Theoretical and Applied Genetics

, Volume 126, Issue 1, pp 93–99 | Cite as

Molecular tagging of a novel rust resistance gene R 12 in sunflower (Helianthus annuus L.)

  • L. Gong
  • B. S. Hulke
  • T. J. Gulya
  • S. G. Markell
  • L. L. Qi
Original Paper

Abstract

Sunflower production in North America has recently suffered economic losses in yield and seed quality from sunflower rust (Puccinia helianthi Schwein.) because of the increasing incidence and lack of resistance to new rust races. RHA 464, a newly released sunflower male fertility restorer line, is resistant to both of the most predominant and most virulent rust races identified in the Northern Great Plains of the USA. The gene conditioning rust resistance in RHA 464 originated from wild Helianthus annuus L., but has not been molecularly marked or determined to be independent from other rust loci. The objectives of this study are to identify molecular markers linked to the rust resistance gene and to investigate the allelism of this gene with the unmapped rust resistance genes present in HA-R6, HA-R8 and RHA 397. Virulence phenotypes of seedlings for the F2 population and F2:3 families suggested that a single dominant gene confers rust resistance in RHA 464, and this gene was designated as R 12 . Bulked segregant analysis identified ten markers polymorphic between resistant and susceptible bulks. In subsequent genetic mapping, the ten markers covered 33.4 cM of genetic distance on linkage group 11 of sunflower. A co-dominant marker CRT275-11 is the closest marker distal to R 12 with a genetic distance of 1.0 cM, while ZVG53, a dominant marker linked in the repulsion phase, is proximal to R 12 with a genetic distance of 9.6 cM. The allelism test demonstrated that R 12 is not allelic to the rust resistance genes in HA-R6, HA-R8 and RHA 397, and it is also not linked to any previously mapped rust resistance genes. Discovery of the R 12 novel rust resistance locus in sunflower and associated markers will potentially support the molecular marker-assisted introgression and pyramiding of R 12 into sunflower breeding lines.

Keywords

Rust Resistance Downy Mildew Infection Type Rust Resistance Gene Helianthus Annuus 

Notes

Acknowledgments

We thank Drs. Gerald Seiler and Steven Xu for critical review of the manuscript and Angelia Hogness for technical assistance. This project was supported by the USDA-AMS Cooperative Agreement 12-25-B-0942.

References

  1. Bachlava E, Radwan OE, Abratti G, Tang S, Gao W, Heesacker AF, Bazzalo ME, Zambelli A, Leon AJ, Knapp SJ (2011) Downy mildew (Pl 8 and Pl 14) and rust (R Adv) resistance genes reside in close proximity to tandemly duplicated clusters of non-TIR-like NBS-LRR-encoding genes on sunflower chromosomes 1 and 13. Theor Appl Genet 122:1211–1221PubMedCrossRefGoogle Scholar
  2. Bachlava E, Taylor CA, Tang S, Bowers JE, Mandel JR, Burke JM, Knapp SJ (2012) SNP discovery and development of a high-density genotyping array for sunflower. PLoS ONE 7(1):e29814. doi: 10.1371/journal.pone.0029814 PubMedCrossRefGoogle Scholar
  3. Bert PF, De Labrouhe DT, Philippon J, Mouzeyar S, Jouan I, Nicolas P, Vear F (2001) Identification of a second linkage group carrying genes controlling resistance to downy mildew (Plasmopara halstedii) in sunflower (Helianthus annuus L.). Theor Appl Genet 103:992–997CrossRefGoogle Scholar
  4. Bouzidi MF, Badaoui S, Cambon F, Vear F, De Labrouhe DT, Nicolas P, Mouzeyar S (2002) Molecular analysis of a major locus for resistance to downy mildew in sunflower with specific PCR-based markers. Theor Appl Genet 104:592–600PubMedCrossRefGoogle Scholar
  5. Bowers JE, Bachlava E, Brunick RL, Rieseberg LH, Knapp SJ, Burke JM (2012) Development of a 10,000 locus genetic map of the sunflower genome based on multiple crosses. Genes Genomes Genetics 2:721–729PubMedGoogle Scholar
  6. Burke JM, Tang S, Knapp SJ, Rieseberg LH (2002) Genetic analysis of sunflower domestication. Genetics 161:1257–1267PubMedGoogle Scholar
  7. de Romano AB, Romano C, Bulos M, Altieri E, Sala C (2010) A new gene for resistance to downy mildew in sunflower. In: Proc Int Symposium “Sunflower breeding on resistance to diseases”, Krasnodar, Russia, June 23–24Google Scholar
  8. Del Moral L, Fernández-Martínez JM, Velasco L, Pérez-Vich B (2012) Quantitative trait loci for seed tocopherol content in sunflower. Crop Sci 52:786–794Google Scholar
  9. Dußle CM, Hahn V, Knapp SJ, Bauer E (2004) PI Arg from Helianthus argophyllus is unlinked to other known downy mildew resistance genes in sunflower. Theor Appl Genet 109:1083–1086CrossRefGoogle Scholar
  10. Friskop A, Gulya T, Jordahl J, Ramsett M, Harveson R, Acevedo M, Harveson R, Markell S (2012) Determination of Puccinia helianthi races in the Unites States Northern Great Plains. In: Proceedings of the 18th international sunflower conference, Mardel plata and Balcarce, Argentina Feb 27–Mar 1, 2012. pp 214–218Google Scholar
  11. Gedil MA, Slabaugh MB, Berry S, Segers B, Peleman J, Michelmore R, Miller JF, Gulya T, Knapp SJ (2001) Candidate disease resistance genes in sunflower cloned using conserved nucleotide binding site motifs: genetic mapping and linkage to downy mildew resistance gene Pl 1. Genome 44:205–212PubMedGoogle Scholar
  12. Gentzbittel L, Mouzeyar F, Badaoui S, Mestries E, Vear F, de Labrouhe DT, Nicolas P (1998) Cloning of molecular markers for disease resistance in sunflower, Helianthus annuus L. Theor Appl Genet 96:519–525CrossRefGoogle Scholar
  13. Goulter KC (1990) Breeding of a rust differential sunflower line. In: Proceedings of the 8th Australian sunflower association workshop, Toowoomba, 19–22 Mar 1990, pp 120–124Google Scholar
  14. Gulya TJ (2006) The sunflower rust situation: current races in the northern and central Great Plains, and resistance in oilseed and confection hybrids. In: Proceedings of 28th sunflower research workshop. Fargo, ND. 11–12 Jan 2006. http://www.sunflowernsa.com/research/research-workshop/documents/Gulya_Rust_06.pdf
  15. Gulya TJ, Markell S (2009) Sunflower rust status—2008. Race frequency across the midwest and resistance among commercial hybrids. http://www.sunflowernsa.com/uploads/Gulya_RustStatus_09.pdf
  16. Gulya TJ, Masirevic S (1996) Inoculation and evaluation methods for sunflower rust. In: Proceedings of 18th sunflower research workshop. Fargo, ND, 11–12 Jan 1996, pp 31–38Google Scholar
  17. Gulya TJ, Venette R, Venette JR, Lamey HA (1990) Sunflower rust. NDSU Ext. Ser. Bull., Fargo, p 998. http://www.ag.ndsu.edu/pubs/plantsci/rowcrops/pp998w.htm
  18. Gulya TJ, Markell S, McMullen M, Harveson B, Osborne L (2011) New virulent races of downy mildew: distribution, status of DM resistant hybrids, and USDA sources of resistance. In: Proc 33th Sunflower Research Forum. Fargo ND, January 12–13 http://www.sunflowernsa.com/uploads/resources/575/gulya_virulentracesdownymildew.pdf
  19. Heesacker A, Kishore VK, Gao W, Tang S, Kolkman JM, Gingle A, Matvienko M, Kozik A, Michelmore RM, Lai Z, Rieseberg LH, Knapp SJ (2008) SSRs and INDELs mined from the sunflower EST database: abundance, polymorphisms, and cross-taxa utility. Theor Appl Genet 117:1021–1029PubMedCrossRefGoogle Scholar
  20. Hulke BS, Miller JF, Gulya TJ (2010) Registration of the restorer oilseed sunflower germplasm RHA 464 possessing genes for resistance to downy mildew and sunflower rust. J Plant Regist 4:249–254CrossRefGoogle Scholar
  21. Jan CC, Gulya TJ (2006) Registration of a sunflower germplasm resistant to rust, downy mildew and virus. Crop Sci 46:1829CrossRefGoogle Scholar
  22. Jan CC, Quresh Z, Gulya TJ (2004) Registration of seven rust resistance sunflower germplasms. Crop Sci 44:1887–1888CrossRefGoogle Scholar
  23. Kane NC, Gill N, King MG, Bowers JE, Berges H, Gouzy J, Bachlava E, Langlade NB, Lai Z, Stewart M, Burke JM, Vincourt P, Knapp SJ, Rieseberg LH (2011) Progress towards a reference genome for sunflower. Botany 89:429–437CrossRefGoogle Scholar
  24. Lambrides CJ, Miller JF (1994) Inheritance of rust resistance in a source of MC29 sunflower germplasm. Crop Sci 34:1225–1230CrossRefGoogle Scholar
  25. Lawson WR, Goulter KC, Henry RJ, Kong GA, Kochman JK (1998) Marker-assisted selection for two rust resistance genes in sunflower. Mol Breeding 4:227–234CrossRefGoogle Scholar
  26. Lawson WR, Jan CC, Shatte T, Smith L, Kong GA, Kochman JK (2011) DNA markers linked to the R 2 rust resistance gene in sunflower (Helianthus annuus L.) facilitate anticipatory breeding for this disease variant. Mol Breed 28:569–576CrossRefGoogle Scholar
  27. Liu Z, Gulya T, Seiler G, Vick BA, Jan CC (2012) Molecular mapping of the Pl 16 downy mildew resistance gene from HA-R4 to facilitate marker-assisted selection in sunflower. Theor Appl Genet. doi: 10.1007/s00122-012-1820-z Google Scholar
  28. Markell S, Gulya T, McKay K, Hutter M, Hollingsworth C, Ulstad V, Koch R, Knudsvig A (2009) Widespread occurrence of the aecial stage of sunflower rust caused by Puccinia helianthi in North Dakota and Minnesota in 2008. Plant Dis 93:668–669CrossRefGoogle Scholar
  29. Miah MAJ, Sackston WE (1970) Genetics of host-pathogen interaction in sunflower. Phytoprotection 51:1–16Google Scholar
  30. Michelmore RW, Paran I, Kesseli RV (1991) Identification of markers linked to disease-resistance genes by bulked segregant analysis: a rapid method to detect markers in specific genomic regions by using segregating populations. Proc Natl Acad Sci USA 88:9828–9832PubMedCrossRefGoogle Scholar
  31. Miller JF, Gulya TJ (1997) Registration of eight maintainer (HA 393, HA 394 and HA 402 to HA 407) and seven restorer (RHA 395 to RHA 401) sunflower germplasm lines. Crop Sci 37:1988–1989CrossRefGoogle Scholar
  32. Miller JF, Gulya TJ (2001) Registration of three rust resistant sunflower germplasm populations. Crop Sci 41:601CrossRefGoogle Scholar
  33. Miller JF, Rodriguez RH, Gulya TJ (1988) Evaluation of genetic materials for inheritance of resistance to Race 4 rust in sunflower. In: Proceedings of the 12th international sunflower conference, Novi Sad, Yugoslavia, 25–29 July 1988. International Sunflower Association, Paris, pp 361–365Google Scholar
  34. Moreno PS, de Romano AB, Romano MC, Vergani P, Sposaro M, Bulos M, Altieri E, Ramos ML, Sala C (2012) A survey of physiological races of Puccinia helianthi in Argentina. In: Proceedings of the 18th international sunflower conference, Mardel plata & Balcarce, Argentina Feb 27–Mar 1, 2012. pp 278–279Google Scholar
  35. Mouzeyar S, Roeckel-Drevet P, Gentzbittel L, Philippon J, de Labrouhe DT, Vear F, Nicolas P (1995) RFLP and RAPD mapping of the sunflower Pl 1 locus for resistance to Plasmopara halstedii race 1. Theor Appl Genet 91:733–737CrossRefGoogle Scholar
  36. Mulpuri S, Liu Z, Feng J, Gulya TJ, Jan CC (2009) Inheritance and molecular mapping of a downy mildew resistance gene, Pl 13 in cultivated sunflower (Helianthus annuus L.). Theor Appl Genet 119:795–803PubMedCrossRefGoogle Scholar
  37. Panković D, Radovanović N, Jocić S, Satovic Z, Skorić D (2007) Development of co-dominant amplified polymorphic sequence markers for resistance of sunflower to downy mildew race 730. Plant Breed 126:440–444CrossRefGoogle Scholar
  38. Putt ED, Sackston WE (1963) Studies on sunflower rust. IV. Two genes, R 1 and R 2 for resistance in the host. Can J Plant Sci 43:490–496CrossRefGoogle Scholar
  39. Qi LL, Gulya TJ, Seiler GJ, Hulke BS, Vick BA (2011a) Identification of resistance to new virulent races of rust in sunflowers and validation of DNA markers in the gene pool. Phytopathology 101:241–249PubMedCrossRefGoogle Scholar
  40. Qi LL, Hulke BS, Vick BA, Gulya TJ (2011b) Molecular mapping of the rust resistance gene R 4 to a large NBS-LRR cluster on linkage group 13 of sunflower. Theor Appl Genet 123:351–358PubMedCrossRefGoogle Scholar
  41. Qi LL, Gulya TJ, Hulke BS, Vick BA (2012a) Chromosome location, DNA markers and rust resistance of the sunflower gene R 5. Mol Breed 30:745–756CrossRefGoogle Scholar
  42. Qi LL, Seiler GJ, Hulke BS, Vick BA, Gulya TJ (2012b) Genetics and mapping of the R 11 gene conferring resistance to recently emerged rust races, tightly linked to male fertility restoration, in sunflower (Helianthus annuus L.) Theor Appl Genet. doi:  10.1007/s00122-012-1883-x
  43. Radwan O, Bouzidi MF, Vear F, Philippon J, de Labrouhe DT, Nicolas P, Mouzeyar S (2003) Identification of non-TIR-NBS-LRR markers linked to the Pl 5 /Pl 8 locus for resistance to downy mildew in sunflower. Theor Appl Genet 106:1438–1446PubMedGoogle Scholar
  44. Radwan O, Bouzidi MF, Nicolas P, Mouzeyar S (2004) Development of PCR markers for the Pl 5 /Pl 8 locus for resistance to Plasmopara halstedii in sunflower, Helianthus annuus L. from complete CC-NBS-LRR sequences. Theor Appl Genet 109:176–185PubMedCrossRefGoogle Scholar
  45. Roeckel-Drevet P, Gagne G, Mouzeyar S, Gentzbittel L, Philippon J, Nicolas P, de Labrouhe DT, Vear F (1996) Colocation of downy mildew (Plasmopara halstedii) resistance genes in sunflower (Helianthus annuus L.). Euphytica 91:225–228Google Scholar
  46. Seiler GJ, Jan CC (1997) Registration of 10 interspecific germplasm fertility restoration populations for sunflower PET1 male-sterile cytoplasm. Crop Sci 37:1989–1991CrossRefGoogle Scholar
  47. Stam P (1993) Construction of integrated genetic linkage maps by means of a new computer package: JoinMap. Plant J 3:739–744CrossRefGoogle Scholar
  48. Tang S, Yu JK, Slabaugh MB, Shintani DK, Knapp SJ (2002) Simple sequence repeat map of the sunflower genome. Theor Appl Genet 105:1124–1136PubMedCrossRefGoogle Scholar
  49. Tang S, Kishore VK, Knapp SJ (2003) PCR-multiplexes for a genome-wide framework of simple sequence repeat marker loci in cultivated sunflower. Theor Appl Genet 107:6–19PubMedGoogle Scholar
  50. Vear F, Gentzbittel L, Philippon J, Mouzeyar S, Mestries E, Roeckel-Drevet P, de Labroube DT, Nicolas P (1997) The genetics of resistance to five races of downy mildew (Plasmopara halstedii) in sunflower (Helianthus annuus L.). Theor Appl Genet 95:584–589CrossRefGoogle Scholar
  51. Venkatramana P, Hulke B, Qi LL, Schultz Q (2012) SNP assisted breeding for sunflower development. In: Proceedings of the 18th international sunflower conference, Argentina, Feb 27–Mar 2012. International Sunflower Association, Paris, pp 1018–1019Google Scholar
  52. Wieckhorst S, Bachlava E, Dußle CM, Tang S, Gao W, Saski C, Knapp SJ, Schön CC, Hahn V, Bauer E (2010) Fine mapping of the sunflower resistance locus Pl ARG introduced from the wild species Helianthus argophyllus. Theor Appl Genet 121:1633–1644PubMedCrossRefGoogle Scholar
  53. Yang SM, Antonelli EF, Luciano A, Luciani ND (1986) Reactions of Argentine and Australian sunflower rust differentials to four North American cultures of Puccinia helianthi from North Dakota. Plant Dis 70:883–886CrossRefGoogle Scholar
  54. Yang SM, Dowler WM, Luciano A (1989) Gene Pu 6: a new gene in sunflower for resistance to Puccinia helianthi. Phytopathology 79:474–477CrossRefGoogle Scholar
  55. Yu JK, Tang S, Slabaugh MB, Heesacker A, Cole G, Herring M et al (2003) Towards a saturated molecular genetic linkage map for cultivated sunflower. Crop Sci 43:367–387CrossRefGoogle Scholar

Copyright information

© Springer-Verlag (outside the USA) 2012

Authors and Affiliations

  • L. Gong
    • 1
  • B. S. Hulke
    • 2
  • T. J. Gulya
    • 2
  • S. G. Markell
    • 1
  • L. L. Qi
    • 2
  1. 1.Department of Plant PathologyNorth Dakota State UniversityFargoUSA
  2. 2.Northern Crop Science LaboratoryUSDA, Agricultural Research ServiceFargoUSA

Personalised recommendations