Project description:Barley (Hordeum vulgare L.) is amongst the oldest domesticated crop plants and remains one of the world's most important crop species. It is diploid with a haploid genome of 5.1 gigabases (Gb), twice the size of those of human and maize, and closely related to the most widely grown crop, hexaploid wheat. To meet global demand for food, feed and fibre, it is commonly agreed that reference genome sequences of our crop plants are urgently required to enable genome-assisted crop improvement. As part of the The International Barley Genome Sequencing Consortium (IBSC) we present here raw data obtained from Illumina (GAII) sequencing of RNA samples from nine different barley cultivars for the purpose of Single Nucleotide Polymorphism (SNP) discovery.
Project description:RNA was isolated from 23 old barley plants (shoots and roots), line Rolap. PARE libraries were constructed for both barley organs, followed by sequencing of NGS libraries.
Project description:We provide raw gene sequences of 174 flowering time regulatory genes and gene othologs across a large barley population (895 barley lines) selected from a collection of landrace, cultivated barley, and research varieties of diverse origin. This set represents the whole variety of cultivated barley lifeforms, namely two- and six-row genotypes with winter, spring, and facultative growth habits. We applied a target capture method based on in-solution hybridization using the myBaits® technology (Arbor Biosciences, Ann Arbour, MI, USA) which is based on in-solution biotinylated RNA probes. Baits were designed for flowering time regulatory genes and gene othologs, and used for production of 80mer capture oligonucleotides for hybridization. Genomic DNA was extracted from leaves of a single two-week old barley plant per variety using the cetyl-trimethyl-ammonium bromide (CTAB) method. Physical shearing of genomic DNA was performed with an average size of 275 bp. Library preparation was conducted with KAPA Hyper Prep Kit (KAPA Biosystems, Wilmington, MA). Hybridization of customised RNA baits with capture pools was performed at 65°C for 24 hours. Each pooled sequence capture library was sequenced on an Illumina HiSeq3000 instrument using three lanes to generate paired-end reads per sample. Genome sequencing was conducted at AgriBio, (Centre for AgriBioscience, Bundoora, VIC, Australia).
Project description:To provide comprehensive spatiotemporal information about biological processes in developing grains of cultivated barley (Hordeum vulgare subsp. vulgare), we performed a chromatin immunoprecipitation of H3K27me3 followed by high-throughput sequencing (ChIP-seq) in barley endosperm at 16 days after pollination.
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:We used a transcriptome sequencing approach to analyze different expression levels of three barley varieties under both infected and uninfected conditions
Project description:Transcript levels of barley genes were examined in the wheat-barley chromosome addition lines having one of six barley chromomes, 2H, 3H, 4H, 5H, 6H and 7H. ****[PLEXdb(http://www.plexdb.org) has submitted this series at GEO on behalf of the original contributor, Seungho Cho. The equivalent experiment is BB8 at PLEXdb.]
Project description:RNA was isolated from 23 old barley plants (shoots and roots), line Rolap. We used a modified method that allows for enrichment of small RNAs. Libraries were prepared using TruSeq Small RNA Library Preparation Kit (Illumina), followed by sequencing of NGS libraries.