Ryegrass blast, also called gray leaf spot, is caused by the fungus Pyricularia sp. It is one of the most serious diseases of Italian ryegrass (Lolium multiflorum Lam.) in Japan. We analyzed segregation of resistance in an F1 population from a cross between a resistant and a susceptible cultivar. The disease severity distribution in the F1 population suggested that resistance was controlled by a major gene (LmPi1). Analysis of amplified fragment length polymorphisms with bulked segregant analysis identified several markers tightly linked to LmPi1. To identify other markers linked to LmPi1, we used expressed sequence tag-cleaved amplified polymorphic sequence (EST-CAPS) markers mapped in a reference population of Italian ryegrass. Of the 30 EST-CAPS markers screened, one marker, p56, flanking the LmPi1 locus was found. The restriction pattern of p56 amplification showed a unique fragment corresponding to the resistant allele at the LmPi1 locus. A linkage map constructed from the reference population showed that the LmPi1 locus was located in linkage group 5 of Italian ryegrass. Genotype results obtained from resistant and susceptible cultivars indicate that the p56 marker is useful for introduction of the LmPi1 gene into susceptible germplasm in order to develop ryegrass cultivars with enhanced resistance to ryegrass blast.
This is a preview of subscription content, log in to check access.
Buy single article
Instant access to the full article PDF.
Price includes VAT for USA
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
This is the net price. Taxes to be calculated in checkout.
Bert PF, Charmet G, Sourdille P, Hayward MD, Balfourier F (1999) A high-density molecular map for ryegrass (Lolium perenne) using AFLP markers. Theor Appl Genet 99:445–452
Curley J, Sim SC, Jung G, Leong S, Warnke S, Barker RE (2004) QTL mapping of ryegrass blast resistance in ryegrass, and synteny-based comparison with rice blast resistance genes in rice. In: Hopkins A, Wang ZY, Mian R, Sledge M, Barkers RE (eds) Molecular breeding of forage and turf. Kluwer, Dordrecht, pp 37–46
Dumsday JL, Smith KF, Forster JW, Jones ES (2003) SSR-based genetic linkage analysis of resistance to crown rust (Puccinia coronata f. sp. lolii) in perennial ryegrass (Lolium perenne) Plant Pathol 52:628–637
Forster JW, Jones ES, Kölliker R, Drayton MC, Dumsday J, Dupal MP, Guthridge KM, Mahoney NL, van Zijll de Jong E, Smith KF (2001) Development and implementation of molecular markers for forage crop improvement. In: Spangenberg G (ed) Molecular breeding of forage crops. Kluwer, Dordrecht, pp 101–133
Fujimori M, Hirata M, Sugita S, Inoue M, Cai H, Akiyama F, Mano Y, Komatsu T (2000) Development of a high density map in Italian ryegrass (Lolium multiflorum Lam.) using amplified fragment length polymorphism. In: 2nd international symposium on molecular breeding of forage crops, 19–24 November 2000, Lorne and Hamilton, Victoria, Australia
Fujimori M, Hayashi K, Hirata M, Mizuno K, Fujiwara T, Akiyama F, Mano Y, Komatsu T, Takamizo T (2003) Linkage analysis of crown rust resistance gene in Italian ryegrass (Lolium multiflorum Lam.). In: Proceedings of plant and animal genomes XI conference, 11–15 January 2003, San Diego, p 46
Grattapaglia D, Sederoff R (1994) Genetic linkage maps of Eucalyptus grandis and Eucalyptus urophylla using a pseudo-testcross: mapping strategy and RAPD markers. Genetics 137:1121–1137
Hayward MD, Mcadam NJ, Jones JG, Evans C, Evans GM, Forster JM, Ustin A, Hossain KG, Quader B, Stammers M, Will JK(1994) Genetic markers and the selection of quantitative traits in forage grasses. Euphytica 77:269–275
Hayward MD, Forster JW, Jones JG, Dolstra O, Evans C, McAdam NJ, Hossain KG, Stammers M, Will J, Humphreys MO, Evans GM (1998) Genetic analysis of Lolium. I. Identification of linkage groups and the establishment of a genetic map. Plant Breed 117:451–455
Hirata M, Fujimori M, Komatsu T (2000) Development of simple sequence repeat (SSR) markers in Italian ryegrass. In: 2nd international symposium on molecular breeding of forage crops, 19–24 November 2000, Lorne and Hamilton, Victoria, Australia
Ikeda S, Takahashi W, Oishi H (2004) Generation of expressed sequence tags from cDNA libraries of Italian ryegrass (Lolium multiflorum Lam.). Grassland Sci 49:593–598
Inoue M, Gao Z, Hirata M, Fujimori M, Cai H (2004) Construction of a high-density linkage map of Italian ryegrass (Lolium multiflorum Lam.) using restriction fragment length polymorphism, amplified fragment length polymorphism, and telomeric repeat associated sequence markers. Genome 47:57–65
Jones ES, Dupal MD, Dumsday JL, Hughes LJ, Forster JW (2002a) An SSR-based genetic linkage map for perennial ryegrass (Lolium perenne). Theor Appl Genet 102:405–415
Jones ES, Mahoney NL, Hayward MD, Armstead IP, Jones JG, Humphreys MO, King IP, Kishida T, Yamada T, Balfourier F, Charmet G, Forster JW (2002b) An enhanced molecular marker based genetic map of perennial ryegrass (Lolium perenne) reveals comparative relationships with other Poaceae genomes. Genome 45:282–295
Kosambi DD (1944) The estimation of map distances from recombination values. Ann Eugen 12:172–175
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 genome regions by using segregating populations. Proc Natl Acad Sci USA 88:9828–9832
Mizuno K, Yokohata Y, Oda T, Fujiwara T, Hayashi K, Ozaki R, Kobashi K, Ashizawa H, Ushimi T (2003) Breeding of a new variety ‘Sachiaoba’ in Italian ryegrass (Lolium multiflorum L.) and its characteristics (in Japanese with English summary). Bull Yamaguchi Agric Expt Stn 54:11–24
Møller MG, Taylor C, Rasmussen SK, Holm PB (2003) Molecular cloning and characterization of two genes encoding asparagine synthetase in barley (Hordeum vulgare L.). Biochim Biophys Acta 1628:123–132
Murray M, Thompson WF (1980) Rapid isolation of high-molecular-weight plant DNA. Nucleic Acids Res 8: 4321–4325
Myburg AA, Remington DL, O’Malley DM, Sederoff RR, Whetten RW (2001) High-throughput AFLP analysis using infrared dye-labeled primers and an automated DNA sequencer. BioTechniques 30:348–357
Sumida Y, Fukubara H, Inoue T, Kajihara H, Mizuno K, Fujiwara T (2003) A study of the occurrence of dieback on Italian ryegrass (Lolium multiflorum) caused by Pyricularia sp. —Investigation of facts about occurrence and development of a new resistance test method on breeding (in Japanese with English summary). Bull Yamaguchi Agric Exp Stn 54:29–36
Tabien RE, Li Z, Paterson AH, Marchetti MA, Stansel JW, Pinson SRM (2002) Mapping QTLs for field resistance to the rice blast pathogen and evaluating their individual and combined utility in improved varieties. Theor Appl Genet 105:313–324
Van Ooijen JW, Maliepaard C (1996) MapQTL version 3.0 software for the calculation of QTL positions on genetic maps. Plant Research International, Wageningen, the Netherlands
Van Ooijen JW, Voorrips RE (2001) JoinMap 3.0 software for the calculation of genetic linkage maps. Plant Research International, Wageningen, the Netherlands.
Vos P, Hogers R, Bleekers M, Reijans M, van der Lee T, Hornes M, Frijters A, Pot J, Peleman J, Kuiper M, Zabesu M (1995) AFLP: a new technique for DNA fingerprinting. Nucleic Acids Res 23:4407–4414
Xu ML, Melchinger AE, Xia XC, Lübberstedt T (1999) High-resolution mapping of loci conferring resistance to sugarcane mosaic virus in maize using RFLP, SSR, and AFLP markers. Mol Gen Genet 261:574–581
We would like to thank Y. Sumida, H. Kajihara, and K. Nishimi (Yamaguchi Prefecture Agricultural Experiment Station) for providing ryegrass blast fungus and for suggestions on the inoculation test. We are grateful to S. Sato, K. Suginobu, and S. Yazaki (GAFSA) for their good advice and to E. Kobayashi (GAFSA) for her technical assistance. This study was financially supported by a grant from the Japan Racing Association.
Communicated by T. Sasaki
About this article
Cite this article
Miura, Y., Ding, C., Ozaki, R. et al. Development of EST-derived CAPS and AFLP markers linked to a gene for resistance to ryegrass blast (Pyricularia sp.) in Italian ryegrass (Lolium multiflorum Lam.). Theor Appl Genet 111, 811–818 (2005). https://doi.org/10.1007/s00122-005-0001-8
- Amplify Fragment Length Polymorphism
- Perennial Ryegrass
- Amplify Fragment Length Polymorphism Marker
- Cleave Amplify Polymorphic Sequence
- Resistant Parent