Project description:A spatial resolved transcriptome for germinating barley grain was obtained for over 14,000 genes at 0, 1, 3 6 and 24 hours after imbibition. The spatial expression of a number of genes was validated using in-situ PCR, quantitative comparison to bulk RNA-seq reveal good correlations, despite the differences in the methodologies, and marker gene definition for tissues/clusters was consistent with previous gene specific studies. Targeted analysis of genes encoding aquaporins, crucial for water and ion movement during germination, revealed specific spatial expression patterns for gene members, and the tissue specific changes in gene members over time. Highly restricted spatial (and temporal) expression patterns were observed for genes encoding cell wall and transcription factors, e.g. one of the strongest expressed transcription factors was a bZIP family member that responds to germinating promoting Karrikin. It’s expression was restricted to the scutellum and the adjacent endosperm (0-3 HAI) and declined afterwards. Discovery-based bio-informatic approaches revealed thousands of genes displayed a spatially restricted pattern within tissues, with gene ontology revealing enriched categories such as auxin related functions of transport and response factors displaying distinct spatial expression patterns in tissues. This provides unprecedented spatial resolved cellular map for germination and specific genes to target for functional genomics to define cellular restricted processes in tissues during germination.
Project description:Analysis of barley grains/seedlings representing six well characterized and distinct germination stages over the course of seed germination and seedling growth.
Project description:Analysis of barley grains/seedlings representing six well characterized and distinct germination stages over the course of seed germination and seedling growth. Three biological replications, six developmental stages.
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:Seeds germination is seriously sensitive to salt stress. The mechanism in response to salt stress during seed germination is still little known. In this study, two genotypes of hulless barley lk621 and lk53 were selected to investigate the molecular mechanism of seeds salinity response during germination stage through RNA-seq and iTRAQ technologies
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:Purpose: The goal of this study was to investigate the mechanisms involved in the initiation and development of crown-roots (CRs) in barley and to estimate the role of cytokinins (CKs) in this process. Method: stranded libraries were obtained from RNA extracted from the stem base of 1 day-after-germination (DAG) and 10DAG-seedlings of wild-type (WT) and AtCKX-overexpressing barley lines (OE-CKX). OE-CKX lines have a reduced content of endogenous CKs and are characterized by a higher number of CRs. Libraries were deep sequenced on Illumina HighSeq platform. Each sample was investigated in three independent biological replicates. Results: Using a data analysis workflow optimized for barley, we identified more than 4000 transcripts differentially expressed in the stem base of 1DAG and 10DAG-seedlings. Expression as determined by RNA-seq was validated by real-time PCR. Our data were compared to the transcriptomic profiling obtained from rice and we were able to identify genes potentially involved in the initiation/development of CRs in barley. Also the use of the transgenic line with altered endogenous CK content allowed us to conclude about the role of CKs in the process. Conclusions: Our study represents the first analysis aiming to understand the initiation and development of CRs in barley.
