Polymorphism analysis of genomic regions associated with broad-spectrum effective blast resistance genes for marker development in rice
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Cultivated European rice germplasm is generally characterized by moderate to high sensitivity to blast, and blast resistance is therefore one of the most important traits to improve in rice breeding. We collected a panel of 25 rice genotypes containing 13 broad range rice resistance genes that are commonly used in breeding programs around the world: Pi1, Pi2, Pi5, Pi7, Pi9, Pi33, Pib, Pik, Pik-p, Pita, Pita 2 , Piz and Piz-t. The efficiency of the selected Pi genes towards Italian blast pathotypes was tested via artificial inoculation and under natural field infection conditions. To characterize haplotypes present in the chromosomal regions of the blast resistance genes, a polymorphism search was conducted in the sequence regions adjacent to the blast resistance by examining DNA from the Pi gene donors with a panel of 5–7 potential receivers (cultivated European rice genotypes). Seven InDel and 8 presence/absence polymorphisms were directly detected by gel analysis after DNA amplification, while sequencing of 12.870 bp through 32 loci in different genotypes revealed 85 SNP (one SNP every 151 bp). Seven SSRs were additionally tested revealing 5 polymorphic markers between donors and receivers. Polymorphisms were used to develop 35 PCR-based molecular markers suitable for introgressing of Pi genes into a set of the European rice germplasm. For this last purpose, allelic molecular marker variation was evaluated within a representative collection of about 95 rice genotypes. Polymorphic combinations allowing introgression of the broad spectrum resistance genes into a susceptible genetic background have been identified, thus confirming the potential of the identified markers for molecular-assisted breeding.
KeywordsRice (Oryza sativa) Blast (Magnaporthe oryzae) resistance Haplotype analysis Marker-assisted selection (MAS) Gene pyramiding
The authors wish to thank the following persons for the gift of rice genotypes and blast isolates: Dr. Harold Bockelman (USDA, Agricultural Research Service, National Small Grains Research Facility, National Small Grains Collection, USA) for C101LAC, Bala, Carreon, Te Qing, Saber, Katy, Kaybonnet, Zenith, Jefferson; Dr. Didier Tharreau (UMR BGPI, CIRAD, France) for C101LAC, IR64, IR1529, K1, K60 and for the three Italian blast strains; Dr. Jong-Seong Jeon (Graduate School of Biotechnology & Plant Metabolism Research Center Kyung Hee University Korea) for RIL260, IRBL5-M, IRBL7-M; Dr. Kazutoshi Okuno (National Institute of Agrobiological Sciences NIAS, Japan) for Kanto 51, Kusabue, BL1, K1, Fukunishiki, Toride 1 and Dr. Guo-Liang Wang (Department of Plant Pathology, The Ohio State University, USA) for 75-1-127 and C101A-51. This study acknowledges financial support of the project VALORYZA (Italian Ministero per le Politiche Agricole e Agroalimentari, DM 301/7303/06), CEREALAB and SITEIA (Italian Region Emilia Romagna) and of the EU co-funded project EURIGEN (049 AGRI GEN RES).
- 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 434:980–986CrossRefPubMedGoogle Scholar
- Fukuta Y, Telebanco-Yanoria MJ, Imbe T, Tsunematsu H, Kato H, Ebron LA, Hayashi N, Ando I, Khush GS (2003) Development of new international standard differential variety series to integrate blast resistance of rice (Oryza sativa L). JIRCAS Res Highlights 2003:16–17Google Scholar
- Kiyosawa S (1972) Genetics of blast resistance. In: Rice breeding IRRI. Manila, Phillipines pp 203–225Google Scholar
- Lee SK, Song MY, Seo YS, Kim HK, Ko S, Cao PJ, Suh JP, Yi G, Roh JH, Lee S, An G, Hahn TR, Wang GL, Ronald P, Jeon JS (2009) Rice Pi5-mediated resistance to Magnaporthe oryzae requires the presence of two coiled-coil-nucleotide-binding-leucine-rich repeat genes. Genetics 181:1627–1638CrossRefPubMedGoogle Scholar
- Picco AM, Rodino D, Rodolfi M, Sala F (2001) Biologia di Pyricularia grisea (Cooke) Saccardo. http://www agricoltura regione lombardia it/admin/rla_Documenti/1-1235/biologia_di_p grisea_ok pdf
- Scardaci SC, Webster RK, Greer JE, Hill JF, Williams JF, Mutters RG, Brandon DM, McKenzie KS, Oster JJ (1997) Rice blast: a new disease in California. http://www plantsciences ucdavis edu/uccerice/AFS/agfs0297 htm
- Sere Y, Onasanya A, Afolabi A, Mignouna HD, Akator K (2007) Genetic diversity of the blast fungus, Magnaporthe grisea (Hebert) Barr, in Burkina Faso. African J Biotechnol 6:2568–2577Google Scholar
- Sivaraj R, Gnanamanickam SS, Levy M (1996) Pyricularia grisea: a molecular approach for management of rice blast, vol 132. Rice Genetics III International Rice Research Institute, Manila, pp 958–962 Khush GS (ed)Google Scholar
- Staden R, Beal KF, Bonfield JK (1999) The staden package, 1998. Meth Mol Biol 132:115–130Google Scholar
- Zhou B, Qu S, Liu G, Dolan M, Sakai H, Lu G, Bellizzi M, Wang GL (2006) The eight amino-acid differences within three leucine-rich repeats between Pi2 and Piz-t resistance proteins determine the resistance specificity to Magnaporthe grisea. Mol Plant Microbe Interact 19:1216–1228CrossRefPubMedGoogle Scholar