B.E. Sayed-Tabatabaei^{1, 3}, T. Komatsuda¹, F. Takaiwa¹, A. Graner²

1. Department of Biotechnology, National Institute of Agrobiological Resources, Kannondai, Tsukuba, Ibaraki 305, Japan

2. Institute for Plant Genetics and Crop Plant Research, Corrensstr 3, D-06466 Gatersleben, Germany

3. Present address: Department of Plant Breeding, College of Agriculture, University of Tehran, Karaj, Iran

Conversion of RFLP clone's DNA sequences to oligonucleotide primers for polymerase chain reaction can lessen the time and expenses of plant genome mapping experiments. Their amplified DNA products (sequenced-tagged-sites) identify known locations on chromosomes. A series of barley markers amplified by primer sets developed from ABC and ABG clones (Kleinhofs et al. 1993) have been described previously (Blake et al. 1996). Another series of markers amplified by primer sets developed from MWG and cMWG clones (Graner et al. 1991, 1994, A Graner, unpublished) have been used for the integration of translocation breakpoints (Sorokin et al. 1995, Korzun and Künzel 1996, 1997). We sequenced the MWG and cMWG clones evenly distributed in the barley genome and designed the primer pairs.

Double-strand sequencing of plasmid was performed using ABI fluorescent dye terminator system and automated DNA sequencer (373A). Primer sequences were selected using Oligo software. The designed primers consisted of 25 nucleotides to permit annealing temperature of 42 to 66deg.C which allowed the amplification of single DNA fragment from most primers (Table 1). Chromosomal location of the amplified DNA fragments and the constructed linkage map will be published elsewhere (Mano et al. in preparation). Clone and primer sequences are deposited in Graingenes. Small aliquots of all the primer sets in Table 1 are available from T. Komatsuda.

Acknowledgments: The research grant from the Science and Technology Agency (Enhancement of Center of Excellence, Special Coordination Funds for Promoting Science and Technology) to F. T. is greatly appreciated.

References

Blake TK, Kadyrzhanova D, Shepherd KW, Islam AKMR, Langridge PL, McDonald CL, Erpelding J, Larson S, Blake NK, Talbert LE (1996) STS-PCR markers appropriate for wheat-barley introgression. Theor Appl Genet 93:826-832.

Graner A, Jahoor A, Schondelmaier J, Siedler H, Pillen K, Fischbeck G, Wenzel G, Herrmann RG (1991) Construction of an RFLP map of barley. Theor Appl Genet 83:250-256.

Graner A, Bauer E, Kellermann A, Kirchner S, Muraya JK, Jahoor A, Wenzel G (1994) Progress of RFLP-map construction in winter barley. BGN 23:53-593.

Kleinhofs A, Kilian A, Saghai Maroof MA, Biyashev RM, Hayes P, Chen FQ, Lapitan N, Fenwick A, Blake TK, Kanazin V, Ananiev E, Dahleen L, Kudrna D, Bollinger J, Knapp SJ, Liu B, Sorrells M, Heun M, Franckowiak JD, Hoffman D, Skadsen R, Steffenson BJ (1993) A molecular, isozyme and morphological map of the barley (Hordeum vulgare) genome. Theor Appl Genet 86:705-712.

Korzun L, Künzel G (1996) Integration of translocation breakpoints of barley chromosome 3 and 7 into the Igri/Franka derived RFLP map. BGN 25:13-16.

Korzun L, Künzel G (1997) Integration of 70 translocation breakpoints into the Igri/Franka-derived RFLP maps for 2H and 6H. BGN 25:13-16.

Sorokin A, Marthe F, Künzel G (1995) Integration of 37 translocation breakpoints of barley chromosome 5 into the Igri/Franka derived RFLP maps. BGN 24:108-112.