Register      Login
Crop and Pasture Science Crop and Pasture Science Society
Plant sciences, sustainable farming systems and food quality
RESEARCH ARTICLE

Molecular mapping and validation of Rlm1 gene for resistance to Leptosphaeria maculans in canola (Brassica napus L.)

Rosy Raman A D , Belinda Taylor A , Kurt Lindbeck A , Neil Coombes A , Denise Barbulescu B , Phil Salisbury B C and Harsh Raman A D E
+ Author Affiliations
- Author Affiliations

A EH Graham Centre for Agricultural Innovation (an alliance between NSW Department of Primary Industries and Charles Sturt University), Wagga Wagga Agricultural Institute, PMB, Wagga Wagga, NSW 2650, Australia.

B Department of Primary Industries, Private Bag 260, Horsham, Vic. 3401, Australia.

C Department of Agriculture and Food Systems, Melbourne School of Land and Environment, The University of Melbourne, Vic. 3010, Australia.

D NSW Department of Primary Industries, Wagga Wagga Agricultural Institute, PMB, Wagga Wagga, NSW 2650, Australia.

E Corresponding author. Email: harsh.raman@dpi.nsw.gov.au

Crop and Pasture Science 63(10) 1007-1017 https://doi.org/10.1071/CP12255
Submitted: 12 July 2012  Accepted: 23 October 2012   Published: 18 December 2012

Abstract

European winter canola (Brassica napus L.) cultivars harbour genes for durable resistance to the fungus Leptosphaeria maculans, which causes blackleg disease under Australian environmental conditions. Previous studies have shown that resistance in winter-type cultivars Maxol and Columbus is controlled by two genes, Rlm1 and Rlm3, which have been mapped using randomly amplified polymorphic DNA markers onto chromosome A7. We mapped a doubled-haploid population that consisted of 101 lines from a cross between Maxol*1 and Westar-10 using diversity arrays technology and simple sequence repeat (SSR)-based markers. Two SSR marker loci, Xol12-e03 and Xra2-a05b, flanked the Rlm1 locus at an interval of 6.7 cM, which corresponds to ~3.2 Mb of the Brassica rapa genomic sequence; this region contains several genes encoding putative kinase and leucine-rich repeat-type disease-resistance proteins. SSR markers were further tested for their linkage with the Rlm1 locus in an independent population derived from Columbus*3/Westar-10. Our results showed that SSR markers linked to Rlm1 can be useful for monitoring Rlm1 gene introgression in breeding populations derived from Maxol and Columbus.

Additional keywords: blackleg, marker-assisted selection, Rlm1, Rlm3, race specific resistance genes, simple sequence repeat markers.


References

Aubertot J, West J, Bousset-Vaslin L, Salam M, Barbetti M, Diggle A (2006) Improved resistance management for durable disease control: A case study of phoma stem canker of oilseed rape (Brassica napus). European Journal of Plant Pathology 114, 91–106.
Improved resistance management for durable disease control: A case study of phoma stem canker of oilseed rape (Brassica napus).Crossref | GoogleScholarGoogle Scholar |

Balesdent MH, Attard A, Ansan-Melayah D, Delourme R, Renard M, Rouxel T (2001) Genetic control and host range of avirulence toward Brassica napus cultivars Quinta and Jet Neuf in Leptosphaeria maculans. Phytopathology 91, 70–76.
Genetic control and host range of avirulence toward Brassica napus cultivars Quinta and Jet Neuf in Leptosphaeria maculans.Crossref | GoogleScholarGoogle Scholar |

Balesdent MH, Attard A, Kuhn ML, Rouxel T (2002) New avirulence genes in the phytopathogenic fungus Leptosphaeria maculans. Phytopathology 92, 1122–1133.
New avirulence genes in the phytopathogenic fungus Leptosphaeria maculans.Crossref | GoogleScholarGoogle Scholar |

Balesdent MH, Louvard K, Pinochet X, Rouxel T (2006) A large-scale survey of races of Leptosphaeria maculans occurring on oilseed rape in France. European Journal of Plant Pathology 114, 53–65.
A large-scale survey of races of Leptosphaeria maculans occurring on oilseed rape in France.Crossref | GoogleScholarGoogle Scholar |

Barbulescu DM, Burton WA, Salisbury PA (2011) Pluronic F-68: an answer for shoot regeneration recalcitrance in microspore-derived Brassica napus embryos. In Vitro Cellular & Developmental Biology. Plant 47, 282–288.
Pluronic F-68: an answer for shoot regeneration recalcitrance in microspore-derived Brassica napus embryos.Crossref | GoogleScholarGoogle Scholar |

Bohman S, Wang M, Dixelius C (2002) Arabidopsis thaliana-derived resistance against Leptosphaeria maculans in a Brassica napus genomic background. Theoretical and Applied Genetics 105, 498–504.
Arabidopsis thaliana-derived resistance against Leptosphaeria maculans in a Brassica napus genomic background.Crossref | GoogleScholarGoogle Scholar |

Brun H, Fitt BDL, Powers S, Besnard A-L, Ermel M, Huteau V, Marquer B, Renard M, Andrivon D (2010) Quantitative resistance increases the durability of qualitative resistance to Leptosphaeria maculans in Brassica napus. New Phytologist 185, 285–299.
Quantitative resistance increases the durability of qualitative resistance to Leptosphaeria maculans in Brassica napus.Crossref | GoogleScholarGoogle Scholar |

Butler DG, Cullis BR, Gilmour AR, Gogel BJ (2009) ASReml-R reference manual. Release 3.0. Technical Report, Queensland Department of Primary Industries, Australia.

Chèvre AM, Barret P, Eber F, Dupuy P, Brun H, Tanguy X, Renard M (1997) Selection of stable Brassica napus–B. juncea recombinant lines resistant to blackleg (Leptosphaeria maculans). 1. Identification of molecular markers, chromosomal and genomic origin of the introgression. Theoretical and Applied Genetics 95, 1104–1111.

Christianson JA, Rimmer SR, Good AG, Lydiate DJ (2006) Mapping genes for resistance to Leptosphaeria maculans in Brassica juncea. Genome 49, 30–41.
Mapping genes for resistance to Leptosphaeria maculans in Brassica juncea.Crossref | GoogleScholarGoogle Scholar |

Coombes NE (2002) The reactive tabu search for efficient correlated experimental designs. PhD Thesis, John Moores University, Liverpool, UK.

Dangl JL, Jones JDG (2001) Plant pathogens and integrated defence responses to infection. Nature 411, 826–833.
Plant pathogens and integrated defence responses to infection.Crossref | GoogleScholarGoogle Scholar |

Delourme R, Pilet-Nayel ML, Archipiano M, Horvais R, Tanguy X, Rouxel T, Brun H, Renard M, Balesdent MH (2004) A cluster of major specific resistance genes to Leptosphaeria maculans in Brassica napus. Phytopathology 94, 578–583.
A cluster of major specific resistance genes to Leptosphaeria maculans in Brassica napus.Crossref | GoogleScholarGoogle Scholar |

Delourme R, Chevre AM, Brun H, Rouxel T, Balesdent MH, Dias JS, Salisbury P, Renard M, Rimmer SR (2006) Major gene and polygenic resistance to Leptosphaeria maculans in oilseed rape (Brassica napus). European Journal of Plant Pathology 114, 41–52.
Major gene and polygenic resistance to Leptosphaeria maculans in oilseed rape (Brassica napus).Crossref | GoogleScholarGoogle Scholar |

Dion Y, Gugel RK, Rakow GFW, Séguin-Swartz G, Landry BS (1995) RFLP mapping of resistance to the blackleg disease [causal agent, Leptosphaeria maculans (Desm.) Ces. et de Not.] in canola (Brassica napus L.). Theoretical and Applied Genetics 91, 1190–1194.
RFLP mapping of resistance to the blackleg disease [causal agent, Leptosphaeria maculans (Desm.) Ces. et de Not.] in canola (Brassica napus L.).Crossref | GoogleScholarGoogle Scholar |

Ellis J, Jones D (1998) Structure and function of proteins controlling strain-specific pathogen resistance in plants. Current Opinion in Plant Biology 1, 288–293.
Structure and function of proteins controlling strain-specific pathogen resistance in plants.Crossref | GoogleScholarGoogle Scholar |

Ferreira ME, Rimmer SR, Williams PH, Osborn TC (1995) Mapping loci controlling Brassica napus resistance to Leptosphaeria maculans under different screening conditions Phytopathology 85, 213–217.
Mapping loci controlling Brassica napus resistance to Leptosphaeria maculans under different screening conditionsCrossref | GoogleScholarGoogle Scholar |

Fitt B, Evans N, Howlett B, Cooke M (2006) Sustainable strategies for managing Brassica napus (Oilseed Rape) resistance to Leptosphaeria maculans (Phoma Stem Canker). European Journal of Plant Pathology 114, 1
Sustainable strategies for managing Brassica napus (Oilseed Rape) resistance to Leptosphaeria maculans (Phoma Stem Canker).Crossref | GoogleScholarGoogle Scholar |

Leflon M, Brun H, Eber F, Delourme R, Lucas MO, Vallee P, Ermel M, Balesdent MH, Chevre AM (2007) Detection, introgression and localization of genes conferring specific resistance to Leptosphaeria maculans from Brassica rapa into B. napus. Theoretical and Applied Genetics 115, 897–906.
Detection, introgression and localization of genes conferring specific resistance to Leptosphaeria maculans from Brassica rapa into B. napus.Crossref | GoogleScholarGoogle Scholar |

Light KA, Gororo NN, Salisbury PA (2011) Usefulness of winter canola (Brassica napus) race-specific resistance genes against blackleg (causal agent Leptosphaeria maculans) in southern Australian growing conditions. Crop & Pasture Science 62, 162–168.
Usefulness of winter canola (Brassica napus) race-specific resistance genes against blackleg (causal agent Leptosphaeria maculans) in southern Australian growing conditions.Crossref | GoogleScholarGoogle Scholar |

Long Y, Wang Z, Sun Z, Fernando DW, McVetty PB, Li G (2011) Identification of two blackleg resistance genes and fine mapping of one of these two genes in a Brassica napus canola cultivar ‘Surpass 400’. Theoretical and Applied Genetics 122, 1223–1231.
Identification of two blackleg resistance genes and fine mapping of one of these two genes in a Brassica napus canola cultivar ‘Surpass 400’.Crossref | GoogleScholarGoogle Scholar |

Lowe AJ, Moule C, Trick M, Edwards KJ (2004) Efficient large-scale development of microsatellites for marker and mapping applications in Brassica crop species. Theoretical and Applied Genetics 108, 1103–1112.
Efficient large-scale development of microsatellites for marker and mapping applications in Brassica crop species.Crossref | GoogleScholarGoogle Scholar |

Manly KF, Cudmore JRH, Meer JM (2001) MapManager QTX, cross platform software for genetic mapping. Mammalian Genome 12, 930–932.
MapManager QTX, cross platform software for genetic mapping.Crossref | GoogleScholarGoogle Scholar |

Marcroft S, Bluett C (2008) Blackleg of canola. Agriculture Notes, State of Victoria, Department of Primary Industries, May 2008, AG1352.

Marcroft SJ, Purwantara A, Salisbury P, Potter TD, Wratter N, Khangura R, Barbetti MJ, Howlett BJ (2002) Reaction of a range of Brassica species under Australian conditions to the fungus, Leptosphaeria maculans, the causal agent of blackleg. Australian Journal of Experimental Agriculture 42, 587–594.
Reaction of a range of Brassica species under Australian conditions to the fungus, Leptosphaeria maculans, the causal agent of blackleg.Crossref | GoogleScholarGoogle Scholar |

Marcroft S, van De Wouw A, Salisbury P, Potter T, Howlett BJ (2012a) Rotation of canola (Brassica napus) cultivars with differential complements of blackleg resistance genes decreases disease severity. Plant Pathology
Rotation of canola (Brassica napus) cultivars with differential complements of blackleg resistance genes decreases disease severity.Crossref | GoogleScholarGoogle Scholar |

Marcroft SJ, Elliott VL, Cozijnsen AJ, Salisbury PA, Howlett BJ, Van de Wouw AP (2012b) Identifying resistance genes to Leptosphaeria maculans in Australian Brassica napus cultivars based on reactions to isolates with known avirulence genotypes. Crop & Pasture Science 63, 338–350.
Identifying resistance genes to Leptosphaeria maculans in Australian Brassica napus cultivars based on reactions to isolates with known avirulence genotypes.Crossref | GoogleScholarGoogle Scholar |

Mayerhofer R, Bansal VK, Thiagarajah MR, Stringam GR, Good AG (1997) Molecular mapping of resistance to Leptosphaeria maculans in Australian cultivars of Brassica napus. Genome 40, 294–301.
Molecular mapping of resistance to Leptosphaeria maculans in Australian cultivars of Brassica napus.Crossref | GoogleScholarGoogle Scholar |

Mayerhofer R, Wilde K, Mayerhofer M, Lydiate D, Bansal VK, Good AG, Parkin IAP (2005) Complexities of chromosome landing in a highly duplicated genome: Toward map-based cloning of a gene controlling blackleg resistance in Brassica napus. Genetics 171, 1977–1988.
Complexities of chromosome landing in a highly duplicated genome: Toward map-based cloning of a gene controlling blackleg resistance in Brassica napus.Crossref | GoogleScholarGoogle Scholar |

Pang ECK, Halloran GM (1996) The genetics of blackleg [Leptosphaeria maculans (Desm.) Ces. et De Not.] resistance in rapeseed (Brassica napus L.). II. Seedling and adult-plant resistance as quantitative traits. Theoretical and Applied Genetics 93, 941–949.
The genetics of blackleg [Leptosphaeria maculans (Desm.) Ces. et De Not.] resistance in rapeseed (Brassica napus L.). II. Seedling and adult-plant resistance as quantitative traits.Crossref | GoogleScholarGoogle Scholar |

Piquemal J, Cinquin E, Couton F, Rondeau C, Seignoret E, Doucet I, Perret D, Villeger MJ, Vincourt P, Blanchard P (2005) Construction of an oilseed rape (Brassica napus L.) genetic map with SSR markers. Theoretical and Applied Genetics 111, 1514–1523.
Construction of an oilseed rape (Brassica napus L.) genetic map with SSR markers.Crossref | GoogleScholarGoogle Scholar |

R Development Core Team (2011) R: a language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. Avaliable at: www.R-project.org/

Raman R, Raman H, Johnstone K, Lisle C, Smith A, Martin P, Allen H (2005) Genetic and in silico comparative mapping of the polyphenol oxidase gene in bread wheat (Triticum aestivum L.). Functional & Integrative Genomics 5, 185–200.
Genetic and in silico comparative mapping of the polyphenol oxidase gene in bread wheat (Triticum aestivum L.).Crossref | GoogleScholarGoogle Scholar |

Raman R, Allen H, Diffey S, Raman H, Martin P, McKelvie K (2009) Localisation of quantitative trait loci for quality attributes in a doubled haploid population of wheat (Triticum aestivum L.). Genome 52, 701–715.
Localisation of quantitative trait loci for quality attributes in a doubled haploid population of wheat (Triticum aestivum L.).Crossref | GoogleScholarGoogle Scholar |

Raman H, Raman R, et al. (2012a) Genetic and physical mapping of flowering time loci in oilseed rape (Brassica napus L.). Theoretical and Applied Genetics. www.springerlink.com/openurl.asp?genre=article&id=https://doi.org/10.1007/s00122-012-1966-8

Raman H, Raman R, Larkan N (2012b) Molecular dissection of blackleg resistance. In ‘Plant breeding methods’. (Ed. S. B. Andersen) in press. www.intechopen.com/

Raman H, Raman R, et al (2012c) Diversity Array Technology markers: Genetic diversity analyses and linkage map construction in rapeseed (Brassica napus L.). DNA Research 19, 51–65.
Diversity Array Technology markers: Genetic diversity analyses and linkage map construction in rapeseed (Brassica napus L.).Crossref | GoogleScholarGoogle Scholar |

Raman R, Taylor B, et al (2012d) Molecular mapping of qualitative and quantitative loci for resistance to Leptosphaeria maculans; causing blackleg disease in canola (Brassica napus L.). Theoretical and Applied Genetics 125, 405–418.
Molecular mapping of qualitative and quantitative loci for resistance to Leptosphaeria maculans; causing blackleg disease in canola (Brassica napus L.).Crossref | GoogleScholarGoogle Scholar |

Rimmer SR (2006) Resistance genes to Leptosphaeria maculans in Brassica napus. Canadian Journal of Plant Pathology – Revue Canadienne de Phytopathologie 28, S288–S297.
Resistance genes to Leptosphaeria maculans in Brassica napus.Crossref | GoogleScholarGoogle Scholar |

Rimmer SR, Borhan MH, Zhu B, Somers D (1999) Mapping resistance genes in Brassica napus to Leptosphaeria maculans. In ‘Proceedings of the 10th International Rapeseed Congress’. Canberra. (Eds PA Salisbury, TD Potter, G McDonald, AG Green) (Organising Committee of the 10th International Rapeseed Congress/The Regional Institute: Gosford, NSW)

Rouxel T, Penaud A, Pinochet X, Brun H, Gout L, Delourme R, Schmit J, Balesdent M-H (2003a) A 10-year survey of populations of Leptosphaeria maculans in France indicates a rapid adaptation towards the Rlm1 resistance gene of oilseed rape. European Journal of Plant Pathology 109, 871–881.
A 10-year survey of populations of Leptosphaeria maculans in France indicates a rapid adaptation towards the Rlm1 resistance gene of oilseed rape.Crossref | GoogleScholarGoogle Scholar |

Rouxel T, Willner E, Coudard L, Balesdent M-H (2003b) Screening and identification of resistance to Leptosphaeria maculans (stem canker) in Brassica napus accessions. Euphytica 133, 219–231.
Screening and identification of resistance to Leptosphaeria maculans (stem canker) in Brassica napus accessions.Crossref | GoogleScholarGoogle Scholar |

Roy NN (1984) Interspecific transfer of Brassica juncea-type high blackleg resistance to Brassica napus. Euphytica 33, 295–303.
Interspecific transfer of Brassica juncea-type high blackleg resistance to Brassica napus.Crossref | GoogleScholarGoogle Scholar |

Salisbury PA, Wratten N (Eds) (1999) Brassica napus breeding. Canola in Australia: the First 30 Years. In ‘Proceedings of the 10th International Rapeseed Congress’. Canberra. (Eds PA Salisbury, TD Potter, G McDonald, AG Green) pp. 29–35. (Organising Committee of the 10th International Rapeseed Congress/The Regional Institute: Gosford, NSW)

Sjödin C, Glimelius K (1988) Screening for resistance to blackleg Phoma lingam (Tode ex Fr.) Desm. within Brassicaceae. Journal of Phytopathology 123, 322–332.
Screening for resistance to blackleg Phoma lingam (Tode ex Fr.) Desm. within Brassicaceae.Crossref | GoogleScholarGoogle Scholar |

Snowdon RJ, Winter H, Diestel A, Sacristan MD (2000) Development and characterisation of Brassica napus-Sinapus arvensis addition lines exhibiting resistance to Leptosphaeria maculans. Theoretical and Applied Genetics 101, 1008–1014.
Development and characterisation of Brassica napus-Sinapus arvensis addition lines exhibiting resistance to Leptosphaeria maculans.Crossref | GoogleScholarGoogle Scholar |

Sprague SJ, Marcroft SJ, Hayden HL, Howlett BJ (2006) Major gene resistance to blackleg in Brassica napus overcome within three years of commercial production in southeastern Australia. Plant Disease 90, 190–198.
Major gene resistance to blackleg in Brassica napus overcome within three years of commercial production in southeastern Australia.Crossref | GoogleScholarGoogle Scholar |

Suwabe K, Tsukazaki H, Iketani H, Hatakeyama K, Kondo M, Fujimura M, Nunome T, Fukuoka H, Hirai M, Matsumoto S (2006) Simple sequence repeat-based comparative genomics between Brassica rapa and Arabidopsis thaliana: The genetic origin of clubroot resistance. Genetics 173, 309–319.
Simple sequence repeat-based comparative genomics between Brassica rapa and Arabidopsis thaliana: The genetic origin of clubroot resistance.Crossref | GoogleScholarGoogle Scholar |

Tadege M, Sheldon C, Helliwell C, Stoutjesdijk P, Dennis E, Peacock W (2001) Control of flowering time by FLC orthologues in Brassica napus. The Plant Journal 28, 545–553.
Control of flowering time by FLC orthologues in Brassica napus.Crossref | GoogleScholarGoogle Scholar |

Tollenaere R, Hayward A, Dalton-Morgan J, Campbell E, Lee JRM, Lorenc MT, Manoli S, Stiller J, Raman R, Raman H, Edwards D, Batley J (2012) Identification and characterization of candidate Rlm4 blackleg resistance genes in Brassica napus using next-generation sequencing. Plant Biotechnology Journal
Identification and characterization of candidate Rlm4 blackleg resistance genes in Brassica napus using next-generation sequencing.Crossref | GoogleScholarGoogle Scholar |

Trick M, Long Y, Meng J, Bancroft I (2009) Single nucleotide polymorphism (SNP) discovery in the polyploid Brassica napus using Solexa transcriptome sequencing. Plant Biotechnology Journal 7, 334–346.
Single nucleotide polymorphism (SNP) discovery in the polyploid Brassica napus using Solexa transcriptome sequencing.Crossref | GoogleScholarGoogle Scholar |

van Os H, Stam P, Visser R, Eck H (2006) RECORD: a novel method for ordering loci on a genetic linkage map. Theoretical and Applied Genetics 112, 30–40.
RECORD: a novel method for ordering loci on a genetic linkage map.Crossref | GoogleScholarGoogle Scholar |

Voorrips RE (2002) MapChart: Software for the graphical presentation of linkage maps and QTLs. The Journal of Heredity 93, 77–78.
MapChart: Software for the graphical presentation of linkage maps and QTLs.Crossref | GoogleScholarGoogle Scholar |

Wang J, Kaur S, Cogan NOI, Dobrowolski MP, Salisbury PA, Burton WA, Baillie R, Hand M, Hopkins C, Forster JW, Smith KF, Spangerberg G (2009) Assessment of genetic diversity in Australian canola (Brassica napus L.) cultivars using SSR markers. Crop & Pasture Science 60, 1193–1201.
Assessment of genetic diversity in Australian canola (Brassica napus L.) cultivars using SSR markers.Crossref | GoogleScholarGoogle Scholar |

Williams PH (1992) Biology of Leptosphaeria maculans. Canadian Journal of Plant Pathology 14, 30–35.
Biology of Leptosphaeria maculans.Crossref | GoogleScholarGoogle Scholar |

Yu F, Lydiate DJ, Rimmer SR (2005) Identification of two novel genes for blackleg resistance in Brassica napus. Theoretical and Applied Genetics 110, 969–979.
Identification of two novel genes for blackleg resistance in Brassica napus.Crossref | GoogleScholarGoogle Scholar |

Yu F, Lydiate DJ, Rimmer SR (2008) Identification and mapping of a third blackleg resistance locus in Brassica napus derived from B. rapa subsp. sylvestris. Genome 51, 64–72.
Identification and mapping of a third blackleg resistance locus in Brassica napus derived from B. rapa subsp. sylvestris.Crossref | GoogleScholarGoogle Scholar |

Zou X, Suppanz I, Raman H, Hou J, Wang J, Long Y, Jung C, Meng J (2012) Comparative analysis of FLC homologues in Brassicaceae provides insight into their role in the evolution of oilseed rape. PLoS ONE 7, e45751
Comparative analysis of FLC homologues in Brassicaceae provides insight into their role in the evolution of oilseed rape.Crossref | GoogleScholarGoogle Scholar |