Abstract
The interaction between rice and the rice blast fungus Magnaporthe oryzae follows a gene-for-gene model. The pathotype of a strain is determined by its avirulence gene content. In this study, we crossed avirulent strain S1522 and virulent strain S159 to generate 108 progenies. We subsequently isolated the avirulence gene Avr-Pik m through classical genetic analysis. The segregation ratio in F1 population was 1:1 and confirmed the presence of a single locus in the genome of S1522. Two SCAR and five SSR markers linked to the avirulence gene were identified from the segregated population, and Avr-Pik m was mapped on the chromosome of M. oryzae. A genomic library of avirulence parental strain S1522 was then constructed; the library was screened using the markers SCE121406 and SSR47A18, which were linked to the AVR gene as probes. Using these procedures, a fine physical map was assembled to include five TAC clones. TAC clone 35C5 is 32 kb in length and contains the two above-mentioned SCE121406 and SSR47A18 probes, suggesting that the Avr-Pik m gene spans across the two markers located on the clone. These results provide support towards Avr-Pik m map-based cloning.
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Bishop AJR, Louis EJ, Borts RH (2000) Minisatellite variants generated in yeast meiosis involve DNA removal during gene conversion. Genetics 156:7–20
Bonert HU, Fudal I, Dioh W, Tharreau D, Notteghem JL, Lebrun MH (2004) A putative polyketide synthase/peptide synthetase from Magnaporthe grisea signals pathogen attack to resistant rice. Plant Cell 16:2499–2513
Bonman JM (1992) Durable resistance to rice blast disease-environmental influences. Euphytica 63:115–123
Chauhan RS, Farman ML, Zhang HB, Leong SA (2002) Genetic and physical mapping of a rice blast resistance locus, Pi-CO39(t), that corresponds to the avirulence gene AVR1-CO39 of Magnaporthe grisea. Mol Genet Genom 267:603–612
Chen QH, Wang YC, Zheng XB (2006) Genetic analysis and molecular mapping of the avirulence gene PRE1, a gene for host-species specificity in the blast fungus Magnaporthe grisea. Genome 49:873–881
Chen QH, Wang YC, Li AN, Zhang ZG, Zheng XB (2007) Molecular mapping of two cultivar-specific avirulence genes in the rice blast fungus Magnaporthe grisea. Mol Genet Genom 277:139–148
Dean RA, Talbot NJ, Ebbole DJ, Farman ML, Mitchell TK, Orbach MJ, Thon M, Kulkarni R, Xu JR, Pan H, Read ND, Lee YH, Carbone I, Brown D, Oh YY, Donofrio N, Jeong JS, Soanes DM, Djonovic S, Kolomiets E, Rehmeyer C, Li W, Harding M, Kim S, Lebrun MH, Bohnert H, Coughlan S, Butler J, Calvo S, Ma LJ, Nicol R, Purcell S, Nusbaum C, Galagan JE, Birren BW (2005) The genome sequence of the rice blast fungus Magnaporthe grisea. Nature 434:980–986
Dioh W, Tharreau D, Notteghem JL, Orbach M, Lebrun MH (2000) Mapping of avirulence genes in the rice blast fungus, Magnaporthe grisea, with RFLP and RAPD markers. Mol Plant Microbe Interact 13:217–227
Farman ML, Leong SA (1998) Chromosome walking to the AVR1-CO39 avirulence gene of Magnaporthe grisea: discrepancy between the physical and genetic maps. Genetics 150:1049–1058
Flor HH (1971) Current status of the gene-for-gene concept. Annu Rev Phytopathol 9:275–296
Gao W, Khang CH, Park SY, Lee YH, Kang S (2002) Evolution and organization of a highly dynamic, subtelomeric helicase gene family in the rice blast fungus Magnaporthe grisea. Genetics 162:103–112
Graham GC, Mayers P, Henry RJ (1994) A simplified method for the preparation of fungal genomic DNA for PCR and RAPD analysis. Biotechniques 16:48–50
Hamer JE, Farrall L, Orbach MJ, Valent B, Chumley FG (1989) Host species-specific conservation of a family of repeated DNA sequences in the genome of a fungal plant pathogen. Proc Natl Acad Sci USA 86:9981–9985
Hirayae K, Miyasaka A, Miyazaki C, Hayashi N, Naito H, Nishi K, Iwano M (1999) Screening of AFLP markers for the construction of molecular linkage map in Magnaporthe grisea. Ann Phytopathol Soc Jpn 65:342 (In Japanese with English title)
Kang S, Sweigard JA, Valent B (1995) The PWL host specificity gene family in the blast fungus Magnaporthe grisea. Mol Plant Microbe Interact 8:939–948
Kaye C, Milazzo J, Rozenfeld S, Lebrun MH, Tharreau D (2003) The development of simple sequence repeat markers for Magnaporthe grisea and their integration into an established genetic linkage map. Fungal Genet Biol 40:207–214
Kiyosawa S (1982) Genetics and epidemiological modeling of breakdown of plant disease resistance. Annu Rev Phytopathol 20:93–117
Leach JE, White FF (1996) Bacterial avirulence genes. Annu Rev Phytopathol 34:153–179
Li CY, Luo CX, Li JB, Shen Y, Ise K (2000) Mapping avirulence gene in the rice blast fungus Magnaporthe grisea. Sci Agric Sin 33:49–53 (in Chinese with English summary)
Lin F, Li JB, Li CY, Wang L, Pan QH (2002) Identification of two avirulence genes in the rice blast fungus Magnaporthe grisea using Random Amplified Polymorphic DNA (RAPD) markers (in Chinese with English summary). Sci Agric Sin 35:1079–1083
Luo CX, Fujita Y, Yasuda N, Hirayae K, Nakajima T, Hayashi N, Kusaba M, Yaegashi H (2004) Identification of Magnaporthe oryzae avirulence genes to three rice blast resistance genes. Plant Dis 88:265–270
Ma JH, Wang L, Feng SJ, Lin F, Xiao Y, Pan QH (2006) Identification and fine mapping of AvrPi15, a novel avirulence gene of Magnaporthe grisea. Theor Appl Genet 113:875–883
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 88:9828–9832
Nitta N, Farman ML, Leong SA (1997)Genome organization of Magnaporthe grisea: integration of genetic maps, clustering of transposable elements and identification of genome duplications and rearrangements. Theor Appl Genet 95:20–32
Notteghem JL, Tharreau D, Silue D, Roumen E (1994) Present knowledge of rice resistance genetics and strategies for Magnaporthe grisea pathogenicity and avirulence gene analysis. Rice blast disease. CAB International, Wallingford, pp 155–165
Orbach MJ, Farrall L, Sweigard JA, Chumley FG, Valent B (2000) A telomeric avirulence gene determines efficacy for the rice blast resistance gene Pi-ta. Plant Cell 12:2019–2032
Ou SH (1980) Pathogen variability and host resistance in rice blast disease. Annu Rev Phytopathol 18:167–187
Ou SH (1985) Rice diseases, 2nd edn. Commonwealth Mycological Institute, Kew, Surrey, pp 109–201
Peng YL, Shishiyama J (1988) Temporal sequence of cytological events in rice leaves infected with Pyricularia oryzae. Can J Bot 66:730–735
Rossman AY, Howard RJ, Valent B (1990) Pyricularia grisea, the correct name for the rice blast disease fungus. Mycologia 82:509–512
Sweigard JA, Valent B, Orbach MJ, Walter AM, Rafalski A, Chumley FG (1993) A genetic map of the rice blast fungus Magnaporthe grisea. In: O’Brien SJ (ed) Genetic maps, 6th edn. Cold Spring Harbor Laboratory, Cold Spring Harbor, pp 3112–3117
Sweigard JA, Carroll AM, Kang S, Farrall L, Chumley FG, Valent B (1995) Identification, cloning, and characterization of PWL2, a gene for host species specificity in the rice blast fungus. Plant Cell 7:1221–1233
Valent B (1990) Rice blast as a model system for plant pathology. Phytopathology 80:33–36
Valent B, Chumley FG (1991) Molecular genetic analysis of the rice blast fungus, magnaporthe grisea. Annu Rev Phytopathol 29:443–467
Wang BH, Lu GD, Lin WM, Wang ZH (2002) Genetic analysis and molecular marker of Avr-Pi1, Avr-Pi2 and Avr-Pi4a of Magnaporthe grisea. Acta Genet Sin 29:820–826
Wei J, Goldberg MB, Burland V, Venkatesan MM, Deng W, Fournier G, Mayhew GF, Plunkett G 3rd, Rose DJ, Darling A, Mau B, Perna NT, Payne SM, Runyen-Janecky LJ, Zhou S, Schwartz DC, Blattner FR (2003) Complete genome sequence and comparative genomics of Shigella flexneri serotype 2a strain 2457T. Infect Immun 71:2775–2786
Yasuda N, Fujita Y, Noguchi M (2004) Identification of avirulence genes in the rice blast fungus corresponding to three resistance genes in Japanese differentials. J Gen Plant Pathol 70:202–206
Yoshida K, Saitoh H, Fujisawa S, Kanzaki H, Matsumura H, Yoshida K, Tosa Y, Chuma I, Takano Y, Win J (2009) Association genetics reveals three novel avirulence genes from the rice blast fungal pathogen Magnaporthe oryzae. Plant Cell 21:1573–1591
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The work was supported by 973 Grant 2006CB101901 and 2012CB114002 (to You-Liang Peng)
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Jiye Yan and Lianhong Zhang contributed equally to the paper
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Yan, J., Zhang, L., Zhao, W. et al. Genetic and physical mapping of the avirulence gene Avr-Pik m in Magnaporthe oryzae . Ann Microbiol 63, 997–1004 (2013). https://doi.org/10.1007/s13213-012-0553-1
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DOI: https://doi.org/10.1007/s13213-012-0553-1