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Implementation of simple sequence repeat markers to genotype Florida strawberry varieties

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Abstract

Simple sequence repeats (SSRs or microsatellites) are frequently used as a robust, rapid, and relatively inexpensive means of genotyping. Recent reports in cultivated strawberry (Fragaria × ananassa) have identified a widely applicable set of SSRs that permits discrimination of closely-related genotypes. In the present study this same set of SSRs is analyzed in cultivars commonly grown in the state of Florida, as well as advanced breeding selections from the University of Florida program. Nine primer pairs have been used to produce discrete SSR patterns for all lines tested, including diagnostic sets that distinguish between closely-related cultivars and/or breeding selections. A comparison of markers to known pedigrees is presented, along with analysis of relative relatedness between lines studied. We also detail important technical considerations for transferability and limitations of these technologies between laboratories. The resulting genotype data for cultivars commonly used in Florida are accessible as supplemental data in graphic format and allow comparison with other cultivars. Users can thus select the most diagnostic SSRs to test the lines in question. These resources provide new tools for breeders and nurseries to authenticate genotypes, follow inheritance patterns and enforce patent protection. As important, this report underscores the strengths and weaknesses of applying the original methodologies to a different plant population in independent laboratories.

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References

  • Alba V, Montemurro C, Sabetta W, Pasqualone A, Blanco A (2009) SSR-based identification key of cultivar of Olea europaea L. diffused in Southern-Italy. Sci Hortic 123:11–16

    Article  CAS  Google Scholar 

  • Arnau G, Lallemand J, Bourgoin M (2003) Fast and reliable strawberry cultivar identification using inter simple sequence repeat (ISSR) amplification. Euphytica 129:69–79

    Article  CAS  Google Scholar 

  • Ashley MV, Wilk JA, Styan SMN, Craft KJ, Jones KL, Feldheim KA, Lewers KS, Ashman TL (2003) High variability and disomic segregation of microsatellites in the octoploid Fragaria virginiana Mill. (Rosaceae). Theor Appl Genet 107:1201–1207

    Article  CAS  PubMed  Google Scholar 

  • Bassil NV, Gunn M, Folta K, Lewers K (2006) Microsatellite markers for Fragaria from ‘Strawberry Festival’ expressed sequence tags. Mol Ecol Notes 6:473–476

    Article  CAS  Google Scholar 

  • Buteler MI, Jarret RL, LaBonte DR (1999) Sequence characterization of microsatellites in diploid and polyploid Ipomoea. Theor Appl Genet 99:123–132

    Article  CAS  Google Scholar 

  • Chang S, Puryear J, Cairney J (1993) A simple and efficient method for isolating RNA from pine trees. Plant Mol Biol Rep 11:113–116

    Article  CAS  Google Scholar 

  • Cipriani G, Pinosa F, Bonoli M, Faedi W (2006) A new set of microsatellite markers for Fragaria species and their application in linkage analysis. J Hortic Sci Biotechnol 81:668–675

    CAS  Google Scholar 

  • Clark JM (1988) Novel non-templated nucleotide addition reactions catalyzed by procaryotic and eucaryotic DNA polymerases. Nucleic Acids Res 16:9677–9686

    Article  CAS  PubMed  Google Scholar 

  • Congiu L, Chicca M, Cella R, Rossi R, Bernacchia G (2000) The use of random amplified polymorphic DNA (RAPD) markers to identify strawberry varieties: a forensic application. Mol Ecol 9:229–232

    Article  CAS  PubMed  Google Scholar 

  • Dale A, Sjulin TM (1990) Few cytoplasms contribute to North-American strawberry cultivars. HortScience 25:1341–1342

    Google Scholar 

  • Dangl GS, Mendum ML, Prins BH, Walker MA, Meredith CP, Simon CJ (2001) Simple sequence repeat analysis of a clonally propagated species: a tool for managing a grape germplasm collection. Genome 44:432–438

    Article  CAS  Google Scholar 

  • Dangl GS, Woeste K, Aradhya MK, Koehmstedt A, Simon C, Potter D, Leslie CA, McGranahan G (2005) Characterization of 14 microsatellite markers for genetic analysis and cultivar identification of walnut. J Am Soc Hortic Sci 130:348–354

    CAS  Google Scholar 

  • Dangl GS, Lee EW, Sims ST, Golino DA (2007) A new system for strawberry cultivar identification developed at Foundation Plant Services (FPS), University of California, Davis, using simple sequence repeat (SR) primers. In: NASS/NASGA proceedings, pp 118–121

  • Dangl GS, Yang J, Golino D, Gradziel T (2009) A practical method for almond cultivar identification and parental analysis using simple sequence repeat markers. Euphytica 168:41–48

    Article  CAS  Google Scholar 

  • Degani C, Rowland L, Levi A, Hortynski J, Galletta G (1998) DNA fingerprinting of strawberry (Fragaria × ananassa) cultivars using randomly amplified polymorphic DNA (RAPD) markers. Euphytica 102:247–253

    Article  CAS  Google Scholar 

  • Ellsworth DL, Rittenhouse D, Honeycutt RL (1993) Artifactual variation in randomly amplified polymorphic DNA binding patterns. Biotechniques 14:214–217

    CAS  PubMed  Google Scholar 

  • Folta K, Dhingra A, Howard L, Stewart P, Chandler C (2006) Characterization of LF9, an octoploid strawberry genotype selected for rapid regeneration and transformation. Planta 224:1058–1067

    Article  CAS  PubMed  Google Scholar 

  • Garcia MG, Ontivero M, Diaz Ricci JC, Castagnaro A (2002) Morphological traits and high resolution RAPD markers for the identification of the main strawberry varieties cultivated in Argentina. Plant Breed 121:76–80

    Article  CAS  Google Scholar 

  • Garcia-Vallve S, Palau J, Romeu A (1999) Horizontal gene transfer in glycosyl hydrolases inferred from codon usage in Escherichia coli and Bacillus subtilis. Mol Biol Evol 16:1125–1134

    CAS  PubMed  Google Scholar 

  • Gil-Ariza DJ, Amaya I, Botella MA, Munoz BJ, Caballero JL, Lopez-Aranda JM, Valpuesta V, Sanchez-Sevilla JF (2006) EST-derived polymorphic microsatellites from cultivated strawberry (Fragaria × ananassa) are useful for diversity studies and varietal identification among Fragaria species. Mol Ecol Notes 6:1195–1197

    Article  CAS  Google Scholar 

  • Govan C, Simpson DW, Johnson AW, Tobutt KR, Sargent DJ (2008) A reliable multiplexed microsatellite set for genotyping Fragaria and its use in a survey of 60 F. × ananassa cultivars. Mol Breeding 22:649–661

    Article  CAS  Google Scholar 

  • Graham J, McNicol RJ, McNicol JW (1996) A comparison of methods for the estimation of genetic diversity in strawberry cultivars. Theor Appl Genet 93:402–406

    Article  CAS  Google Scholar 

  • Guilford P, Prakash S, Zhu JM, Rikkerink E, Gardiner S, Bassett H, Forster R (1997) Microsatellites in Malus × domestica (apple): abundance, polymorphism and cultivar identification. Theor Appl Genet 94:249–254

    Article  CAS  Google Scholar 

  • Gupta PK, Balyan IS, Sharma PC, Ramesth B (1996) Microsatellites in plants—a new class of molecular markers. Curr Sci 70:45–54

    CAS  Google Scholar 

  • Hancock JF, Callow PA, Shaw DV (1994) Randomly amplified polymorphic DNAs in the cultivated strawberry, Fragaria × ananassa. J Am Soc Hortic Sci 119:862–864

    CAS  Google Scholar 

  • Kuras A, Korbin M, Zurawicz E (2004) Comparison of suitability of RAPD and ISSR techniques for determination of strawberry (Fragaria × ananassa Duch.) relationship. Plant Cell Tissue Organ Cult 79:189–193

    Article  CAS  Google Scholar 

  • Lewers KS, Styan SMN, Hokanson SC, Bassil NV (2005) Strawberry genbank-derived and genomic simple sequence repeat (SSR) markers and their utility with strawberry, blackberry, and red and black raspberry. J Am Soc Hortic Sci 130:102–115

    CAS  Google Scholar 

  • Lian C, Nara K, Nakaya H, Zhou Z, Wu B, Miyashita N, Davis TM (2001) Development of microsatellite markers in polyploid Salix reinii. Mol Ecol Notes 1:160–161

    Article  CAS  Google Scholar 

  • MacPherson JM, Eckstein PE, Scoles GJ, Gajadhar AA (1993) Variability of the random amplified polymorphic DNA assay among thermal cyclers, and effects of primer and DNA concentration. Mol Cell Probes 7:293–299

    Article  CAS  PubMed  Google Scholar 

  • Marchese A, Tobutt KR, Caruso T (2005) Molecular characterisation of Sicilian Prunus persica cultivars using microsatellites. J Hortic Sci Biotechnol 80:121–129

    CAS  Google Scholar 

  • Milella L, Saluzzi D, Lapelosa M, Bertino G, Spada P, Greco I, Martelli G (2006) Relationships between an Italian strawberry ecotype and its ancestor using RAPD markers. Genet Resour Crop Evol 53:1715–1720

    Article  CAS  Google Scholar 

  • Monfort A, Vilanova S, Davis TM, Arús P (2006) A new set of polymorphic simple sequence repeat (SSR) markers from a wild strawberry (Fragaria vesca) are transferable to other diploid Fragaria species and to Fragaria × ananassa. Mol Ecol Notes 6:197–200

    Article  CAS  Google Scholar 

  • Morgante M, Olivieri AM (1993) PCR-amplified microsatellites as markers in plant genetics. Plant J 3:175–182

    Article  CAS  PubMed  Google Scholar 

  • Myakishev MV, Khripin Y, Hu S, Hamer DH (2001) High-throughput SNP genotyping by allele-specific PCR with universal energy-transfer-labeled primers. Genome Res 11:163–169

    Article  CAS  PubMed  Google Scholar 

  • Perez T, Albornoz J, Dominguez A (1998) An evaluation of RAPD fragment reproducibility and nature. Mol Ecol 7:1347–1357

    Article  CAS  PubMed  Google Scholar 

  • Rabouam C, Comes AM, Bretagnolle V, Humbert J, Perique G, Bigot Y (1999) Features of DNA fragments obtained by random amplified polymorphic DNA (RAPD) assays. Mol Ecol 8:493–503

    Article  CAS  PubMed  Google Scholar 

  • Sargent DJ, Hadonou AM, Simpson DW (2003) Development and characterization of polymorphic microsatellite markers from Fragaria viridis, a wild diploid strawberry. Mol Ecol Notes 3:550–552

    Article  CAS  Google Scholar 

  • Sargent DJ, Clarke J, Simpson DW, Tobutt KR, Arús P, Monfort A, Vilanova S, Denoyes-Rothan B, Rousseau M, Folta KM, Bassil NV, Battey NH (2006) An enhanced microsatellite map of diploid Fragaria. Theor Appl Genet 112:1349–1359

    Article  CAS  PubMed  Google Scholar 

  • Sarri V, Baldoni L, Porceddu A, Cultrera NGM, Frediani M, Belaj A, Trujillo I, Cionini PG (2006) Microsatellite markers are powerful tools for discriminating among olive cultivars and assigning them to geographically defined populations. Genome 49:1606–1615

    Article  CAS  PubMed  Google Scholar 

  • Slovin JP, Schmitt K, Folta KM (2009) An inbred line of the diploid strawberry Fragaria vesca f. semperflorens for genomic and molecular genetic studies in the Rosaceae. BMC Plant Methods 5:15

    Article  Google Scholar 

  • Tyrka M, Dziadczyk P, Hortyński J (2002) Simplified AFLP procedure as a tool for identification of strawberry cultivars and advanced breeding lines. Euphytica 125:273–280

    Article  CAS  Google Scholar 

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Acknowledgements

This research was funded by a grant from the University of Florida IFAS Dean for Research (KMF and NP). The authors thank Dr. Daniel Sargent for valuable discussions and Kayla Shea Childers for technical assistance. The authors also recognize Dr. Craig K. Chandler for helpful insights and starting materials.

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Correspondence to Kevin M. Folta.

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Brunings, A.M., Moyer, C., Peres, N. et al. Implementation of simple sequence repeat markers to genotype Florida strawberry varieties. Euphytica 173, 63–75 (2010). https://doi.org/10.1007/s10681-009-0112-4

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  • DOI: https://doi.org/10.1007/s10681-009-0112-4

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