BackgroundResistance in plants to pathogen attack can be qualitative or quantitative. For the latter, hundreds of quantitative trait loci (QTLs) have been identified, but the mechanisms of resistance are largely unknown. Integrated non-target metabolomics and proteomics, using high resolution hybrid mass spectrometry, were applied to identify the mechanisms of resistance governed by the fusarium head blight resistance locus, Fhb1, in the near isogenic lines derived from wheat genotype Nyubai.FindingsThe metabolomic and proteomic profiles were compared between the near isogenic lines (NIL) with resistant and susceptible alleles of Fhb1 upon F. graminearum or mock-inoculation. The resistance-related metabolites and proteins identified were mapped to metabolic pathways. Metabolites of the shunt phenylpropanoid pathway such as hydroxycinnamic acid amides, phenolic glucosides and flavonoids were induced only in the resistant NIL, or induced at higher abundances in resistant than in susceptible NIL, following pathogen inoculation. The identities of these metabolites were confirmed, with fragmentation patterns, using the high resolution LC-LTQ-Orbitrap. Concurrently, the enzymes of phenylpropanoid biosynthesis such as cinnamyl alcohol dehydrogenase, caffeoyl-CoA O-methyltransferase, caffeic acid O-methyltransferase, flavonoid O-methyltransferase, agmatine coumaroyltransferase and peroxidase were also up-regulated. Increased cell wall thickening due to deposition of hydroxycinnamic acid amides and flavonoids was confirmed by histo-chemical localization of the metabolites using confocal microscopy.ConclusionThe present study demonstrates that the resistance in Fhb1 derived from the wheat genotype Nyubai is mainly associated with cell wall thickening due to deposition of hydroxycinnamic acid amides, phenolic glucosides and flavonoids, but not with the conversion of deoxynivalenol to less toxic deoxynivalenol 3-O-glucoside.
Marker-assisted breeding provides an opportunity for wheat breeders to introgress/pyramid genes of interest into breeding lines and to identify genes and/or quantitative trait loci in germplasm to be used as parents. Molecular markers were deployed to assist selection for disease resistance, agronomic and quality traits in several wheat cultivars released for commercial cultivation in Canada. Marker-assisted breeding is routinely used in most wheat breeding programmes for rust resistance (leaf, stem and stripe rust), orange wheat blossom midge resistance, high grain protein concentration, Fusarium head blight and common bunt resistance. Markers are being used selectively within breeding programmes to target traits that relate to market class or regional adaptation. For example, marker-assisted breeding for low lipoxygenase activity and low grain cadmium is being performed in durum breeding programmes and for enhancing stem solidness in programmes targeting resistance to the wheat stem sawfly. Markers are also being utilized for ergot resistance in durum wheat. Increased gluten strength is being selected with a marker for the overexpression of the Bx7 high-molecular-weight glutenin subunit. Marker-assisted breeding is also being used to pyramid resistance genes against a group of stem rust races related to TTKS (Ug99), a disease that poses a serious threat to global wheat production. Development of tightly linked diagnostic markers and high-throughput genotyping with SNP markers will result in more effective molecular wheat breeding in the near future and will open the door to genomic selection.
Bread wheat and durum wheat were examined for linkage disequilibrium (LD) using microsatellite markers distributed across the genome. The allele database consisted of 189 bread wheat accessions genotyped at 370 loci and 93 durum wheat accessions genotyped at 245 loci. A significance level of p< 0.001 was set for all comparisons. The bread and durum wheat collections showed that 47.9% and 14.0% of all locus pairs were in LD, respectively. LD was more prevalent between loci on the same chromosome compared with loci on independent chromosomes and was highest between adjacent loci. Only a small fraction (bread wheat, 0.9%; durum wheat, 3.2%) of the locus pairs in LD showed R2 values > 0.2. The LD between adjacent locus pairs extended (R2 > 0.2) approximately 2-3 cM, on average, but some regions of the bread and durum wheat genomes showed high levels of LD (R2 = 0.7 and 1.0, respectively) extending 41.2 and 25.5 cM, respectively. The wheat collections were clustered by similarity into subpopulations using unlinked microsatellite data and the software Structure. Analysis within subpopulations showed 14- to 16-fold fewer locus pairs in LD, higher R2 values for those pairs in LD, and LD extending further along the chromosome. The data suggest that LD mapping of wheat can be performed with simple sequence repeats to a resolution of <5 cM.
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