Theoretical and Applied Genetics

, Volume 87, Issue 7, pp 789–794 | Cite as

Evaluation of “sequence-tagged-site” PCR products as molecular markers in wheat

  • L. E. Talbert
  • N. K. Blake
  • P. W. Chee
  • T. K. Blake
  • G. M. Magyar
Article

Abstract

The polymerase chain reaction (PCR) is an attractive technique for many genome mapping and characterization projects. One PCR approach which has been evaluated involves the use of randomly amplified polymorphic DNA (RAPD). An alternative to RAPDs is the sequence-tagged-site (STS) approach, whereby PCR primers are designed from mapped low-copy-number sequences. In this study, we sequenced and designed primers from 22 wheat RFLP clones in addition to testing 15 primer sets that had been previously used to amplify DNA sequences in the barley genome. Our results indicated that most of the primers amplified sequences that mapped to the expected chromosomes in wheat. Additionally, 9 of 16 primer sets tested revealed polymorphisms among 20 hexaploid wheat genotypes when PCR products were digested with restriction enzymes. These results suggest that the STS-based PCR analysis will be useful for generation of informative molecular markers in hexaploid wheat.

Key words

Wheat Molecular Markers Breeding 

References

  1. Anderson JA, Ogihara Y, Sorrells ME, Tanksley SD (1992) Development of a chromosomal arm map for wheat based on RFLP markers. Theor Appl Genet 83:1035–1043Google Scholar
  2. Chao S, Sharp PJ, Worland AJ, Warham EJ, Koebner RMD, Gale MD (1989) RFLP-based genetic maps of wheat homoeologous group 7 chromosomes. Theor Appl Genet 78:495–504Google Scholar
  3. Devos KM, Gale MD (1992) The use of randomly amplified DNA markers in wheat. Theor Appl Genet 84:567–572Google Scholar
  4. D'Ovidio R, Tanzarella OA, Porceddu E (1990) Rapid and efficient detection of genetic polymorphisms in wheat through amplification by the polymerase chain reaction. Plant Mol Biol 15:169–171Google Scholar
  5. Dweikat I, MacKenzie S, Levy M, Ohm H (1993) Pedigree assessment using RAPD-DGGE in cereal crop species. Theor Appl Genet 83:497–505Google Scholar
  6. Gill KS, Lubbers EL, Gill BS, Raupp WJ, Cox TS (1991) A genetic linkage map of Triticum tauschii (DD) and its relationship to the D genome of bread wheat (AABBDD). Genome 34:362–374Google Scholar
  7. He S, Ohm H, MacKenzie S (1992) Detection of DNA sequence polymorphisms among wheat varieties. Theor Appl Genet 84:576–578Google Scholar
  8. Kam-Morgan LNW, Gill BS, Muthukrishnan S (1989) DNA restriction fragment length polymorphism: a strategy for genetic mapping the D genome of wheat. Genome 32:724–732Google Scholar
  9. Olson M, Hood L, Cantor C, Dotstein D (1989) A common language for physical mapping of the human genome. Science 254:1434–1435Google Scholar
  10. Quick JS, Nkongolo KK, Meyer WL, Peairs FB, Weaver B (1991) Russian wheat aphid reaction and agronomic and quality traits of resistant wheats. Crop Sci 31:50–53Google Scholar
  11. Saiki RK (1990) Amplification of genomic DNA. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ (eds) PCR protocols: a guide to methods and applications. Academic Press, New York, pp 13–20Google Scholar
  12. Saiki RK, Scarf S, Falloona F, Mullis KB, Horn GT, Erlich HA, Arnheim N (1985) Enzymatic amplification of beta-globulin genomic sequences and restriction analysis for diagnosis of sickle cell anemia. Science 230:1350–1354Google Scholar
  13. Sanger F, Niclen S, Coulsen AR (1977) DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467PubMedGoogle Scholar
  14. Sears ER (1954) The aneuploids of common wheat. Res Bull Mo Agric Exp Stn 472Google Scholar
  15. Sharp PJ, Chao S, Desai S, Gale MD (1989) The isolation, characterization and application in the Triticeae of a set of wheat RFLP clones identifying each homoeologous arm. Theor Appl Genet 78:342–348Google Scholar
  16. Talbert LE, Moylan SL, Hansen LJ (1992) Assessment of repetitive DNA variation among accessions of hexaploid and tetraploid wheat. Crop Sci 32:366–369Google Scholar
  17. Tragoonrung S, Kanazin V, Hayes PM, Blake TK (1992) Sequence-tagged-site-facilitated PCR for barley genome mapping. Theor Appl Genet 84:1002–1008Google Scholar
  18. Weining S, Langridge P (1991) Identification and mapping of polymorphisms in cereals based on the polymerase chain reaction. Theor Appl Genet 82:209–216Google Scholar
  19. Williams JGK, Kubelik ARK, Livak JL, Rafalslki JA, Tingey SV (1990) DNA polymorphisms amplified by random primers are useful as genetic markers. Nucleic Acids Res 18:6531–6535PubMedGoogle Scholar

Copyright information

© Springer-Verlag 1994

Authors and Affiliations

  • L. E. Talbert
    • 1
  • N. K. Blake
    • 1
  • P. W. Chee
    • 1
  • T. K. Blake
    • 1
  • G. M. Magyar
    • 1
  1. 1.Department of Plant and Soil ScienceMontana State UniversityBozemanUSA

Personalised recommendations