Project description:Transcriptome analysis of Barley flag leaf at several stages of senescence. The experiment includes transcriptomic data of the cv.Karl and its early-senescing near-isogenic line '10_11'(jukanti et al.2008)
Project description:The assays performed are aimed at reflecting the immense physiological changes taking place in a senescing barley flag leaf, compared to a non-senescing young flag.
Project description:This study was aimed at deciphering the impact of drought and heat on genome-wide gene expression in flag leaf of barley. We employed high-throughput sequencing of mRNA to identify genes that are associated with response to drought or heat and to their combination. Our study demonstrated that under combined stress, drought was the dominant factor affecting genes expression. It was also confirmed for phenotypic traits and chlorophyll fluorescence parameters. Drought- and heat-responsive genes were associated majorly with photosynthesis, abscisic acid signaling and lipids transport. Dehydrin encoding genes were found to be universal stress-responsive genes. Stress-induced genes specific to the flag leaf size were also found. This research provided novel insight into molecular mechanisms of barley flag leaf that determine drought and heat response, also during their co-occurrence.
Project description:Barley is an important cereal crop all over the world. Detailed molecular characterization of barley provides the basis for development of improved cultivars, stress and drought-resistant plants. We here present an LC-MS/MS-based proteomics study of barley leaf aimed at optimization of methods to achieve efficient and unbiased trypsin digestion of proteins prior to LC-MS/MS based sequencing and quantification of peptides. We evaluated two spin filter-aided sample preparation protocols using either sodium dodecyl-sulphate (SDS) or sodium deoxycholate (SDC), and three in-solution digestion (ISD) protocols using SDC or trichloroacetic acid/acetone precipitation. The proteomics workflow identified up to 1800 barley proteins based on sequencing of up to 7700 peptides per sample. The two spin filter-based protocols provided a 17-38% higher efficiency than the ISD protocols, including more proteins of low abundance. Among the ISD protocols, a simple one-step reduction and S-alkylation method (OP-ISD) was the most efficient for barley leaf sample preparation; it identified and quantified 1500 proteins and displayed higher peptide-to-protein inference ratio and higher average amino acid sequence coverage of proteins. The two spin filter-aided sample preparation protocols are compatible with TMT labeling for quantitative proteomics studies. They exhibited complementary performance as about 30% of the proteins were identified by either one or the other protocol, but also demonstrated a positive bias for membrane proteins when using SDC as detergent. We provide detailed protocols for efficient barley protein sample preparation for LC-MS/MS-based proteomics studies. Spin filter-based protocols are the most efficient for the preparation of barley leaf samples for MS-based proteomics, however, a simple protocol provides comparable results although with different peptide digestion profile.
Project description:Plant seeds prepare for germination already during seed maturation. We performed a detailed transcriptome analysis of barley grain maturation, desiccation and germination in two tissue fractions (endosperm/aleurone = e/a and embryo = em) using the Affymetrix barley1 chip. Keywords: time course
Project description:Plant seeds prepare for germination already during seed maturation. We performed a detailed transcriptome analysis of barley grain maturation, desiccation and germination in two tissue fractions (endosperm/aleurone = e/a and embryo = em) using the Affymetrix barley1 chip. Experiment Overall Design: Barley developing and germinating seeds were harvested at different time points after flowering (developing) and imbibition (germinating). To further disseect the influence of different tissues, seeds were dissecte and tissues were analyzed individually.
Project description:4plex_barley_2017_01 - transcriptomic assay on embryos after barley seeds gamma_irradiation - Why radiation exposure on seeds at low doses causes stimulation of plant growth after germination? - Barley dry seeds of variety “Nur” were irradiated by gamma-rays (Co-60) at doses 20 Gy (stimulatory) and 100 Gy (inhibitory). Immediately after irradiation we put seeds for germination to Petri dishes containing water filter paper; embryos were taken 2h after irradiation, embryos with embrionic roots were taken 24 and 48h after irradiation. The following comparisons have been made: non-irradiated (0 Gy), 2h vs. 20 Gy, 2h; 0 Gy, 24h vs. 20 Gy, 24h; 0 Gy, 48h vs. 20 Gy, 48h; 0 Gy, 24h vs. 100 Gy, 24h. Three independent biological replicates were used.
Project description:In this study, we focused on a super rice variety, WFYT025, which is widely used in China. Therefore, we took the flag leaves of WFYT025 and its two parents for high-throughput transcriptome sequencing, trying to find some genes related to photosynthesis or transpiration and development of seeds. The aim of our study is (1) we investigated the length, width, and leaf area of flag leaves for WFYT025 and its parents. MPH and HPH were estimated for the heterosis of flag leaf. (2) We selected the flag leaves of hybrid rice WFYT025 and its parents for transcriptome sequencing, the first day and the tenth days after flowering in the environment of early rice and middle rice, respectively. (3) we referred to further investigate the gene regulatory network. Several major sub-networks were found to represents interactions among genes with similar expression profiles via gene regulatory network analysis.