Aegilops tauschii, the diploid wild progenitor of the D subgenome of bread wheat, is a reservoir of genetic diversity for improving bread wheat performance and environmental resilience. Here we sequenced 242 Ae. tauschii accessions and compared them to the wheat D subgenome to characterize genomic diversity. We found that a rare lineage of Ae. tauschii geographically restricted to present-day Georgia contributed to the wheat D subgenome in the independent hybridizations that gave rise to modern bread wheat. Through k-mer-based association mapping, we identified discrete genomic regions with candidate genes for disease and pest resistance and demonstrated their functional transfer into wheat by transgenesis and wide crossing, including the generation of a library of hexaploids incorporating diverse Ae. tauschii genomes. Exploiting the genomic diversity of the Ae. tauschii ancestral diploid genome permits rapid trait discovery and functional genetic validation in a hexaploid background amenable to breeding.

Background Fusarium head blight resistance genes, Fhb1 (for Type-II resistance), Fhb2 (Type-II), and Fhb5 (Type-I plus some Type-II), which originate from Sumai 3, are among the most important that confer resistance in hexaploid wheat. Near-isogenic lines (NILs), in the CDC Alsask (susceptible; n = 32) and CDC Go (moderately susceptible; n = 38) backgrounds, carrying these genes in all possible combinations were developed using flanking microsatellite markers and evaluated for their response to FHB and deoxynivalenol (DON) accumulation in eight environments. NILs were haplotyped with wheat 90 K iSelect assay to elucidate the genomic composition and confirm alleles’ presence. Other than evaluating the effects of three major genes in common genetic background, the study elucidated the epistatic gene interactions as they influence FHB measurements; identified loci other than Fhb1 , Fhb2 , and Fhb5 , in both recurrent and donor parents and examined annotated proteins in gene intervals. Results Genotyping using 81,857 single nucleotide polymorphism (SNP) markers revealed polymorphism on all chromosomes and that the NILs carried < 3% of alleles from the resistant donor. Significant improvement in field resistance (Type-I + Type-II) resulted only among the CDC Alsask NILs, not the CDC Go NILs. The phenotypic response of NILs carrying combinations of Sumai 3 derived genes suggested non-additive responses and Fhb5 was as good as Fhb1 in conferring field resistance in both populations. In addition to Fhb1 , Fhb2 , and Fhb5, four to five resistance improving alleles in both populations were identified and three of five in CDC Go were contributed by the susceptible parent. The introgressed chromosome regions carried genes encoding disease resistance proteins, protein kinases, nucleotide-binding and leucine rich repeats’ domains. Complex epistatic gene-gene interactions among marker loci (including Fhb1 , Fhb2 , Fhb5 ) explained > 20% of the phenotypic variation in FHB measurements. Conclusions Immediate Sumai 3 derivatives carry a number of resistance improving minor effect alleles, other than Fhb1 , Fhb2 , Fhb5 . Results verified that marker-assisted selection is possible for the introgression of exotic FHB resistance genes, however, the genetic background of the recipient line and epistatic interactions can have a strong influence on expression and penetrance of any given gene. Electronic supplementary material The o...

Wheat stripe rust, caused by Puccinia striiformis f. sp. tritici, has been common across Saskatchewan, Canada since 2000. Fifty-nine isolates of P. striiformis f. sp. tritici, the majority of which were collected between 2011 and 2013 from Saskatchewan and southern Alberta, were analyzed for virulence frequency and diversity and compared with isolates characterized in the Pacific Northwest and Great Plains regions of the United States. In all, 31 wheat differentials, including 20 near-isogenic lines and 1 triticale variety, differentiated 59 P. striiformis f. sp. tritici isolates into 33 races, of which one race, C-PST-1, represented 31% of the isolates. None of the races were virulent on Yr5, Yr15, or YrSP. Virulence frequency ranged from 65 to 98% on YrA, Yr2, Yr8, Yr9, Yr27, Yr29, Yr32, YrSu, ‘Heines VII’, and ‘Nord Deprez’. Race C-PST-6 was virulent on the greatest number of the differentials (n = 25) and C-PST-18 on the fewest (n = 14). Discriminant analysis of principal components and multivariate cluster analyses detected three and four major groups, respectively, which differed from each other in terms of virulence spectrum and year of collection. The diversity of the P. striiformis f. sp. tritici population in southern Alberta was greater than in Saskatchewan, which indicated that, although P. striiformis f. sp. tritici is primarily windborne over great distances and does not usually overwinter, there are detectable differences in virulence between these regions of western Canada. Comparative analyses of virulence frequency of Saskatchewan or southern Alberta isolates with isolates representing races from the Great Plains and the Pacific Northwest of the United States indicated greater similarity of Saskatchewan races to the Great Plains despite strong correlations with both parts of the United States. This suggests that the P. striiformis f. sp. tritici population in Saskatchewan is a mixture of inoculum from both parts of the United States.

Wheat stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is an important disease in Canada. The worldwide genetic structure of Pst populations have been characterized, excluding Canada. Here, we elucidated the genetic structure of the western Canadian Pst population using molecular markers, revealing the presence of four divergent lineages with predominantly clonal structure. In the worldwide context, two previously reported lineages were identified: PstS0 (22%), representing an old Northwestern-European and PstS1 (35%), an invasive warm-temperature adapted. Additionally, two new, unreported lineages, PstPr (9%) and PstS1-related (35%), were detected, which produced more telia than other lineages and had double the number of unique recombination events. The PstPr was a recent invasion, and likely evolved in a diverse, recombinant population as it was closely related to the PstS5, PstS7/Warrior, PstS8/Kranich, and PstS9 lineages originating from sexually recombining populations in the centre of diversity. The DNA methylation analysis revealed DNA-methyltransferase1-homologs, providing compelling evidence for epigenetic regulation and as a first report, an average of ∼5%, 5hmC in the Puccinia epigenome merits further investigation. The divergent lineages in the Canadian Pst population with the potential for genetic recombination, as well as epigenetic regulation needs consideration in the context of pathogen adaptation and management.