Abstract
We have used rice line Tetep as a resistant donor with the aim of mapping a durable blast resistance gene Pi-kh using RAPD and AFLP techniques in conjunction with bulk segregant analysis. An F2 mapping population consisting of 205 plants was generated by crossing Tetep with HP2216, a highly susceptible cultivar. Inoculation with specific isolate (PLP-1) of Magnaporthe grisea at seeding stage showed that the Pi-kh gene inherited as a single dominant gene in F2 population. RAPD analysis was performed with 240 primers to detect polymorphism between resistant and susceptible parents. Of these, 48 primers produced polymorphic banding pattern between resistant and susceptible parents. Bulk segregant analysis was performed with 48 primers of which 5 showed polymorphism between resistant and susceptible bulks. A 700 bp DNA band was obtained in resistant F2 plants with primer 5-129 indicating its linkage to the resistance gene. Out of 64 AFLP primer combinations used for polymorphism survey between HP 2216 and Tetep, 11 AFLP primer combinations were able to distinguish the resistant and susceptible bulks. An AFLP band of 75 bp obtained with primer combination, E-TAlM-CTC co-segregated with the resistance gene. The RAPD marker 5-129700 and AFLP75 were placed on the linkage map at a distance of 2.1 eM and 15.1 eM flanking to Pi-khgene, respectively. The RAPD band closely linked to Pi-kh gene was sequenced and used for the development of CAPs markers which also co-segregated with resistant phenotype in the mapping population. On sequence analysis and homology search of RAPD fragment with whole rice genome sequence database and the information available on physical, genetic and sequence maps of rice, the co-segregating CAPs marker was placed at long arm of rice chromosome 11. CAPs marker developed in this study showed polymorphism in different rice cultivars grown in North-Western Himalayan region and is being used for the pyramiding of Pi-kh gene along with other blast resistance genes using marker-assisted selection.
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References
McCouch SR, Nelson RJ, Thome J & Zeigler RS, In Rice blast disease, (RS Zeigler, SA Leong, PS Teng, Editors) CAB International, Wallingford, Oxon, UK (1994) pp 167–187.
Mackill OJ, Bonman JM, Suh HS & Srilingam R, Rice Genet Newsl, 2 (1985) 80. Molecular Mapping of Blast Resistance Gene in Rice 133
Kiyosawa S, Misc Bull Natl Inst Agr Sci, 01 (1974) 1–58.
Sharma TR, Chauhan RS, Singh BM, Paul R, Sagar V & Rathour R, J Phytopath, 150 (2002) 649
Rathour R, Singh BM, Sharma TR & Chauhan RS, J Phytopath, 152 (2004) 304.
Yu ZH, Mackill OJ, Bonman JM & Tanksley SO, Theor ApplGenet, 81 (1991) 471.
Wang G, Mackill OJ, Bonman JM, McCouch SR, Champoux M & Nelson RJ, Genetics, 136 (1994)1421.
Hittalmani S, Foolad MR, Mew T, Rodriguez RL & Haung N, Theor Appl Genet, 91 (1995) 9.
Hittalmani S, Parco A, Mew TW, Zeigler RS & Haung N, Theor Appl Genet, 100 (2000) 1121.
Naqvi NI, Bonman JM, Mackill OJ & Chattoo BB, Mol Breed, 1 (1995) 341.
Miyamoto MI, Rybka, AK, Kodama O & Kawasaki S, Mol Plant-Microbe Interact, 9 (1996) 6.
Vos P, Hogers R, Bleeker M, Rijans M, Lee T van der, Homes M, Frijters A, Plot J, Peleman J, Kuiper M & Zabeau M, Nucl Acids Res, 23 (1995) 4407.
Chen OH, Oela Vina M, Inukai T, Mackill OJ, Ronald PC & Nelson RJ, Theor Appl Genet, 98 (1999) 1046.
Farman ML & Leong SA, Genetics, 150 (1998) 1049.
Bonman JM, Vergel OOTI & Khin MM, Plant Dis, 70 (1986) 767.
Murray MG & Thompson WF, Nucl Acids Res, 8 (1980) 4321.
Williams JGK, Kubelik A, Livak K, Rafalski J & Tingey SV, Nucl Acids Res, 18 (1990) 6531.
Lander ESP, Green J, Abhrahamson A & Barlow MJ, Genomics, 1 (1987) 174.
Altschul SF, Gisi W, Miller W, Myers EW & Lipman OJ, J Mol Biol, 215 (1990) 403.
Mackill OJ & Bonman JM, Phytopathology, 82 (1992) 746.
Inukai T, Nelson RJ, Zeigler RS, Sarkarung S, Mackill OJ, Bonman JM, Takamura I & Kinoshita T, Phytopathology, 84 (1994) 1278.
Michelmore RW, Paran I & Kessli V, Proc Natl Acad Sci, USA, 88 (1991) 9824
Bryan GT, Wu KS, Farrall L, Jia Y, Hershey HP, McAdams SA, Faulk KN, Oonaldsen GK, Trachini R & Valent B, Plant Cell, 12 (2000) 2033.
Yu ZH, Mackill OJ, Bonman JM, McCouch S, Guiderdoni E, Noteghem, JL & Tanksley SO, Theor Appl Genet, 93 (1996) 859.
Paran I & Michelmore RN, Theor Appl Genet, 85 (1993) 985.
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Sharma, T.R., Shanker, P., Singh, B.K. et al. Molecular Mapping of Rice Blast Resistance Gene Pi-kh in the Rice Variety Tetep. J. Plant Biochem. Biotechnol. 14, 127–133 (2005). https://doi.org/10.1007/BF03263240
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DOI: https://doi.org/10.1007/BF03263240