Ryan Parks

Virulence Structure of the Eastern U.S. Wheat Powdery Mildew Population

Little is known about the population structure of wheat powdery mildew in the eastern United States, and the most recent report on virulence in this population involved isolates collected in 1993-94. In the present study, wheat leaves naturally infected with powdery mildew were collected from 10 locations in the southeastern United States in 2003 and 2005 and a collection of 207 isolates was derived from single ascospores. Frequencies of virulence to 16 mildew resistance (Pm) genes were determined by inoculating the isolates individually on replicated plates of detached leaves of differential wheat lines. These virulence frequencies were used to infer local effectiveness of Pm genes, estimate virulence complexity, detect significant associations between pairs of pathogen avirulence loci, and assess whether phenotypic differences between pathogen subpopulations increased with geographic distance. In both years, virulence to Pm3a, Pm3c, Pm5a, and Pm7 was present in more than 90% of sampled isolates and virulence to Pm1a, Pm16, Pm17, and Pm25 was present in fewer than 10% of isolates. In each year, 71 to 88% of all sampled isolates possessed one of a few multilocus virulence phenotypes, although there were significant differences among locations in frequencies of virulence to individual Pm genes. Several significant associations were detected between alleles for avirulence to pairs of Pm genes. Genetic (phenotypic) distance between isolate subpopulations increased significantly (R2 = 0.40, P < 0.001) with increasing geographic separation; possible explanations include different commercial deployment of Pm genes and restricted gene flow in the pathogen population.

Molecular characterization of a new powdery mildew resistance gene Pm54 in soft red winter wheat

Key messageA new powdery mildew resistance genePm54was identified on chromosome 6BL in soft red winter wheat.AbstractPowdery mildew is causing increasing damage to wheat production in the southeastern USA. To combat the disease, a continuing need exists to discover new genes for powdery mildew resistance and to incorporate those genes into breeding programs. Pioneer® variety 26R61 (shortened as 26R61) and AGS 2000 have been used as checks in the Uniform Southern Soft Red Winter Wheat Nursery for a decade, and both have provided good resistance across regions during that time. In the present study, a genetic analysis of mildew resistance was conducted on a RIL population developed from a cross of 26R61 and AGS 2000. Phenotypic evaluation was conducted in the field at Plains, GA, and Raleigh, NC, in 2012 and 2013, a total of four environments. Three quantitative trait loci (QTL) with major effect were consistently detected on wheat chromosomes 2BL, 4A and 6BL. The 2BL QTL contributed by 26R61 was different from Pm6, a widely used gene in the southeastern USA. The other two QTL were identified from AGS 2000. The 6BL QTL was subsequently characterized as a simple Mendelian factor when the population was inoculated with a single Blumeria graminis f. sp. tritici (Bgt) isolate in controlled environments. Since there is no known powdery mildew resistance gene (Pm) on this particular location of common wheat, the gene was designated Pm54. The closely linked marker Xbarc134 was highly polymorphic in a set of mildew differentials, indicating that the marker should be useful for pyramiding Pm54 with other Pm genes by marker-assisted selection.

Rye-derived powdery mildew resistance gene Pm8 in wheat is suppressed by the Pm3 locus

Genetic suppression of disease resistance is occasionally observed in hexaploid wheat or in its interspecific crosses. The phenotypic effects of genes moved to wheat from relatives with lower ploidy are often smaller than in the original sources, suggesting the presence of modifiers or partial inhibitors in wheat, especially dilution effects caused by possible variation at orthologous loci. However, there is little current understanding of the underlying genetics of suppression. The discovery of suppression in some wheat genotypes of the cereal rye chromosome 1RS-derived gene Pm8 for powdery mildew resistance offered an opportunity for analysis. A single gene for suppression was identified at or near the closely linked storage protein genes Gli-A1 and Glu-A3, which are also closely associated with the Pm3 locus on chromosome 1AS. The Pm3 locus is a complex of expressed alleles and pseudogenes embedded among Glu-A3 repeats. In the current report, we explain why earlier work indicated that the mildew suppressor was closely associated with specific Gli-A1 and Glu-A3 alleles, and predict that suppression of Pm8 involves translated gene products from the Pm3 locus.

Appearance of Powdery Mildew of Wheat Caused by Blumeria graminis f. sp. tritici on Pm17-Bearing Cultivars in North Carolina

Pm17 is a gene for resistance to powdery mildew caused by Blumeria graminis (DC.) E.O. Speer f. sp. tritici. The gene was first confirmed in the wheat-rye translocation cultivar Amigo (1). In Amigo, the translocation is T1AL-1RS and the 1RS arm has the gene Pm17. In the mid-Atlantic United States, at least two widely deployed soft red winter wheat (Triticum aestivum L.) cultivars, McCormick (2) and Tribute (3), possess Pm17 inherited from Amigo. Before 2009, low frequencies of mostly intermediate virulence to Pm17 were detected among isolates from research plots of highly susceptible cultivars (4), but Pm17-bearing cultivars remained immune to mildew in the field. In April 2009, moderately severe powdery mildew was observed for the first time throughout plots of McCormick, Tribute, and other cultivars in both Kinston and Raleigh, NC. At Kinston, Pm17 virulence was observed at two research sites, separated by approximately 10 km, throughout plots of Amigo, McCormick, Tribute, and the hard red winter wheat cultivar TAM 303, which also contains Pm17. In the same month, virulence to Pm17 was observed in Raleigh throughout rows and plots of Amigo and TAM 303. In Kinston and Raleigh, ratings of powdery mildew severity on the Pm17-containing cultivars were 4 or 5 on a scale of 0 to 9, with 0 being the absence of mildew pustules and 9 the most severe mildew infection. Mildew was observed on leaves of all ages. Mildewed leaves were collected from field plots of all four Pm17-bearing cultivars, and an assay to confirm Pm17 virulence was conducted in the laboratory. Mixed-isolate cultures were derived from the leaves and a detached-leaf assay was performed using Amigo, which is the standard Pm17 differential (4). All tested cultures were fully to moderately virulent on Pm17 and all were fully virulent on the susceptible control Chancellor. In the field, chasmothecia (sexual fruiting bodies) were observed on Pm17-bearing cultivars. Together with the quantitatively varying Pm17 virulence detected in the laboratory assay, this suggests that multiple strains of Pm17-virulent B. graminis f. sp. tritici may be present in the field, although that has not yet been demonstrated. Pm17 has protected wheat from powdery mildew over a substantial area in the mid-Atlantic United States. The loss of Pm17 is the most important virulence shift in the U.S. wheat powdery mildew population since Pm4a became ineffective around 2002. Isolates virulent to Pm17 can be expected to appear and multiply in wheat-producing states of the mid-Atlantic United States, including Delaware, Maryland, Virginia, North Carolina, South Carolina, and Georgia. Thus, the urgency of developing and releasing wheat cultivars with other sources of effective mildew resistance is heightened. References: (1) B. Friebe et al. Euphytica 91:59, 1996. (2) C. A. Griffey et al. Crop Sci. 45:416, 2005. (3) C. A. Griffey et al. Crop Sci. 45:419, 2005. (4) R. Parks et al. Plant Dis. 92:1074, 2008.

Differentiation Among Blumeria graminis f. sp. tritici Isolates Originating from Wild Versus Domesticated Triticum Species in Israel.

Israel and its vicinity constitute a center of diversity of domesticated wheat species (Triticum aestivum and T. durum) and their sympatrically growing wild relatives, including wild emmer wheat (T. dicoccoides). We investigated differentiation within the forma specialis of their obligate powdery mildew pathogen, Blumeria graminis f. sp. tritici. A total of 61 B. graminis f. sp. tritici isolates were collected from the three host species in four geographic regions of Israel. Genetic relatedness of the isolates was characterized using both virulence patterns on 38 wheat lines (including 21 resistance gene differentials) and presumptively neutral molecular markers (simple-sequence repeats and single-nucleotide polymorphisms). All isolates were virulent on at least some genotypes of all three wheat species tested. All assays divided the B. graminis f. sp. tritici collection into two distinct groups, those from domesticated hosts and those from wild emmer wheat. One-way migration was detected from the domestic wheat B. graminis f. sp. tritici population to the wild emmer B. graminis f. sp. tritici population at a rate of five to six migrants per generation. This gene flow may help explain the overlap between the distinct domestic and wild B. graminis f. sp. tritici groups. Overall, B. graminis f. sp. tritici is significantly differentiated into wild-emmer and domesticated-wheat populations, although the results do not support the existence of a separate f. sp. dicocci.

Structure and Migration in U.S. Blumeria graminis f. sp. tritici Populations.

While wheat powdery mildew occurs throughout the south-central and eastern United States, epidemics are especially damaging in the Mid-Atlantic states. The structure of the U.S. Blumeria graminis f. sp. tritici population was assessed based on a sample of 238 single-spored isolates. The isolates were collected from 16 locations in 12 states (18 site-years) as chasmothecial samples in 2003 or 2005, or as conidial samples in 2007 or 2010. DNA was evaluated using nine single nucleotide polymorphism (SNP) markers in four housekeeping genes, and 10 simple sequence repeat (SSR) markers. The SSR markers were variably polymorphic, with allele numbers ranging from 3 to 39 per locus. Genotypic diversity was high (210 haplotypes) and in eight of the site-years, every isolate had a different SSR genotype. SNP haplotypic diversity was lower; although 15 haplotypes were identified, the majority of isolates possessed one of two haplotypes. The chasmothecial samples showed no evidence of linkage disequilibrium (P = 0.36), while the conidial samples did (P = 0.001), but the two groups had nearly identical mean levels of genetic diversity, which was moderate. There was a weakly positive relationship between genetic distance and geographic distance (R(2) = 0.25, P = 0.001), indicating modest isolation by distance. Most locations in the Mid-Atlantic and Great Lakes regions clustered together genetically, while Southeast locations formed a distinct but adjacent cluster; all of these were genetically separated from Southern Plains locations and an intermediate location in Kentucky. One-way migration was detected at a rate of approximately five individuals per generation from populations west of the Appalachian Mountains to those to the east, despite the fact that the Atlantic states experience more frequent and damaging wheat mildew epidemics. Overall, the evidence argues for a large-scale mosaic of overlapping populations that re-establish themselves from local sources, rather than continental-scale extinction and re-establishment, and a low rate of long-distance dispersal roughly from west to east, consistent with prevailing wind directions.