Journal of Applied Genetics

, Volume 49, Issue 2, pp 127–134 | Cite as

A new diagnostic SSR marker for selection of theRym4/Rym5 locus in barley breeding

  • Mirosław Tyrka
  • Dragan Perovic
  • Agnieszka Wardyńska
  • Frank Ordon
Original Article


Genomic sequence AY661558, representing a part of the BAC contig of theRym4/Rym5 locus conferring resistance to the barley yellow mosaic virus complex (BaMMV/BaYMV), was exploited in order to develop SSR markers for practical barley breeding. Out of 57 SSR motifs found within this sequence, primers were designed and tested for the 5 SSRs with the highest repeat length. The polymorphic SSR marker QLB1 co-segregated withrym4 andrym5 phenotypes in respective high-resolution mapping populations developed for the construction of the original BAC contig. The primers targeted 2 sites located 756 bp and 5173 bp downstream of the translation initiation factor 4E (Hv-eIF4E). Physical linkage of the QLB1 marker to theRym4/Rym5 locus was confirmed experimentally on Morex BAC 519J14, a seed BAC ofHv-eIF4E, and BAC 801A11, which is located proximally toHv-eIF4E. QLB1 revealed 7 alleles in a set of 100 winter barley lines and cultivars. Five alleles were found within 673 advanced breeding lines derived from applied Polish winter barley breeding programmes, which corresponds to a PIC value of 0.684. No recombinants betweenRym4/5 and QLB1 were detected, suggesting that QLB1 can be used efficiently in marker-assisted selection of theHv-eIF4E-mediated bymovirus resistance.


BaMMV/BaYMV Hordeum vulgare L. marker-assisted selection Rym4/Rym5 SSRs winter barley 


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  1. Chalhoub BA, Thibault S, Laucou V, Rameau C, Höfte H, Cousin R, 1997. Silver staining and recovery of AFLP™ amplification products on large denaturing polyacrylamide gels. BioTechniques 22: 216–220.PubMedGoogle Scholar
  2. Graner A, Bauer E, 1993. RFLP mapping of theym4 virus resistance gene in barley. Theor Appl Genet 86: 689–693.CrossRefGoogle Scholar
  3. Graner A, Streng S, Kellermann A, Proeseler G, Schiemann A, Peterka H, Ordon F, 1999a. Molecular mapping of genes conferring resistance to soil-borne viruses in barley: an approach to promote understanding of host-pathogen interactions. J Plant Dis Prot 106: 405–410.Google Scholar
  4. Graner A, Streng S, Kellermann A, Schemann A, Bauer E, Waugh R, et al. 1999b. Molecular mapping and genetic fine-structure of therym5 locus encoding resistance to different strains of the Barley Yellow Mosaic Virus Complex. Theor Appl Genet 98: 285–290.CrossRefGoogle Scholar
  5. Harding RM, Boyce AJ, Clegg JB, 1992. The evolution of tandemly repetitive DNA: recombination rules. Genetics 132: 847–859.PubMedGoogle Scholar
  6. Kanyuka K, McGrann G, Alhudaib K, Hariri D, Adams MJ, 2004. Biological and sequence analysis of a novel European isolate ofBarley mild mosaic virus that overcomes the barleyrym5 resistance gene. Arch Virol 149: 1469–1480.CrossRefPubMedGoogle Scholar
  7. Konishi T, Ordon F, Furusho M, 2002. Reactions of barley accessions carrying different rym genes to BaYMV and BaMMV in Japan and Germany. Barley Genet Newsl 32: 46–48.Google Scholar
  8. Le Gouis J, Ordon F, Friedt W, 1994. Isozyme-electrophoresis for chromosomal localization of barley mild mosaic virus resistance genes different fromym4. Barley Genet Newsl 23: 36–39.Google Scholar
  9. Li Y-C, Korol AB, Fahima T, Beiles A, Nevo E, 2002. Microsatellites: genomic distribution, putative functions and mutational mechanisms: a review. Mol Ecol 11: 2453–2465.CrossRefPubMedGoogle Scholar
  10. McGrann GRD, Adams MJ, 2004. Investigating resistance toBarley mild mosaic virus. Plant Pathology 53: 161–169.CrossRefGoogle Scholar
  11. Milligan BG, 1992. Plant DNA isolation. In: Hoelzel AR, ed. Molecular analysis of populations: a practical approach. IRL Press, Oxford: 59–88.Google Scholar
  12. Morgante M, Hanafey M, Powell W, 2002. Microsatellites are preferentially associate with nonrepetitive DNA in plant genomes. Nature Genet 30: 194–200.CrossRefPubMedGoogle Scholar
  13. Ordon F, Bauer E, Dehmer KJ, Graner A, Friedt W, 1995a. Identification of a RAPD-marker linked to the BaMMV/BaYMV resistance geneym4. Barley Genet Newsl 24: 123–126.Google Scholar
  14. Ordon F, Bauer E, Graner A, Friedt W, 1995b. Marker-based selection for the BaMMV-resistance gene in barley using RAPDs. Agronomie 15: 481–485.CrossRefGoogle Scholar
  15. Ordon F, Friedt W, Scheurer K, Pellio B, Werner K, Neuhaus G, et al. 2004. Molecular markers in breeding for virus resistance in barley. J Appl Genet 45: 145–159.PubMedGoogle Scholar
  16. Pellio B, Friedt W, Graner A, Ordon F, 2004. Development of PCR-based markers closely linked torym5. J Plant Dis Prot 111: 30–38.Google Scholar
  17. Pellio B, Streng S, Bauer E, Stein N, Perovic D, Schiemann A, et al. 2005. High-resolution mapping of theRym4/Rym5 locus conferring resistance to the barley yellow mosaic virus complex (BaMMV, BaYMV, BaYMV-2) in barley (Hordeum vulgare ssp.vulgare L.). Theor Appl Genet 110: 283–293.CrossRefPubMedGoogle Scholar
  18. Röder MS, Huang XQ, Ganal MW, 2004. Wheat microsatellites: potential and implications. In: Lörz H, Wenzel G, eds. Biotechnology in agriculture and forestry: molecular marker systems. Springer Verlag: 255–266.Google Scholar
  19. Rozen S, Skaletsky H, 2000. Primer3 on the WWW for general users and for biologist programmers. In: Krawetz S, Misener S, eds. Bioinformatics Methods and Protocols: Methods in Molecular Biology, Humana Press, Totowa: 365–386.Google Scholar
  20. Ruge B, Linz A, Pickering R, Proeseler G, Greif P, Wehling P, 2003. Mapping of Rym14Hb, a gene introgressed fromHordeum bulbosum and conferring resistance to BaMMV and BaYMV in barley. Theor Appl Genet 107: 965–971.CrossRefPubMedGoogle Scholar
  21. Ruge-Wehling B, Linz A, Habekuß A, Wehling P, 2006. Mapping ofRym16 (Hb), the second soil-borne virus-resistance gene introgressed fromHordeum bulbosum. Theor Appl Genet 113: 867–873.CrossRefPubMedGoogle Scholar
  22. Schiemann A, Graner A, Friedt W, Ordon F, 1996. Specificity enhancement of a RAPD marker linked to the BaMMV/BaYMV resistance geneym4 by randomly added bases. Barley Genet Newsl 27: 63–65.Google Scholar
  23. Stein N, Perovic D, Kumlehn J, Pellio B, Stracke S, Streng S, et al. 2005. The eukaryotic translation initiation factor 4E confers multiallelic recessive Bymovirus resistance inHordeum vulgare (L.). Plant J 42: 912–922.CrossRefPubMedGoogle Scholar
  24. Tachida H, Iizuka M, 1992. Persistence of repeated sequences that evolve by replication slippage. Genetics 131: 471–478.PubMedGoogle Scholar
  25. Tatusova AT, Madden TL, 1999. Blast 2 sequences — a new tool for comparing protein and nucleotide sequences, FEMS Microbiol Lett 174: 247–250.CrossRefPubMedGoogle Scholar
  26. Thiel T, Kota R, Grosse I, Stein N, Graner A, 2004. SNP2CAPS: a SNP and INDEL analysis tool for CAPS marker development. Nucleic Acids Res 32: e5.CrossRefPubMedGoogle Scholar
  27. Thiel T, Michalek W, Varshney RK, Graner A, 2003. Exploiting EST databases for the development and characterization of gene-derived SSR-markers in barley (Hordeum vulgare L.). Theor Appl Genet 106: 41–422.Google Scholar
  28. Weber JL, 1990. Informativeness of human (dC.dA) n(dG.dT)n polymorphisms. Genomics 7: 524–530.CrossRefPubMedGoogle Scholar
  29. Werner K, Friedt W, Laubach E, Waugh R, Ordon F, 2003. Dissection of resistance to soil-borne yellow-mosaic-inducing viruses of barley (BaMMV, BaTMV, BaYMV-2) in a complex breeders’ cross by means of SSRs and simultaneous mapping of BaYMV/BaYMV-2 resistance of var. ‘Chikurin Ibaraki 1’. Theor Appl Genet 106: 1425–1432.PubMedGoogle Scholar
  30. Werner K, Friedt W, Ordon F, 2005. Strategies for pyramiding resistance genes against the barley yellow mosaic virus complex (BaMMV, BaYMV, BaYMV-2). Mol Breed 16: 45–55.CrossRefGoogle Scholar
  31. Wicker T, Zimmermann W, Perovic D, Paterson AH, Ganal M, Graner A, Stein N, 2005. A detailed look at 7 million years of genome evolution in a 439-kb contiguous sequence at the barley Hv-eIF4E locus: recombination, rearrangements and repeats. Plant J 41: 184–194.CrossRefPubMedGoogle Scholar

Copyright information

© Institute of Plant Genetics, Polish Academy of Sciences, Poznan 2008

Authors and Affiliations

  • Mirosław Tyrka
    • 1
  • Dragan Perovic
    • 2
  • Agnieszka Wardyńska
    • 1
  • Frank Ordon
    • 2
  1. 1.Laboratory of Population GeneticsPolonia UniversityCzęstochowaPoland
  2. 2.Federal Centre for Breeding Research on Cultivated PlantsInstitute of Epidemiology and Resistance ResourcesQuedlinburgGermany

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