Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Investigation of barley brittle stem mutants

ABSTRACT: The barley brittle stem mutants, fs2, designated X054 and M245, have reduced levels of cellulose compared with their isogenic parents Ohichi and Shiroseto. A custom-designed microarray, based on Agilent technology and including genes involved in cell wall metabolism, was used to compare transcript levels in the mutant and parental lines. For both mutants, the microarray revealed a marked decrease in mRNA for the HvCesA4 cellulose synthase gene in specific zones of stem internodes, and this was confirmed by quantitative PCR. Genes expression was measured for the upper and lower zones from the 4th internodes of stems. Plant were at Zadocks developmental stage 49. Gene expression was compared between mutants and their wildtype parents.

ORGANISM(S): Hordeum vulgare

SUBMITTER: Ute Baumann

PROVIDER: E-GEOD-17873 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

ACCESS DATA

Publications

A customized gene expression microarray reveals that the brittle stem phenotype fs2 of barley is attributable to a retroelement in the HvCesA4 cellulose synthase gene.

Burton Rachel A RA Ma Gang G Baumann Ute U Harvey Andrew J AJ Shirley Neil J NJ Taylor Jillian J Pettolino Filomena F Bacic Antony A Beatty Mary M Simmons Carl R CR Dhugga Kanwarpal S KS Rafalski J Antoni JA Tingey Scott V SV Fincher Geoffrey B GB

Plant physiology 20100607 4

The barley (Hordeum vulgare) brittle stem mutants, fs2, designated X054 and M245, have reduced levels of crystalline cellulose compared with their parental lines Ohichi and Shiroseto. A custom-designed microarray, based on long oligonucleotide technology and including genes involved in cell wall metabolism, revealed that transcript levels of very few genes were altered in the elongation zone of stem internodes, but these included a marked decrease in mRNA for the HvCesA4 cellulose synthase gene ...[more]

PMID: 20530215

Similar Datasets

Project description:Transcriptional profiling of hop samples throughout in vitro culture on medium with sucrose and hormones IAA and BAP Study of gene expression involved in stress response during in vitro culture and in organogenic nodule formation Hop (Humulus lupulus L.) is an economically important plant forming organogenic nodules which can be used for genetic transformation and micropropagation. We are interested in the mechanisms underlying reprogramming of cells through stress and hormone treatments. To investigate the spatio-temporal sequence of events that underlies competence acquisition for organogenesis three main stages were selected: (i) internodes at the time of excision from the parent plant (T0); (ii) internodes grown for 15 days on culture medium in which several prenodular structures are formed inside the calli (T15d); and (iii) nodule forming tissue after 28 days of culture (T28d). Additionally a control was included corresponding to 28 days of culture without hormones, lacking nodule formation and plantlet regeneration abilities, in order to identify genes specifically involved in morphogenesis (T28dWH). Organogenic nodule formation is a morphogenic process that shares features with somatic embryogenesis though in the former no shoot/root pole is established and plantlet regeneration can occur from different peripheral regions of nodules. Previous studies have shown that the organogenesis-determining period (when cells are determined to form nodules) occurs in between 15 and 25 days of culture corresponding to prenodular and first nodular stages. When nodules are fully developed they are surrounded by layers of elongated, highly vacuolated cells that may degenerate when nodules start differentiating plantlets. Plantlet regeneration from organogenic nodules can be observed after 45 days of culture. However, organogenic nodules after 28 days of culture are already determined to undergo plantlet regeneration, since they no longer require exogenous supply of hormones. Explants cultured in medium without hormones can develop incipient prenodular structures with vascular tissue but not nodules, thus lacking morphogenic potential. Keywords: Plant development, Time course 3 time points: 3 Biological replicates and 1 technical replicate (1 dye-swap). 1 control at beginning of culture and 1 control wihout hormones. One replicate per array. Each clone printed twice in the same sub-grid

Project description:Alfalfa, [Medicago sativa (L.) sativa], a widely-grown perennial forage has potential for development as a cellulosic ethanol feedstock. The application of genomic approaches would advance development of alfalfa as a cellulosic feedstock. However, the genomics of alfalfa, a non-model species, is still in its infancy. The recent advent of RNA-Seq, a massively parallel sequencing method for transcriptome analysis, provides an opportunity to expand the identification of alfalfa genes and polymorphisms, and conduct in-depth transcript profiling. Cell walls in stems of alfalfa genotype 708 have higher cellulose and lower lignin concentrations compared to cell walls in stems of genotype 773. Using the Illumina GA-II platform, a total of 198,861,304 expression sequence tags (ESTs, 76 bp in length) were generated from cDNA libraries derived from elongating stem (ES) and post-elongation stem (PES) internodes of 708 and 773. These ESTs were de novo assembled into 132,153 unique sequences. By combining the de novo assembled ESTs (132,153 sequences) with our previously identified EST sequences (341,984 sequences, unpublished data), and the ESTs available from GenBank (12,371 sequences), we built the first Alfalfa Gene Index (MSGI 1.0). MSGI 1.0 contains 124,025 unique sequences including 22,729 tentative consensus sequences (TCs), 22,315 singletons and 78,981 pseudo-singletons. We identified a total of 1, 294 simple sequence repeats (SSR) among the sequences in MSGI 1.0. In addition, a total of 10,826 single nucleotide polymorphisms (SNPs) were predicted between the two genotypes. Transcript profiling of stem internodes of genotypes 708 and 773 was conducted by quantifying the number of Illumina EST reads that were mapped to sequences in MSGI 1.0. We identified numerous candidate genes that may play a role in stem development as well as candidate genes that may contribute to the differences in cell wall composition in stems of the two genotypes. Our results demonstrate that RNA-Seq can be successfully used for gene identification, polymorphism detection and transcript profiling in alfalfa, a non-model, allogamous, autotetraploid species. The alfalfa gene index (MSGI 1.0) assembled in this study, and the SNPs, SSRs and candidate genes identified can be used to improve alfalfa as a cellulosic feedstock. Examination of 2 different tissue types at different developmental stages (Elongating vs. post-elongation stem internodes) in two alfalfa genotypes (708 and 773) with divergent cell wall composition in stems.

Dataset Information

Investigation of barley brittle stem mutants

Publications

A customized gene expression microarray reveals that the brittle stem phenotype fs2 of barley is attributable to a retroelement in the HvCesA4 cellulose synthase gene.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets