Project description:Wheat grain storage proteins (GSPs) make up most of the protein content of grain and determine flour end-use value. The synthesis and accumulation of GSPs depend highly on nitrogen (N) and sulfur (S) availability and it is important to understand the underlying control mechanisms. Here we studied how the einkorn (Triticum monococcum ssp. monococcum) grain proteome responds to different amounts of N and S supply during grain development. Two subproteomes were analyzed : albumin-globulin and nuclear proteins.
Project description:Purpose: The goal of this study is to compare endothelial small RNA transcriptome to identify the target of OASL under basal or stimulated conditions by utilizing miRNA-seq. Methods: Endothelial miRNA profilies of siCTL or siOASL transfected HUVECs were generated by illumina sequencing method, in duplicate. After sequencing, the raw sequence reads are filtered based on quality. The adapter sequences are also trimmed off the raw sequence reads. rRNA removed reads are sequentially aligned to reference genome (GRCh38) and miRNA prediction is performed by miRDeep2. Results: We identified known miRNA in species (miRDeep2) in the HUVECs transfected with siCTL or siOASL. The expression profile of mature miRNA is used to analyze differentially expressed miRNA(DE miRNA). Conclusions: Our study represents the first analysis of endothelial miRNA profiles affected by OASL knockdown with biologic replicates.
Project description:Amylase/trypsin-inhibitors (ATIs) are putative triggers of nonceliac gluten sensitivity, but contents of ATIs in different wheat species were not available. Therefore, the predominant ATIs 0.19 + 0.53, 0.28, CM2, CM3, and CM16 in eight cultivars each of common wheat, durum wheat, spelt, emmer, and einkorn grown under the same environmental conditions were quantitated by targeted liquid chromatography-tandem mass spectrometry (LC−MS/MS) and stable isotope dilution assays using specific marker peptides as internal standards. The results were compared to a label-free untargeted LC−MS/MS analysis, in which protein concentrations were determined by intensity based absolute quantitation. Both approaches yielded similar results. Spelt and emmer had higher ATI contents than common wheat, with durum wheat in between. Only three of eight einkorn cultivars contained ATIs in very low concentrations. The distribution of ATI types was characteristic for hexaploid, tetraploid, and diploid wheat species and suitable as species-specific fingerprint. The results point to a better tolerability of einkorn for NCGS patients, because of very low total ATI contents.
Project description:A cDNA library was constructed by Novogene (CA, USA) using a Small RNA Sample Pre Kit, and Illumina sequencing was conducted according to company workflow, using 20 million reads. Raw data were filtered for quality as determined by reads with a quality score > 5, reads containing N < 10%, no 5' primer contaminants, and reads with a 3' primer and insert tag. The 3' primer sequence was trimmed and reads with a poly A/T/G/C were removed
Project description:In wheat, the main staple crop in many regions of the word, one major challenge is to improve the yield potential while maintaining the grain quality, which is mainly defined by its protein concentration and composition. To achieve this goal, it is essential to considerate the nitrogen (N) and sulfur (S) nutrition, which strongly influences the grain storage protein (GSP) composition. Nowadays, the limitation of nitrogen inputs and the sulfur deficiency recently observed in soils represent major difficulties to control the grain quality. Thus, identification of the molecular mechanisms that control the accumulation of GSP in response to N and S supply is necessary. In this study, we applied four post-anthesis N and S treatments to an einkorn (Triticum monococcum) culture. Effects of the treatments on the grain transcriptome and metabolome were evaluated at different times during grain development. From the 386 differentially expressed genes and the 21 metabolites differentially accumulated, our results have revealed a strong impact of a high-N supply without any added S. A large majority of genes were transiently influenced by the treatments while others showed strong modifications of their kinetics of expression during grain filling. We hypothesized the role of several genes (e.ge.g. sulfate transporters, transcriptional regulators) in the adjustment of the N-to-S ratio in response to a S deficiency. These genes could coordinate the amino acid pool necessary for GSP synthesis. These new results contribute in facing the challenge of maintaining wheat grain quality for the development of more sustainable agriculture.
Project description:An untargeted liquid chromatography tandem mass spectrometry method was used to analyze the content of peptides with celiac disease (CD) active epitope in the five wheat species common wheat, spelt, durum wheat, emmer and einkorn. In total, 494 peptides with CD active epitope were identified. Relevant differences between the species concerning their CD immunoractive potential based on the distribution of CD-active epitopes and relative quantities of peptides with CD-active epitope were demonstrated.
