Project description:Physcomitrella patens gametophores were treated with exudates from the arbuscular mycorrhiza fungi (AMF) Rhizophagus irregularis (formerly known as Glomus intraradices) and Gigaspora margerita for one hour and 24 hours.
Project description:Protonema of Physcomitrella patens Pp_A_Doko#069 (DOKO69; Koprivova et al., 2004) grown in liquid cultures with Knop medium (Reski and Abel, 1985) were harvested 6 days after last subculture and setting density to 120 mg/L dry weight.
Project description:***This experiment was updated on 26-02-2016 to correct the accidental mis-labelling of some raw data files. For data downloaded before this date, please note that the file labelled as Caulonema_a.pair contains the raw data for Chloronema A and the file labelled as Chloronema_a.pair contains the data for Caulonema A. ***** We analyzed the transcriptional profile of all phases in the life cycle of P. patens, including all major tissues and 4 different sporophyte developmental stages. For this analysis we took advantage of a custom designed NimbleGene microarray based on genome version 1.6, and developed adequate protocols for the isolation and purification of mRNA directly from mechanically purified cells of chloronema, caulonema, rhizoids, gametophores, sporophytes, spores, archegonia and protoplasts.
Project description:We experimentally generated a genome-wide gene expression data. To this end, we first collected samples of three important haploid developmental stages, specifically germinating spores (gametophyte_1), protonemata (gametophyte_2) and young gametophores (gametophyte_3) (four biological replicates each). We also collected three developmental stages of the diploid phase (sporophyte), specifically sporophytes shorter than 5mm (sporophyte_1), elongated needle-like sporophytes (sporophyte_2), and sporophytes with swollen capsules (sporophyte_3)
Project description:7 days old protonema of Physcomitrella patens Wildtype and two transgenic lines, ΔPpcmt line 281 (Noy-Malka et al. 2014; IMSC accession 40738) and ΔPpmet line 5 (Yaari et al. 2015; IMSC accession 40758) grown on BCDAT solid medium.
Project description:We studied peptide generation process in moss Physcomitrella patens. For this purpose, analysis of peptidome and transcriptome of two different life forms (gametophores and protonema) and also of protoplasts were made.
Project description:Brown algae (Phaeophyceae) are complex photosynthetic organisms with a very different evolutionary history to green plants, to which they are only distantly related. These seaweeds are the dominant species in rocky coastal ecosystems and they exhibit many interesting adaptations to these, often harsh, environments. The brown algae are also important because they are one of only a very small number of eukaryotic lineages that have evolved complex multicellularity. This work used whole genome tiling array approach to generate a comprehensive transcriptome map of the filamentous seaweed Ectocarpus siliculosus (Dillwyn) Lyngbye, a model organism for the brown algae. Keywords: high-resolution tiling array, brown algae, ectocarpus The slides were hybridised with two, labelled samples: 1) a mixture of labelled cDNA corresponding to RNA samples from mature sporophytes and gametophytes and from immature sporophytes stressed either in high salt medium or by addition of hydrogen peroxide and 2) genomic DNA as a control.
Project description:We next sought to identify the transcriptional program that differentiates IL-9+Th2 cells from “conventional” Th2 cells. To this end, we selected representative Th1, Th17, Th2, and IL-9+Th2 clones (Figure 5A) and determined their transcriptome in the resting state and at different time points after activation using RNAseq. Peripheral Blood Mononuclear Cells (PBMC) were isolated by Ficoll-Plaque Plus (GE Healthcare, UK) density gradient centrifugation. Human CD4+ T cells were isolated from PBMC by EasySep positive selection kit (Stemcell Technologies) according to manufacturer’s instruction. Positively selected CD4+ T cells were washed with PBS and stained for subsequent Th cell subset sorting. Memory Th cell subsets were sorted to over 90% purity according to their expression of chemokine receptors from CD45RA-CD25-CD8-CD3+ cells: Th1(CXCR3+CCR8-CCR6-CCR4-), Th2 (CXCR3-CCR8-CCR6-CCR4+), Th17 (CXCR3-CCR8-CCR6+CCR4+), Th9 (CXCR3-CCR8+CCR6-CCR4+). Single cell Th subset clones were directly sorted into 96well plate according to their expression of chemokine receptors (see above). Single cell clones were expanded and maintained by periodic restimulation with PHA (phytohaemaglutinine, 1 µg/ml, Sigma-Chemicals) and irradiated allogenic feeder cells (5x104/well) in culture medium. T cells were polyclonally activated using beads coated with antibodies against CD3, CD2, and CD28 (T cell/bead = 2:1, human T cell activation/expansion Kit, Miltenyi). Cell cultures were sampled before activation (time 0h) and 2, 4, 6, 9, 12, 24, and 48 hours after activation.