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2004; Xiang et?al. 2006). The 60 F2 plants did not carry Xa3/Xa26 based on PCR amplification using gene-specific primers (Supporting Information Table?S1) and were heterozygous at markers RM179 and RM28172, which harboured xa25/Xa25(t) at chromosome 12 (Chen et?al. 2002). For confirmation of fine mapping results, 20 progenies (F4) of each of 12 recombinant F3 plants were further examined for their http://www.selleckchem.com/products/Vorinostat-saha.html responses to Xoo infection. Another F2 plant, S6 that did not carry Xa3/Xa26 based on PCR amplification using gene-specific primers (Supporting Information Table?S1) and homozygous as Minghui 63 at markers RM179 and RM28172, was identified. S6 was resistance to Xoo strain PXO339 as Minghui 63. The next generation of S6, S6-1, was used to represent Minghui 63 at xa25/Xa25(t) locus to cross with susceptible Zhenshan 97 for further analysis. A sample of 255 random F3 plants from the mapping population was used for mapping the target gene. A molecular linkage map was constructed using the computer program MAPMAKER/EXP 3.0 (Lincoln, Daly & Lander 1992). The maximum-likelihood map order for markers and the chromosomal location of the target gene on the molecular linkage map were determined with a logarithm of odds threshold of 3.0. The genetic distance [centimorgans (cM)] between markers for fine mapping of the target gene using 795 highly resistant F3 plants (average lesion length? http://www.selleck.cn/products/ve-821.html number of the highly resistant plants surveyed, N1 is the number of plants homozygous for the marker genotype of resistance Minghui 63, and N2 is the number of plants heterozygous http://www.selleckchem.com/products/Cisplatin.html for the two parents. To evaluate bacterial blight disease, plants were inoculated with Philippine Xoo strains, PXO61 (race 1), PXO99 (race 6), PXO339 (race 9) or PXO341 (race 10) at the seedling (four- to six-leaf) or adult [booting (panicle development)] stage by the leaf-clipping method (Chen et?al. 2002). Because the parents of the mapping population, Minghui 63 and Zhenshan 97, had different lengths of life cycle and the F3 individuals in the population could not grow to booting stage at the same time, plants in the population were inoculated with Xoo at four- to six-leaf stage. Disease was scored by measuring the lesion length (cm) or the percent lesion area (lesion length/leaf length) at 2?weeks after inoculation. For analysing gene expression in response to Xoo infection, the 2 to 3?cm leaf fragments next to bacterial infection sites were used for RNA isolation. The growth rate of Xoo in rice leaves was determined from three leaves at each time point by counting colony-forming units (cfu; Sun et?al. 2004). Leaves were sterilized using 75% ethanol, ground separately, suspended in sterilized water and plated on potato agar media. To transfer the dominant Xa25 gene, a 5.2?kb DNA fragment including 1.