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.
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Anderson JA, Ogihara Y, Sorrells ME, Tanksley SD (1992) Development of a chromosomal arm map for wheat based on RFLP markers. Theor Appl Genet 83:1035–1043
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 78:495–504
Devos KM, Gale MD (1992) The use of randomly amplified DNA markers in wheat. Theor Appl Genet 84:567–572
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 15:169–171
Dweikat I, MacKenzie S, Levy M, Ohm H (1993) Pedigree assessment using RAPD-DGGE in cereal crop species. Theor Appl Genet 83:497–505
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 34:362–374
He S, Ohm H, MacKenzie S (1992) Detection of DNA sequence polymorphisms among wheat varieties. Theor Appl Genet 84:576–578
Kam-Morgan LNW, Gill BS, Muthukrishnan S (1989) DNA restriction fragment length polymorphism: a strategy for genetic mapping the D genome of wheat. Genome 32:724–732
Olson M, Hood L, Cantor C, Dotstein D (1989) A common language for physical mapping of the human genome. Science 254:1434–1435
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 31:50–53
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 13–20
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 230:1350–1354
Sanger F, Niclen S, Coulsen AR (1977) DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Sears ER (1954) The aneuploids of common wheat. Res Bull Mo Agric Exp Stn 472
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 78:342–348
Talbert LE, Moylan SL, Hansen LJ (1992) Assessment of repetitive DNA variation among accessions of hexaploid and tetraploid wheat. Crop Sci 32:366–369
Tragoonrung S, Kanazin V, Hayes PM, Blake TK (1992) Sequence-tagged-site-facilitated PCR for barley genome mapping. Theor Appl Genet 84:1002–1008
Weining S, Langridge P (1991) Identification and mapping of polymorphisms in cereals based on the polymerase chain reaction. Theor Appl Genet 82:209–216
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 18:6531–6535
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Communicated by G. E. Hart
Contribution no. J-2833 of the Montana Agric Exp Stn
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Talbert, L.E., Blake, N.K., Chee, P.W. et al. Evaluation of “sequence-tagged-site” PCR products as molecular markers in wheat. Theoret. Appl. Genetics 87, 789–794 (1994). https://doi.org/10.1007/BF00221130
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DOI: https://doi.org/10.1007/BF00221130