Plant Molecular Biology

, Volume 65, Issue 1–2, pp 93–106 | Cite as

Leaf rust resistance gene Lr1, isolated from bread wheat (Triticum aestivum L.) is a member of the large psr567 gene family

  • Sylvie Cloutier
  • Brent D. McCallum
  • Caroline Loutre
  • Travis W. Banks
  • Thomas Wicker
  • Catherine Feuillet
  • Beat Keller
  • Mark C. Jordan
Article

Abstract

In hexaploid wheat, leaf rust resistance gene Lr1 is located at the distal end of the long arm of chromosome 5D. To clone this gene, an F1-derived doubled haploid population and a recombinant inbred line population from a cross between the susceptible cultivar AC Karma and the resistant line 87E03-S2B1 were phenotyped for resistance to Puccinia triticina race 1-1 BBB that carries the avirulence gene Avr1. A high-resolution genetic map of the Lr1 locus was constructed using microsatellite, resistance gene analog (RGA), BAC end (BE), and low pass (LP) markers. A physical map of the locus was constructed by screening a hexaploid wheat BAC library from cultivar Glenlea that is known to have Lr1. The locus comprised three RGAs from a gene family related to RFLP marker Xpsr567. Markers specific to each paralog were developed. Lr1 segregated with RGA567-5 while recombinants were observed for the other two RGAs. Transformation of the susceptible cultivar Fielder with RGA567-5 demonstrated that it corresponds to the Lr1 resistance gene. In addition, the candidate gene was also confirmed by virus-induced gene silencing. Twenty T1 lines from resistant transgenic line T0-938 segregated for resistance, partial resistance and susceptibility to Avr1 corresponding to a 1:2:1 ratio for a single hemizygous insertion. Transgene presence and expression correlated with the phenotype. The resistance phenotype expressed by Lr1 seemed therefore to be dependant on the zygosity status. T3-938 sister lines with and without the transgene were further tested with 16 virulent and avirulent rust isolates. Rust reactions were all as expected for Lr1 thereby providing additional evidence toward the Lr1 identity of RGA567-5. Sequence analysis of Lr1 indicated that it is not related to the previously isolated Lr10 and Lr21 genes and unlike these genes, it is part of a large gene family.

Keywords

Lr1 Leaf rust resistance gene Gene dosage VIGS Map-based cloning Triticum aestivum 

Supplementary material

References

  1. Altschul SF, Madden TL, Schaffer AA et al (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402PubMedCrossRefGoogle Scholar
  2. Ausemus ER, Harrington JB, Worzella WW et al (1946) A summary of genetic studies in hexaploid and tetraploid wheats. J Am Soc Agron 38:1082–1099Google Scholar
  3. Bai J, Pennill LA, Ning J et al (2002) Diversity in nucleotide binding site-leucine-rich repeat genes in cereals. Genome Res 12:1871–1884PubMedCrossRefGoogle Scholar
  4. Bossolini E, Wicker T, Knober PA et al (2007) Comparison of orthologous loci from small grass genomes Brachypodium and rice: implications from wheat genomics and grass genome annotation. Plant J 49:704–717PubMedCrossRefGoogle Scholar
  5. Brooks SA, Huang L, Herbel MN et al (2006) Structural variation and evolution of a defense-gene cluster in natural populations of Aegilops tauschii. Theor Appl Genet 112:618–626PubMedCrossRefGoogle Scholar
  6. Burch-Smith TM, Anderson JC, Martin GB et al (2004) Applications and advantages of virus-induced gene silencing for gene function studies in plants. Plant J 39:734–746PubMedCrossRefGoogle Scholar
  7. Chiu W, Niwa Y, Zeng W et al (1996) Engineered GFP as a vital reporter for plants. Curr Biol 6:325–330PubMedCrossRefGoogle Scholar
  8. Christensen AH, Quail PH (1996) Ubiquitin promoter-based vectors for high-level expression of selectable and/or screenable marker genes in monocotyledonous plants. Trans Res 5:213–218CrossRefGoogle Scholar
  9. Dangl JL, Jones JDG (2001) Plant pathogens and integrated defence responses to infection. Nature 411:826–833PubMedCrossRefGoogle Scholar
  10. Dyck PL, Samborski DJ (1968) Genetics of resistance to leaf rust in common wheat varieties. Webster, Loros, Brevit, Carina, Malakoff and Centenario. Can J Genet Cytol 10:7–17Google Scholar
  11. Dyck PL, Samborski DJ, Martens JW (1985) Inheritance of resistance to leaf rust and stem rust in the wheat cultivar Glenlea. Can J Plant Pathol 7:351–354Google Scholar
  12. Feuillet C, Messmer M, Schachermayr G et al (1995) Genetic and physical characterization of the LR1 leaf rust resistance locus in wheat (Triticum aestivum L.). Mol Gen Genet 248:553–562PubMedCrossRefGoogle Scholar
  13. Feuillet C, Travella S, Stein N et al (2003) Map-based isolation of the leaf rust disease resistance gene Lr10 from the hexaploid wheat (Triticum aestivum L.) genome. Proc Natl Acad Sci USA 100:15253–15258PubMedCrossRefGoogle Scholar
  14. Flor HH (1956) The complementary genic systems in flax and flax rust. Adv Genet 8:29–54CrossRefGoogle Scholar
  15. Gallego F, Feuillet C, Messmer M et al (1998) Comparative mapping of the two wheat leaf rust resistance loci Lr1 and Lr10 in rice and barley. Genome 41:328–336PubMedCrossRefGoogle Scholar
  16. Graham MA, Marek LF, Shoemaker RC (2002) Organization, expression and evolution of a disease resistance gene cluster in soybean. Genetics 162:1961–1977PubMedGoogle Scholar
  17. Graner A, Siedler H, Jahoor A et al (1990) Assessment of the degree and the type of restriction fragment length polymorphism in barley (Hordeum vulgare). Theor Appl Genet 80:826–832CrossRefGoogle Scholar
  18. Holzberg S, Brosio P, Gross C et al (2002) Barley stripe mosaic virus-induced gene silencing in a monocot plant. Plant J 30:315–327PubMedCrossRefGoogle Scholar
  19. Huang L, Brooks SA, Li W et al (2003) Map-based cloning of leaf rust resistance gene Lr21 from the large and polyploid genome of bread wheat. Genetics 164:655–664PubMedGoogle Scholar
  20. Hulbert SH, Webb CA, Smith SM et al (2001) Resistance gene complexes: evolution and utilization. Annu Rev Phytopathol 39:285–312PubMedCrossRefGoogle Scholar
  21. Iyer LM, Kumpatla SP, Chandrasekharan MB et al (2000) Transgene silencing in monocots. Plant Mol Biol 43:323–346PubMedCrossRefGoogle Scholar
  22. Jordan MC (2000) Green fluorescent protein as a visual marker for wheat transformation. Plant Cell Rep 19:1069–1075CrossRefGoogle Scholar
  23. Keller B, Feuillet C, Yahiaoui N (2005) Map-based isolation of disease resistance genes from bread wheat: cloning in a supersize genome. Genet Res 85:93–100PubMedCrossRefGoogle Scholar
  24. Kolmer JA (1996) Genetics of resistance to wheat leaf rust. Annu Rev Phytopathol 34:435–455PubMedCrossRefGoogle Scholar
  25. Kolmer JA (2001) Physiologic specialization of Puccinia triticina in Canada in 1998. Plant Dis 85:155–158CrossRefGoogle Scholar
  26. Kolmer JA (2005) Tracking wheat rust on a continental scale. Curr Opinion Plant Biol 8:441–449CrossRefGoogle Scholar
  27. Kolmer JA, Dyck PL (1994) Gene expression in the Triticum aestivum-Puccinia recondita f. sp. tritici gene-for-gene system. Phytopatol 84:437–440CrossRefGoogle Scholar
  28. Kolmer JA, Liu JQ (2002) Inheritance of leaf rust resistance in the wheat cultivars AC Majestic, AC Splendor, and AC Karma. Can J Plant Pathol 24:327–331CrossRefGoogle Scholar
  29. Kuang H, Woo S-S, Meyers BC et al (2004) Multiple genetic processes result in heterogeneous rates of evolution within the major cluster disease resistance genes in lettuce. Plant Cell 16:2870–2894PubMedCrossRefGoogle Scholar
  30. Lander ES, Green P, Abrahamson J et al (1987) MAPMAKER: an interactive computer package for constructing primary genetic linkage maps of experimental and natural populations. Genomics 1:174–181PubMedCrossRefGoogle Scholar
  31. Laurie DA, Bennett MD (1986) Wheat × maize hybridization. Can J Genet Cytol 28:313–316Google Scholar
  32. Lehmann P (2002) Structure and evolution of plant disease resistance genes. J Appl Genet 43:403–414PubMedGoogle Scholar
  33. Leister D (2004) Tandem and segmental gene duplication and recombination in the evolution of plant disease resistance genes. Trends Genet 20:116–122PubMedCrossRefGoogle Scholar
  34. Ling H-Q, Qiu J, Singh RP et al (2004) Identification and genetic characterization of an Aegilops tauschii ortholog of the wheat leaf rust disease resistance gene Lr1. Theor Appl Genet 109:1133–1138PubMedCrossRefGoogle Scholar
  35. Ling H-Q, Zhu Y, Keller B (2003) High-resolution mapping of the leaf rust disease resistance gene Lr1 in wheat and characterization of BAC clones from the Lr1 locus. Theor Appl Genet 106:875–882PubMedGoogle Scholar
  36. Liu JQ, Kolmer JA (1998) Genetics of stem rust resistance in wheat cvs Pasqua and AC Taber. Phytopathology 88:171–176CrossRefPubMedGoogle Scholar
  37. Long DL, Kolmer JA (1989) A North American system of nomenclature for Puccinia recondita f. sp. tritici. Phytopathology 79:525–529Google Scholar
  38. Luo MC, Thomas C, You FM et al (2003) High-throughput fingerprinting of bacterial artificial chromosomes using the snapshot labeling kit and sizing of restriction fragments by capillary electrophoresis. Genomics 82:378–389PubMedCrossRefGoogle Scholar
  39. Lupas A (1996) Coiled coils: new structures and new functions. Trends Biochem Sci 21:375–382PubMedCrossRefGoogle Scholar
  40. Lupas A, Van Dyke M, Stock J (1991) Predicting coiled coils from protein sequences. Science 252:1162–1164PubMedCrossRefGoogle Scholar
  41. Madsen LH, Collins NC, Rakwalska M et al (2003) Barley disease resistance gene analogs of the NBS-LRR class: identification and mapping. Mol Gen Genom 269:150–161Google Scholar
  42. Mains EB, Leighty CE, Johnston CO (1926) Inheritance of resistance to leaf rust Puccinia triticina Erikss., in crosses of common wheat, Triticum vulgare Vill. J Agric Res 32:931–972Google Scholar
  43. Martin GB, Bogdanove AJ, Sessa G (2003) Understanding the functions of plant disease resistance proteins. Annu Rev Plant Biol 54:23–61PubMedCrossRefGoogle Scholar
  44. McIntosh RA, Baker EP, Driscoll CS (1965) Cytogenetic studies in wheat I. Monosomic analysis of leaf rust resistance in cultivar Uruguay and Transfer. Aust J Biol Sci 18:971–977Google Scholar
  45. McVey DV (1989) Verification of infection-type data for identification of genes for resistance to leaf rust in some hard red spring wheats. Crop Sci 29:304–307CrossRefGoogle Scholar
  46. Meyers BC, Kaushik S, Nandety RS (2005) Evolving disease resistance genes. Curr Opin Plant Biol 8:129–134PubMedCrossRefGoogle Scholar
  47. Meyers BC, Kozik A, Griego A et al (2003) Genome-wide analysis of NBS-LRR-encoding genes in Arabidopsis. Plant Cell 15:809–834PubMedCrossRefGoogle Scholar
  48. Meyers BC, Shen KA, Rohani P et al (1998) Receptor-like genes in the major resistance locus of lettuce are subject to divergent selection. Plant Cell 11:1833–1846CrossRefGoogle Scholar
  49. Nilmalgoda SN, Cloutier S, Walichnowski AZ (2003) Construction and characterization of a bacterial artificial chromosome library of hexaploid wheat (Triticum aestivum L.) and validation of genome coverage using locus-specific primers. Genome 46:870–878PubMedCrossRefGoogle Scholar
  50. Radovanovic N, Cloutier S (2003) Gene-assisted selection for high molecular weight glutenin subunits in wheat doubled haploid breeding programs. Mol Breed 12:51–59CrossRefGoogle Scholar
  51. Samborski DJ (1973) Leaf rust of wheat in Canada in (1972). Can Plant Dis Surv 52:168–170Google Scholar
  52. Schachermayr GM, Messmer MM, Feuillet C et al (1995) Identification of molecular markers linked to the Agropyron elongatum-derived leaf rust resistance gene Lr24 in wheat. Theor Appl Genet 90:982–990CrossRefGoogle Scholar
  53. Scofield SR, Huang L, Brandt AS et al (2005) Development of a virus-induced gene-silencing system for hexaploid wheat and its use in functional analysis of the Lr21-mediated leaf rust resistance pathway. Plant Physiol 138:2165–2173PubMedCrossRefGoogle Scholar
  54. Shen Q-H, Sajo Y, Mauch S (2007) Nuclear activity of MLA immune receptors links isolate-specific and basal disease-resistance responses. Science 315:1098–1103PubMedCrossRefGoogle Scholar
  55. Simons K, Fellers JP, Trick HN et al (2005) Isolation and characterization of the major domestication gene Q in wheat. Proc Plant Ani Genome XIII meeting, 15–19 Jan 2005, San Diego, USA, p 97Google Scholar
  56. Somers DJ, Isaac P, Edwards K (2004) A high-density microsatellite consensus map for bread wheat (Triticum aestivum L.). Theor Appl Genet 109:1105–1114PubMedCrossRefGoogle Scholar
  57. Sonnhammer EL, Durbin R (1995) A dot-matrix program with dynamic threshold control suited for genomic DNA and protein sequence analysis. Gene 167:1–10CrossRefGoogle Scholar
  58. Stam M, Mol JNM, Kooter JM (1997) The silence of genes in transgenic plants. Ann Bot 79:3–12CrossRefGoogle Scholar
  59. Stein N, Feuillet C, Wicker T et al (2000) Subgenome chromosome walking in wheat: a 450-kb physical contig in Triticum monococcum L. spans the Lr10 resistance locus in hexaploid wheat (Triticum aestivum L.). Proc Natl Acad Sci 97:13436–13441PubMedCrossRefGoogle Scholar
  60. Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680PubMedCrossRefGoogle Scholar
  61. Upadhyaya NM, Ramm K, Gaudron J et al (1998) Can gfp replace uidA as a reporter gene to monitor transformation of cereals by biolistics or Agrobacterium? In: Larkin PJ (ed) Agricultural Biotechnology: Laboratory, Field and Market. Proceedings of the 4th Asia-Pacific conference on agricultural biotechnology, Darwin, 13–16 July, Canberra UTC publishing, pp 111–113Google Scholar
  62. Van der Biezen EA, Jones JDG (1998) Plant disease-resistance proteins and the gene-for-gene concept. Trends Biochem Sci 23:454–456PubMedCrossRefGoogle Scholar
  63. Vaucheret H, Béclin C, Elmayan T et al (1998) Transgene-induced gene silencing in plants. Plant J 16:651–659PubMedCrossRefGoogle Scholar
  64. Webb CA, Fellers JP (2006) Cereal rust fungi genomics and the pursuit of virulence and avirulence factors. FEMS Microbiol Lett 264:1–7PubMedCrossRefGoogle Scholar
  65. Xie DX, Devos KM, Moore G et al (1993) RFLP-based genetic maps of the homoeologous group 5 chromosomes of bread wheat (Triticum aestivum L.). Theor Appl Genet 87:70–74CrossRefGoogle Scholar
  66. Xu Z, Deal KR, Li W et al (2002) Construction and characterization of five large-insert BAC and BiBAC libraries of Aegilops tauschii, the diploid donor of the wheat D genome. Proc Plant Ani Microbe Genomes X meeting, 12–16 Jan 2002, San Diego, USA, p 101Google Scholar
  67. Yahiaoui N, Srichumpa P, Dudler R et al (2004) Genome analysis at different ploidy levels allows cloning of the powdery mildew resistance gene Pm3b from hexaploid wheat. Plant J 37:528–538PubMedCrossRefGoogle Scholar
  68. Yan L, Loukoianov A, Tranquilli G et al (2003) Positional cloning of the wheat vernalization gene VRN1. Proc Natl Acad Sci USA 100:6263–6268PubMedCrossRefGoogle Scholar
  69. Yan L, Loukoianov A, Blechl A et al (2004) The wheat VRN2 gene is a flowering repressor down-regulated by vernalization. Science 303:1640–1644PubMedCrossRefGoogle Scholar
  70. Zhang H-B, Zhao Z, Ding X et al (1995) Preparation of megabase-size DNA from plant nuclei. Plant J 7:175–184CrossRefGoogle Scholar
  71. Zhou T, Wang Y, Chen JQ et al (2004) Genome-wide identification of NBS genes in japonica rice reveals significant expansion of divergent non-TIR NBS-LRR genes. Mol Genet Genom 271:402–415CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2007

Authors and Affiliations

  • Sylvie Cloutier
    • 1
  • Brent D. McCallum
    • 1
  • Caroline Loutre
    • 2
  • Travis W. Banks
    • 1
  • Thomas Wicker
    • 2
  • Catherine Feuillet
    • 2
    • 3
  • Beat Keller
    • 2
  • Mark C. Jordan
    • 1
  1. 1.Cereal Research CentreAgriculture and Agri-Food CanadaWinnipegCanada
  2. 2.Institute of Plant BiologyUniversity of ZürichZurichSwitzerland
  3. 3.Institut National de la Recherche AgronomiqueAmélioration et Santé des PlantesClermont FerrandFrance

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