Breeding of Rice Restorer Lines Carrying Blast Resistance Gene Pi25 by MarkerAssisted Selection
ZHOU Haipeng1,#, ZHAN Xiaodeng1,#, CAI Rongyao2, CHENG Shihua1,*, CAO Liyong1,*
1Chinese National Center for Rice Improvement/State Key Laboratory of Rice Biology, China National Rice Research Institute, Hangzhou 310006, China；2Institute of Plant Protection and Microbiology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China； #These authors contributed equally to this paper; *Corresponding authors, E-mail: firstname.lastname@example.org；email@example.com
Three recombinant inbred lines (RILs) carrying rice blast resistance gene Pi25 from the combination Zhong 156/Gumei 2 were crossed with highyielding restorer lines, 9308 and 311. The seeds were collected and advanced by single seed descent, and six RIL populations at the F6 generation were obtained and used for the study. Based on the previously reported genetic map, four additional SSR markers RM3330, RM564, RM3628 and RM6298 were added to the interval harboring Pi25 on rice chromosome 6. Relied on the marker genotyping of RM3330 and A7, 109 Pi25 homozygous lines were selected from the six RIL populations. Markerassisted selection analysis was carried out on the acquired 109 Pi25 homozygous lines using existing markers linked to fertility restoring genes. As a result, twenty restorer lines resistant to blast were obtained. Studies on fertility restoring ability of the 20 restorer lines concluded that the seedsetting rate of crosses between these restorer lines and cytoplasmic male sterility lines (Zhong 9A and Ⅱ32A) reached a normal level. In addition, three populations were selected to be artificially inoculated with a regionally prevalent isolate 05201 of Magnaporthe grisea to evaluate the accuracy of markerassisted selection. The result manifested that simultaneous selection of the markers flanking the gene on both sides could increase the selection accuracy significantly.
ZHOU Haipeng,#,ZHAN Xiaodeng et al. Breeding of Rice Restorer Lines Carrying Blast Resistance Gene Pi25 by MarkerAssisted Selection[J]. , 2008, 22(6): 590-590～596 .
Ou S H. Rice Disease. Farnham Royal, UK: Commonwealth Agricultural Bureaux, 1985.Yu Z H, Mackill D J, Bonman J M, et al. Molecular mapping of genes for resistance to rice blast (Pyricularia grisea Sacc.). Theor Appl Genet, 1996, 93: 859-863.Yu Z H, Mackill D J, Bonman J M, et al. Tagging genes for blast resistance in rice via linkage to RFLP markers. Theor Appl Genet, 1991, 81: 471-476. Wang G L, Mackill D J, Bonman M, et al. RFLP mapping of genes conferring complete and partial resistance to blast in a durably resistant rice cultivar. Genetics, 1994, 136: 1421-1434.Naqvi N I, Chattoo B B. Development of a sequence characterized amplified region (SCAR) based indirect selection method for a dominant blastresistance gene in rice. Genome, 1996, 39: 26-30.Zhuang J Y, Ma W B， Wu J L, et al. Mapping of leaf and neck blast resistance genes with resistance gene analog, RAPD and RFLP rice. Euphytica, 2002, 128(3): 363-370.吴建利, 庄杰云, 柴荣耀, 等．水稻抗穗瘟基因的分子定位．植物病理学报, 2000, 30(2): 111-115．吴建利, 柴荣耀, 樊叶杨, 等．抗稻瘟病水稻材料谷梅2号中主效抗稻瘟病基因的成簇分布．中国水稻科学, 2004, 18(6): 567-569．Wu J L, Fan Y Y, Li D D, et al. Genetic control of rice blast resistance in the durably resistant cultivar Gumei 2 against multiple isolates. Theor Appl Genet, 2005, 111: 50-56.卢扬江, 郑康乐．提取水稻DNA 的一种简易方法．中国水稻科学, 1992, 6(1): 47-48．柴荣耀, 金敏忠. 水稻稻瘟病离体接种技术研究初报．浙江农业学报, 1995, 7(6): 488.Lander E S, Green P, Abrahamson J, et al. MAPMAKER: An interactive computer program for constructing primary genetic maps of experimental and natural populations. Genomics, 1987, 1: 174-181. Kosambi D D. The estimation of map distances from recombination values. Ann Eugen, 1944, 12: 172-175.Causse M A, Fulton T M, Cho Y G, et al. Saturated molecu1ar map of the rice genome based on an interspecific backcross population. Genetics, 1994, 138: 1251-1274.Pan Q H, Wang L, Ikehashi H, et al. Identification of a new blast resistance gene in the indica cultivar Kasalath using Japanese differential cultivars and isozyme markers. Phytopathology, 1996, 86: 1071-1075.Liu G, Lu G, Zeng L, et al. Two broadspectrum blast resistance genes, Pi9(t) and Pi2(t) are physically linked on rice chromosome 6. Mol Genet Gen, 2002, 267: 472-480.Yi N. A unified Markov chain Monte Carlo framework for mapping multiple quantitative trait loci. Genetics, 2004, 167: 967-975.Ni J, Colowit P M, Oster J J, et al. Molecular markers linked to stem rot resistance in rice. Theor Appl Genet, 2001, 102: 511-516.ConawayBormans C A, Marchetti M A, Johnson C W, et al. Molecular markers linked to the blast resistance gene Piz in Oryza sativa for use in marker assisted selection. Theor Appl Genet, 2003, 107(6): 1014-1020.Narayanan N N, Baisakh N, Oliva N P, et al. Molecular breeding: Markerassisted selection combined with biolistic transformation for blast and bacterial blight resistance in indica rice (cv. CO39). Mol Breeding, 2004, 14: 61-71.金素娟， 柳武革， 朱小源， 等. 利用分子辅助选择改良温敏核不育系GD8S的稻瘟病抗性. 中国水稻科学， 2007, 21（6）： 599-604.Huang N, Angels E R, Domingo J, et al. Pyramiding of bacterial blight resistance genes in rice: Markerassisted selection using RFLP and PCR. Theor Appl Genet, 1997, 95: 313-320.Cao L Y, Zhuang J Y, Yuan S J, et al. Hybrid rice resistance to bacterial leaf blight developed by marker assisted selection. Rice Sci, 2003, 11: 68-70.