Characterization and fine mapping of the rice blast resistance gene Pia

Blast,caused by Magnaporthe oryzae,is one of the most widespread and destructive diseases of rice.Breeding durable resistant cultivars(cvs) can be achieved by pyramiding of various resistance(R) genes.Pia,carried by cv.Aichi Asahi,was evaluated against 612 isolates of M.oryzae collected from 10 Chinese provinces.The Pia gene expresses weak resistance in all the provinces except for Jiangsu.Genomic position-ready marker-based linkage analysis was carried out in a mapping population consisting of 800 F2 plants derived from a cross of Aichi Asahi×Kasalath.The locus was defined in an interval of approximately 90 kb,flanked by markers A16 and A21.Four candidate genes(Pia-1,Pia-2,Pia-3,and Pia-4) ,all having the R gene conserved structure,were predicted in the interval using the cv.Nipponbare genomic sequence.Four candidate resistance gene(CRG) markers(A17,A25,A26,and A27) ,derived from the four candidates,were subjected to genotyping with the recombinants detected at the flanking markers.The first three markers completely co-segregated with the Pia locus,and the fourth was absent in the Aichi Asahi genome and disordered with the Pia locus and its flanking markers,indicating that the fourth candidate gene,Pia-4,could be excluded.Co-segregation marker-based genotyping of the three sets of differentials with known R gene genotypes revealed that the genotype of A26(Pia-3) perfectly matched the R gene genotype of Pia,indicating that Pia-3 is the strongest candidate gene for Pia.