Effects of Pyramiding Quantitative Resistance Genes pi21, Pi34, and Pi35 on Rice Leaf Blast Disease.

Development of resistant cultivars has been an effective method for controlling rice blast disease caused by Magnaporthe oryzae. Quantitative blast resistance genes may offer durable resistance because the selection pressure on M. oryzae to overcome resistance is low as a result of the genes' moderate susceptibility. Because the effects of individual resistance genes are relatively small, pyramiding these genes in rice cultivars is a promising strategy. Here, we used near-isogenic and backcross lines of rice cultivar Koshihikari with single- or two-gene combinations of blast resistance genes (pi21, Pi34, and Pi35) to evaluate the suppression of leaf blast. The severity of the disease was assessed throughout the infection process. Resistance varied among the lines: Pi35 conferred the strongest resistance, while Pi34 showed the weakest effects. Two types of combined-gene interactions were observed, and they varied on the basis of gene combination and characteristic of the infection: (i) the combination of two resistance genes was more effective than either of the genes individually or (ii) the combination of two resistance genes was similar to the level of the most effective resistance gene in the pair. The most effective gene combination for the suppression of leaf blast was pi21 + Pi35.

Molecular cloning and characterization of the AVR-Pia locus from a Japanese field isolate of Magnaporthe oryzae.

In order to clone and analyse the avirulence gene AVR-Pia from Japanese field isolates of Magnaporthe oryzae, a mutant of the M. oryzae strain Ina168 was isolated. This mutant, which was named Ina168m95-1, gained virulence towards the rice cultivar Aichi-asahi, which contains the resistance gene Pia. A DNA fragment (named PM01) that was deleted in the mutant and that co-segregated with avirulence towards Aichi-asahi was isolated. Three cosmid clones that included the regions that flanked PM01 were isolated from a genomic DNA library. One of these clones (46F3) complemented the mutant phenotype, which indicated clearly that this clone contained the avirulence gene AVR-Pia. Clone 46F3 contained insertions of transposable elements. The 46F3 insert was divided into fragments I-VI, and these were cloned individually into a hygromycin-resistant vector for the transformation of the mutant Ina168m95-1. An inoculation assay of the transformants revealed that fragment V (3.5 kb) contained AVR-Pia. By deletion analysis of fragment V, AVR-Pia was localized to an 1199-bp DNA fragment, which included a 255-bp open reading frame with weak homology to a bacterial cytochrome-c-like protein. Restriction fragment length polymorphism analysis of this region revealed that this DNA sequence co-segregated with the AVR-Pia locus in a genetic map that was constructed using Chinese isolates.

Novel mating type-dependent transcripts at the mating type locus in Magnaporthe oryzae.

The mating type locus (MAT1) of Magnaporthe oryzae has similar structural organization to MAT in other ascomycetes and encodes the mating type genes MAT1-1-1 with an alpha-box motif and MAT1-2-1 with an HMG-box motif in the MAT1-1 and MAT1-2 idiomorphs, respectively. Sequence and expression analyses of the MAT1 locus indicated a second open reading frame (ORF), MAT1-1-2, in the MAT1-1 idiomorph, and novel mating-type dependent ORFs (MAT1-1-3 and MAT1-2-2) at the locus. The MAT1-1-3 ORF initiated within the MAT1-1 idiomorph while the MAT1-2-2 ORF initiated at the border of the MAT1-2 idiomorph with both ORFs sharing most of their reading frames in the MAT1 flanking region. This suggests that the encoded proteins (MAT1-1-3 and MAT1-2-2) should be similar in their primary structures but can be distinguished by distinct N-termini with amino acids of 1 and 32, respectively, in each mating type. A CT dinucleotide repeat, (CT)n, present in the upstream region of MAT1-1-3, was polymorphic among the isolates.