Project description:Flowers have a species-specific fertile period during which pollination and fertilization have to occur to initiate seed and fruit development. Within the flower, the functional life span of the ovule containing the female gametophyte is decisive for fertilization and the initiation of seed development. Here we performed an RNA-sequencing based transcriptome analysis of senescing unfertilized ovules during in a time series. We isolated ovules from Arabidopsis thaliana flowers emasculated at stage 12c at three different time points: 2 days after emasculation (DAE), 3 DAE, and 4 DAE. These time points correspond to intact mature ovules (2DAE), early ovule senescence (3 DAE), and late ovule senescence (4 DAE). We extracted total RNA from the ovules in 3 independent biological replicates, thus generating 9 RNA samples in total, for RNA-sequencing by Illumina HiSeq.
Project description:Plant reproduction depends on the concerted activation of many genes to assure the correct communication between pollen and pistil. Here we queried the whole transcriptome of Arabidopsis thaliana in order to identify genes with specific reproductive functions. We used the ATH1 whole genome array to profile wild-type unpollinated pistils and unfertilized ovules in comparison with the expression profile of pistils 0.5, 3.5 and 8.0 hours after pollination KEYWORDS: time course
Project description:Plant reproduction depends on the concerted activation of many genes to assure the correct communication between pollen and pistil. Here we queried the whole transcriptome of Arabidopsis thaliana in order to identify genes with specific reproductive functions. We used the ATH1 whole genome array to profile wild-type unpollinated pistils and unfertilized ovules in comparison with the expression profile of pistils 0.5, 3.5 and 8.0 hours after pollination KEYWORDS: time course Flowers at the developmental stage 12c were emasculated 24 hours before pollination. Pistils were collected at 0, 0.5, 3.5 and 8 Hours After Pollination (HAP) and immediately frozen in liquid nitrogen. Unfertilized ovules were collected by the funiculus from dissected UP and immediately frozen in liquid nitrogen. To minimize biological variation 20 pistils were collected from a minimum of 10 plants and for ovule isolation 50 pistils were used from about 30 plants to isolate approximately 1500 ovules for each replicate experiment.
Project description:Background: The carpel margin meristem is a vital multi-potent structure located in the medial domain of the Arabidopsis thaliana gynoecium, the female floral reproductive organ. The carpel margin meristem generates ovules that upon fertilization become seeds. The molecular mechanisms that specify this meristematic region and regulate its organogenic potential are poorly understood. Here, we present an analysis of the transcriptional profile of the medial domain of the Arabidopsis gynoecium highlighting the developmental stages that immediately proceed ovule initiation, the earliest stages of seed development. Results: Using a floral synchronization system and a SHATTERPROOF2 domain-specific reporter construct, paired with fluorescence-activated cell sorting, we assayed the transcriptome of the gynoecial medial domain with temporal and spatial precision. Our analysis reveals a set of genes that are differentially-expressed within the SHATTERPROOF2 expression domain that marks portions of the developing medial domain. Many members of this gene set have been shown previously to function during the development of medial domain-derived structures, including the ovules, thus validating our approach. Other uncharacterized members of this gene set, including a set of differentially-expressed cis-natural antisense transcripts, are potential novel regulators of medial domain development and candidates for future functional studies. Several members of the REM family of transcriptional regulators were enriched in the SHP2-expressing cell population including a previously unrecognized REM family member (At5g60142). Analysis of the abundance of specific transcriptional isoforms identified genes that may exhibit “isoform switching” behavior during gynoecial development. Conclusions: This data set provides genome-wide transcriptional insight into the development of the gynoecial medial domain that contains the carpel margin meristem, an important reproductive structure that gives rise to the ovules in Arabidopsis thaliana.
Project description:Part of a set of highly integrated epigenome maps for Arabidopsis thaliana. Keywords: Illumina high-throughput transcriptome sequencing
Project description:Background: The carpel margin meristem is a vital multi-potent structure located in the medial domain of the Arabidopsis thaliana gynoecium, the female floral reproductive organ. The carpel margin meristem generates ovules that upon fertilization become seeds. The molecular mechanisms that specify this meristematic region and regulate its organogenic potential are poorly understood. Here, we present an analysis of the transcriptional profile of the medial domain of the Arabidopsis gynoecium highlighting the developmental stages that immediately proceed ovule initiation, the earliest stages of seed development. Results: Using a floral synchronization system and a SHATTERPROOF2 domain-specific reporter construct, paired with fluorescence-activated cell sorting, we assayed the transcriptome of the gynoecial medial domain with temporal and spatial precision. Our analysis reveals a set of genes that are differentially-expressed within the SHATTERPROOF2 expression domain that marks portions of the developing medial domain. Many members of this gene set have been shown previously to function during the development of medial domain-derived structures, including the ovules, thus validating our approach. Other uncharacterized members of this gene set, including a set of differentially-expressed cis-natural antisense transcripts, are potential novel regulators of medial domain development and candidates for future functional studies. Several members of the REM family of transcriptional regulators were enriched in the SHP2-expressing cell population including a previously unrecognized REM family member (At5g60142). Analysis of the abundance of specific transcriptional isoforms identified genes that may exhibit âisoform switchingâ behavior during gynoecial development. Conclusions: This data set provides genome-wide transcriptional insight into the development of the gynoecial medial domain that contains the carpel margin meristem, an important reproductive structure that gives rise to the ovules in Arabidopsis thaliana. Four samples (YFP-positive, YFP-negative, all-sorted, and non-sorted). Four biological replicates of each sample (except YFP-positive, collected in triplicated). Strand-specific libraries were sequenced (single-end) in two lanes of a HiSeq 2500 Illumina flow cell (each lane was later analyzed as a technical replicate).
Project description:Part of a set of highly integrated epigenome maps for Arabidopsis thaliana. Keywords: Illumina high-throughput transcriptome sequencing Transcriptome sequencing of wildtype Arabidopsis plants (Columbia-0), and met1, drm1 drm2 cmt3, and ros1 dml2 dml3 null mutants using the Illumina Genetic Analyzer.
Project description:Purpose: The goal of this study is to compare NGS-derived Arabidopsis thaliana transcriptome profiling (RNA-Seq) obtained from tissues infected with a variety of Pseudomonas syryingae effector mutants to better understand their impact on their host's transcriptional program. We report the application of RNA-sequencing technology (Illumina Hi-Seq) for high-throughput profiling Arabidopsis thaliana transcriptomes upon infection with the model pathogen Pseudomonas syringae (wild-type and mutants).
Project description:High-throughput sequencing of Arabidopsis thaliana endogenous small RNAs by 454 pyrosequencing. Keywords: high-throughput sequencing