Project description:Bread wheat is the most widely cultivated crop worldwide, used in the production of food products and a feed source for animals. Selection tools that can be applied early in the breeding cycle are needed to accelerate genetic gain for increased wheat production while maintaining or improving grain quality if demand from human population growth is to be fulfilled. Proteomics screening assays of wheat flour can assist breeders to select the best performing breeding lines and to filter out the worst performing ones. In this study, we optimised a robust LCMS1 shotgun quantitative proteomics method to screen thousands of wheat genotypes. Using 6 cultivars and 4 replicates, we tested 3 resuspension ratios, 2 extraction buffers, 3 sets of proteases, and multiple LC settings. Protein identifications by LCMS2 was used to select the best parameters. A total 8738 wheat proteins were identified. The best method was validated on an independent set of 96 cultivars and peptides quantities were normalized using sample weights, an internal standard, and Quality Controls. Data mining tools found particularly useful to explore the flour proteome are presented. DOI 10.3390/ijms23020713
Project description:Bread wheat (Triticum aestivum L.) is one of the most valuable cereal crops for the human consumption. Its grain storage proteins greatly impact bread quality, though may cause food intolerances or allergies in susceptible individuals. Consequently, the investigation of a proteome polymorphism among wheat varieties is important to spot the genotypes, which would be promising donors for the breeding of hypoallergenic cereals. Herein, we discovered diversity of grain proteins in three Ukrainian wheat cultivars: ‘Sotnytsia’, ‘Panna’ (both modern selection) and ‘Ukrainka’ (old landrace). Firstly, proteins were isolated with a SDS-containing buffer that allowed extraction of various groups of storage proteins (glutenins, gliadins, globulins and albumins). Secondly, the proteome was profiled by the two-dimensional gel electrophoresis, revealing 810 clearly-separated gel spots. Software-assisted analysis of gel images, showed 66 differentially abundant proteins. Using multi-enzymatic digestion, followed by the tandem mass spectrometry, we identified 49 differentially accumulated proteins. Parallel ultrahigh-performance liquid chromatography profiling and direct mass spectrometry quantification complemented the results. With this approach we quantified 127 proteins, 12 being differentially abundant. Principal component analysis confirmed genotype as a major source of variation in both cases. Non-gluten fraction was the most diverse among investigated bread wheat cultivars. Information from public databases of clinically relevant plant proteins highlighted variable groups of wheat allergens/toxins. Data suggested that one of the modern cultivars contained less health affecting proteins in grain. Finally, we proposed set of genetic landmarks for the development of DNA marker system, which will enable fast and efficient assessment of medical safety of multiple wheat genotypes to facilitate breeding programs.
Project description:The close relationship between diet and health is generally recognized and the growing wellness and consciousness, especially in developed countries, have led to increasing interest for old wheat genotypes, based on perceived health benefits. Although nutritional comparison between old and modern wheat varieties is still controversial, it is generally accepted that old wheat genotypes remained unchanged over the last hundred years. By contrast, modern wheat genotypes are derived by modification of old wheats during the so-called “Green-Revolution” in the second half of the 20th century focusing on obtaining properties in terms of higher grain yield. The present work reports the first comprehensive proteomic profiling and qualitative comparison at the molecular level of metabolic and Chloroform-Methanol (CM)-like protein fractions extracted from mature kernels of two old Sicilian durum wheat landraces, Russello and Timilia Reste Bianche, and Simeto, an improved durum wheat variety widespread in Italy and other Mediterranean countries and chosen as representative of the most widely commercial cultivars. The results obtained reveal that metabolic and CM-like protein fractions of old and modern genotypes present remarkably high similarity with only minor differences. This leads to the conclusion that from a food and nutritional perspective there is a substantial equivalence of the protein composition of the old and modern cultivars.
Project description:In this study, we used dual RNA-sequencing to profile FHB-resistant AC Emerson, FHB-moderately AC Morley, and FHB-susceptible CDC Falcon winter wheat cultivars prior to and in response to Fusarium graminearum at 7 days post inoculation. Differential expression analyses revealed distinct defense responses between resistant and susceptible wheat cultivars including increased mechanical defense through lignin biosynthesis and increased deoxynivalenol (DON) detoxification through UDP-glycosyltransferase activity in resistant cultivars. Further, differential expression analysis in F. graminearum challenging these distinct cultivars revealed changes genes involved in trichothecene mycotoxin biosynthesis.
Project description:Water deficit stress between the booting and grain filling stages significantly affect grain yield and quality of hard red winter wheat. Several stress tolerant cultivars with different adaptation mechanisms have been released and are widely cultivated on the Southern Great Plains of the US. However, the physiological, molecular, and genetic basis of adaptation to drought stress for these cultivars remains unknown. Use of transcriptome profiling to identify drought responsive genes in hexaploid wheat is a challenging process given the quantitative nature of drought stress, genome complexity, and the intricacy of interaction effects. If the information generated from functional genomics studies is to be used in molecular breeding programs for cultivar development, it is highly desirable to use cultivars better adapted for the region. In the current study we used two well-adapted, drought-tolerant, high-yielding, cultivated varieties, TAM 111 and TAM 112, which appear to have different adaptation mechanisms, to identify drought stress induced transcripts during heading and early dough stages. A set of 24 Affymetrix GeneChip wheat genome arrays (2 cultivars; 2 water treatments; 2 sampling stages; 3 biological replicates) from plants subjected to water deficit stress under controlled glasshouse conditions. Differentially expressed genes were identified using a ANOVA (p<0.01) controlling false discovery rate (FDR, q<0.01) using Benjamini Hochberg approach.
Project description:To explore transcriptional regulations in common wheat cultivars Norstar (NO), Manitou (MA), and the near-isogenic lines (NIL) spring Norstar (SN) and winter Manitou (WM) during seasonal cold acclimation.
Project description:Introgression of a high molecular weight glutenin subunit (HMW-GS) gene, 1Ay21*, into commercial wheat cultivars increased overall grain protein content and bread-making quality by unknown mechanisms. As well as increased abundance of 1Ay HMW-GS, 115 differentially expressed proteins (DEPs) were discovered between three cultivars and corresponding introgressed near-isogenic lines (NILs). Functional category analysis showed that the DEPs were predominantly other storage proteins, and proteins involved in protein synthesis, protein folding, protein degradation, stress response and grain development. Nearly half the genes encoding the DEPs showed strong co-expression patterns during grain development. Promoters of these genes are enriched in elements associated with transcription initiation and light response, indicating a potential connection between these cis-elements and grain protein accumulation. A model of how this HMW-GS enhances the abundance of machinery for protein synthesis and maturation during grain filling is proposed. This analysis not only provides insights into how introgression of the 1Ay21* improves grain protein content, but also directs selection of protein candidates for future wheat quality breeding programmes.