Bulk Segregant Analysis Technique to Target Seasonality and Runnering Loci in Diploid Strawberries

Bulked segregant analysis was used to determine inter-simple sequence repeat DNA (ISSR) markers for two genes, one controls seasonality and other runnering in diploid Fragaria spp. ISSR- PCR analysis was performed using these DNA pools as template. The bulks are screened for differences using anchored simple sequence polymorphic DNA primers which generates clean band profiles and polymorphic fragments were detected. Using 23 primers, either singly or pairwise, we established 16 markers tightly linked to the seasonality locus and 7 markers for runnering locus. This bulked segregant method can be applied for any species and any region of interest in which detailed linkage maps or physical maps are needed.

___

  • E. Hyams, Origins of the strawberry, its introduction into cultivation and
  • improvement. Chapter 2, p16-28 in: Strawberry Cultivation, (1953) Faber and Faber, London.
  • H. Hirsalmi, The role of natural small fruits in Finnish plant breeding Aquilo Ser Botanica, (1993). 31: 59-67.
  • T, Brown, P.F. Wareing, The genetical control of the everbearing habit and three other characters in varieties of Fragaria vesca. Euphytica (1965). 14: 97-112.
  • N.H. Battey, P. Le. Miere, A. Tehranifar, C. Cekic, S. Taylor, K.J. Shrives, P. Hadley, A.J. Greenland, J. Darby, M.J. Wilkinson, Genetic and environmental control of flowering in strawberry. Genetic and environmental manipulation of horticultural crops (1998). CAB INTERNATIONAL, Wallingford, UK. 111- 131. Journal of New Results in Science 9 (2015) 92-98.
  • G.L. Wang, A.H. Paterson, Assessment of DNA pooling strategies for mapping of QTLs. Theoretical and Applied Genetics (1994). 88:355-361.
  • R.W. Michelmore, I. Paran, R.V. Kessel, Identification of markers linked to disease resistance genes by bulked segregant analysis: a rapid method to detection of markers in specific genomic regions using segregation populations. Proceeding The National Academy of Science of USA (1991). 88: 9828-9832.
  • N.A. Tinker, D.E. Mather, M.G. Fortin, Pooled DNA for linkage analysis: practical and statistical considerations. Genome (1994). 37: 6, 999-1004.
  • R.E. Durham, S.S. Korban, Evidence of gene introgression in apple using RAPD markers. Euphytica (1994). 79:1-2, 109-114.
  • S.J. Kemp, A.J. Teale, Randomly primed PCR amplification of pooled DNA reveals polymorphism in a ruminant repetitive DNA sequence which differentiates B. indicus and B. taurus. Animal Genetics (1994). 25: 2, 83-88.
  • S.D. Haley, P.N. Miklas, J.R. Stavely, J. Byrum, J.D. Kelly, Identification of RAPD markers linked to a major rust resistance gene block in common bean. Theoretical and Applied Genetics (1993). 86: 4, 505-512.
  • N.I. Naqvi, J.M. Bonman, D.J. Mackill, R.J. Nelson, B.B. Chattoo, Identification of RAPD markers linked to a major blast resistance gene in rice Molecular Breeding (1995). 1: 4, 341-348.
  • H.M. Wain, A.A. Toye, S. Hughes, N. Bumstead, Targeting of marker loci to chicken chromosome 16 by representational difference analysis.Animal Genetics (1998). 29: 6, 446-452.
  • F. Gallego, B. Calles, C. Benito, Molecular markers linked to the aluminium tolerance gene Alt1 in rye (Secale cereale L.). Theoretical and Applied Genetics (1998). 97: 7:1104-1109.
  • J.S. Gavora, R.W. Fairfull, B.F. Benkel, W.J. Cantwell, J.R. Chambers, Prediction of heterosis from DNA fingerprints in chickens. Genetics (1996). 144: 2, 777-784.
  • E. Zietkiewicz, A. Rafalski, D. Labunda, Genome fingerprinting by simple sequence repeat (SSR)-anchored polymerase chain reaction amplification. Genomics (1994). 20:176-183.
  • C. Cekic, N.H. Battey M.J. Wilkinson, The potential of ISSR-PCR Primer Pair Combinations for Genetic Linkage Analysis using the Seasonal Flowering Locus in Fragaria vesca as a Model. Theoretical and Applied Genetics (2001). 103:4, 540-546.
  • K. Wu, R. Jones, L. Danneberger, P.A. Scolnik, Detection of microsatellite polymorphisms without cloning. Nucleic Acids Research (1994). 22:3257-3258.
  • S. Moreno, J.P. Martin, J.M. Ortiz, Inter-simple sequence repeats PCR for characterisation of closely related grapevine germplasm. Euphytica (1998). 101:117-125.
  • J.G.K. Williams, J.A. Hanafey, J.A. Rafalski, S.V. Tingey, Genetic analysis using random amplified polymorphic DNA markers. Methods of Enzymology (1993). 218:704-740.
  • J.J. Doyle, J.L. Doyle, A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochemical Bulletin (1987). 19:11-15.
  • Y.M. Charters, A. Robertson, M.J. Wilkinson, G., Ramsay, PCR analysis of oilseed rape cultivars (Brassica napus L ssp oleifera) using 5'-anchored simple sequence repeat (SSR) primers. Theoretical and Applied Genetics. (1996). 92:3-4,442- 447.