed that nearly 99% (1,021 of 1,034) of them contained the desired motifs. Of the 871 unique SSR loci, 617 were dinucleotide repeats (70.8%), and 254 were trinucleotide or longer repeats (29.2%). In total, 379 SSR loci had perfect structure, 237 had interrupted, and 255 had compound structure. Primer pairs were designed from 351 unique sequences. The ability of the 351 SSR primer pairs to amplify specific loci was evaluated with genomic DNA of ‘Appalachian Spring’ and ‘Cherokee Brave’. Of these primers, 311 successfully amplified product(s) with ‘Cherokee Brave’ DNA, 21 produced weak or faint products, and 19 did not amplify any products. Additionally, 218 of the 311 primers pairs revealed polymorphisms between the two cultivars, and 20 out of 218 primers detected an average of 13.7 alleles from 38 selected Cornus species and hybrids. These SSR loci constitute a valuable resource of ideal markers for both genetic linkage mapping and gene tagging of flowering dogwood.

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Acknowledgments

This work was supported by USDA Agreement no. 58-6404-2-0057. We thank Mary Duke, Linda Ballard, and Xiaofen Liu for library sequencing. Any library/sequence information requirements can be addressed to Dr. Robert N. Trigiano at rtrigian@utk.edu. Mention of trade names or commercial products in this article is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the US Department of Agriculture.

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Authors and Affiliations

  1. Entomology and Plant Pathology, University of Tennessee, 2431 Joe Johnson Drive, Knoxville, TN, 37996, USA

    X. Wang, R. Trigiano & M. Windham

  2. MSA Genomics Laboratory, USDA-ARS-CGRU, 141 Experiment Station Rd., Stoneville, MS, 38776, USA

    B. Scheffler

  3. MSA Southern Horticultural Laboratory, USDA-ARS, 810 Highway 26 West, Poplarville, MS, 39470, USA

    T. Rinehart & J. Spiers

Authors
  1. X. Wang
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  2. R. Trigiano
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  3. M. Windham
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  4. B. Scheffler
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  5. T. Rinehart
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  6. J. Spiers
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Corresponding author

Correspondence to R. Trigiano.

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Communicated by M. Morgante

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Table S1

SSR primers developed for flowering dogwood (Cornus florida ) (DOC 697 KB)

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Wang, X., Trigiano, R., Windham, M. et al. Development and characterization of simple sequence repeats for flowering dogwood (Cornus florida L.). Tree Genetics & Genomes 4, 461–468 (2008). https://doi.org/10.1007/s11295-007-0123-z

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  • DOI: https://doi.org/10.1007/s11295-007-0123-z

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