Identification and Fine Mapping of Pi69(t), a New Gene Conferring Broad-Spectrum Resistance Against Magnaporthe oryzae From Oryza glaberrima Steud.

The discovery and deployment of new broad-spectrum resistance (R) genes from cultivated rice and its wild relatives is a strategy to broaden the genetic basis of modern rice cultivars to combat rice blast disease. Oryza glaberrima possessing many valuable traits for tolerance to biotic and abiotic stresses, is an elite gene pool for improvement of Asian cultivated rice. An introgression line IL106 derived from O. glaberrima (Acc. IRGC100137) confers complete resistance to Magnaporthe oryzae in blast nursery. Genetic analysis using 2185 BC6F2 progenies derived from a cross between IL106 and the recurrent parent Dianjingyou 1 showed that IL106 harbors a single dominant resistance gene against M. oryzae strain 09BSH-10-5A. This gene was preliminarily mapped on the long arm of chromosome 6 of rice in a region of ca. 0.9 cM delimited by two SSR markers (RM20650 and RM20701). In order to finely map this gene, 17,100 additional progenies were further analyzed. As a result, this gene was further narrowed down to a region flanked by two molecular markers STS69-15 and STS69-7, and co-segregated with 3 molecular markers, RM20676, STS69-21 and STS69-22 on the long arm of chromosome 6. Based on reference genome sequences, this R gene was mapped in silico in 76.1-Kb and 67.7-Kb physical intervals, and containing 4 and 3 NBS-LRR candidate genes in O. sativa cultivar Nipponbare and O. glaberrima cultivar CG14, respectively. Because no blast resistance gene was finely mapped in this physical interval before, this R gene was considered as not described yet and designated as Pi69(t), which is the first identified and finely mapped blast R gene from O. glaberrima, as far as we know. Evaluation of IL106 with 151 blast strains collected from 6 countries in Asia showed that 148 strains are avirulent on IL106, suggesting that Pi69(t) is a broad-spectrum blast R gene, and a promising resistant resource for improvement of Asian cultivated rice.

Novel haplotypes and networks of AVR-Pik alleles in Magnaporthe oryzae.

BACKGROUND: Rice blast disease is one of the most destructive fungal disease of rice worldwide. The avirulence (AVR) genes of Magnaporthe oryzae are recognized by the cognate resistance (R) genes of rice and trigger race-specific resistance. The variation in AVR is one of the major drivers of new races. Detecting the variation in the AVR gene in isolates from a population of Magnaporthe oryzae collected from rice production fields will aid in evaluating the effectiveness of R genes in rice production areas. The Pik gene contains 5 R alleles (Pik, Pikh, Pikp, Pikm and Piks) corresponding to the AVR alleles (AVR-Pik/kh/kp/km/ks) of M. oryzae. The Pik gene specifically recognizes and prevents infections by isolates of M. oryzae that contain AVR-Pik. The molecular variation in AVR-Pik alleles of M. oryzae and Pik alleles of rice remains unclear. RESULTS: We studied the possible evolutionary pathways of AVR-Pik alleles by analyzing their DNA sequence variation and assaying their avirulence to the cognate Pik alleles of resistance genes under field conditions in China. The results of PCR products from genomic DNA showed that 278 of the 366 isolates of M. oryzae collected from Yunnan Province, China, carried AVR-Pik alleles. Among the isolates from six regions of Yunnan, 66.7-90.3% carried AVR-Pik alleles. Moreover, 10 AVR-Pik haplotypes encoding five novel AVR-Pik variants were identified among 201 isolates. The AVR-Pik alleles evolved to virulent from avirulent forms via stepwise base substitution. These findings demonstrate that AVR-Pik alleles are under positive selection and that mutations are responsible for defeating race-specific resistant Pik alleles in nature. CONCLUSIONS: We demonstrated the polymorphism and distribution of AVR-Pik alleles in Yunnan Province, China. By pathogenicity assays used to detect the function of the different haplotypes of AVR-Pik, for the first time, we showed the avoidance and stepwise evolution of AVR-Pik alleles in rice production areas of Yunnan. The functional AVR-Pik possesses diversified sequence structures and is under positive selection in nature.

Wheat Stripe Rust Epidemic and Virulence of Puccinia striiformis f. sp. tritici in China in 2002.

In China, wheat stripe rust, caused by Puccinia striiformis f. sp. tritici, is one of the most destructive diseases of wheat and can cause severe yield losses when susceptible cultivars are grown and weather conditions are favorable for the disease. Wheat stripe rust most frequently affects the winter wheat growing areas in Northwest, Southwest, and North China, and the spring wheat growing areas in Northwest China. In the 2001-2002 growing season, a widespread stripe rust epidemic affected about 6.6 million hectares of wheat in 11 provinces: Si-chuan, Chongqing, eastern Gansu, southern and western Shaanxi, southern and central Ningxia, Yunnan, Guizhou, Hubei, Henan, southern and central Hebei, and Shandong. The epidemic could be attributed to relatively warm weather from November 2001 to March 2002, high frequencies of stripe rust races CYR31 and CYR32, and widely grown susceptible cultivars. Race CYR31 was virulent on the Chinese differential cultivars Trigo Eureka, Fulhard, Lutescens 128, Mentana, Virgilio, Abbondanza, Early Premium, Funo, Danish 1, Fengchan 3, Lovrin 13, Shui-yuan 11, Lovrin 10, and Hybrid 46. Race CYR32 had all the virulence factors of CYR31, plus virulences on Chinese differential cultivars Jubilejina 2 and Kangyin 655, i.e., CYR32 was virulent on all differential cultivars, except Zhong 4. When tested on the world and European differential and some other resistant genotypes, CYR32 was virulent on Chinese 166 (Yr1), Heines VII (Yr2, Yr25, and YrHVII), Vilmorin 23 (Yr3a and Yr4a), Heines Kolben (Yr6 and YrHK), Lee (Yr7, Yr22, and Yr23), Clement (Yr9, Yr25, YrCle), VPM1 (Yr17), Selkirk (Yr27), Anza (YrA), Carstens V (YrCV1, YrCV2, and YrCV3), Gaby (YrG), Strubes Dickkopf (Yr25), and Suwon 92/Omar (YrSO). Resistance genes in Triticum spelta album (Yr5), Zhong 4, and Moro (Yr10 and YrMor) were effective against all races identified.