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:Physcomitrella patens reproductive organs (archegonia and antheridia) were mechanically isolated for transcriptome profiling. Comparison between wt and Ppglr1/2, a glutamate receptor double knock-out mutant, was done to identify deferentially expressed genes. A custom designed NimbleGene microarray based on genome version 1.6 was used for this study.
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:The data is a result of a large laboratory experiment targeting interactive effects of salinity and temperature on the transcriptomic level in algae from two populations of an ecologically relevant kelp species (Laminariales), Saccharina latissima. Research interest in S. latissima has recently been increasing given its importance as ecosystem engineer along temperate rocky shores in the Atlantic Ocean, and its growing potential in industrial applications such as aquaculture, pharmaceutics, food and feed. Young sporophytes of S. latissima were raised from stock cultures of clonal male and female gametophytes at the Alfred-Wegener-Institute Helmholtz Centre for Polar and Marine Research. Cultures originated from sporophyte collected at Kongsfjorden (79°N, 11°E; Spitsbergen, Norway) and at Roscoff (48° 43′ 39″ N, 3° 59′ 13.2″ W; Brittany, France). Sporophytes of both populations were grown aerated in glass beakers at 8°C and under a photon fluence rate of 20 µmol photons m–2 s–1 of photosynthetically active radiation with a 18 h light: 6 h dark photoperiod and were cultivated in sterile seawater enriched with Provasoli (Starr & Zeikus 1993) with an absolute salinity (SA) of ~30 during three months. At the start of the experiment, sporophytes were either kept at 8°C (or transferred to 0°C and 15°C) in temperature controlled rooms. After one week, per each temperature, sporophytes were divided into a low salinity treatment of SA 20 or kept at the control salinity (SA 30).
Project description:To identify transcription factor (TF) genes exhibiting preferential expression during SE, expression profiles of 1,880 TFs were compared in two genotypes with largely different capacities for SE induction in IZE culture on auxin medium, namely the highly embryonic Col-0 accession and the tanmei mutant unable to form somatic embryos. Our study revealed 729 TFs whose expression changes during the 10-day incubation period of SE; 141 TFs displayed distinct differences in expression patterns in embryogenic versus non-embryogenic cultures. This study provides comprehensive data focused on the expression of TF genes during SE and provides guidelines for further research on functional genomics of SE. The experiment was designed to monitor the expression of 1,880 TF genes at three distinctive stages of IZE-derived embryogenic culture, which refer to: (i) freshly isolated explants competent to undergo embryogenic transition (0 d), (ii) explant tissue subjected to SE induction (5 d), and (iii) the advanced phase of embryogenesis related to somatic embryo formation (10 d).
Project description:Bryophytes are the most basal of the extant land plants. A major feature of these plants is the biphasic alteration of generations between a dominant haploid gametophyte and a minor diploid sporophyte phase. To analyse the differences in the transcriptome of the early gametophyte (protonema) and early and mid-sporophyte phases of the moss Physcomitrella patens, microarray gene expression profiles were performed using dissected sporophyte tissue. Through further analysis the early and mid-sporophyte phases were compared. RNA isolated from the Gametophytic protonemal tissue was hybridised to six microarrays. Each microarray was hybridised with RNA from a separate biological replicate. Three of these microarrays were co-hybridised with RNA isolated from early sporophytes. With the third gametophyte biological replicate and early sporophyte replicate a dye swap was carried out. The remaining three microarrays hybridised with RNA from the gametophytes were co-hybridised with RNA from mid-sporophytic tissue. A dye swap was carried out on the sixth gametophyte replicate and third mid-sporophyte replicate.To meet the quality requirements for the microarray experiment, at least 400 sporophytes were used per extraction. Three or four RNA extracts were then pooled for further precipitation to maximise purity and concentration. Up to 1600 sporophytes were harvested to prepare sufficient RNA for each microarray replicate. In bioinformatic analysis the channels were split into individual channels and the early and mid-sporophyte were compared.
Project description:The aim of this experiment was to compare transcript abundances in parthenotes (i.e. organisms derived by parthenogenetic development of gametes) of two life cycle mutants of the brown alga Ectocarpus siliculosus with transcript abundances in the wild type sporophyte and gametophyte generations. This is of interest because the two mutations, immediate upright (imm) and ouroboros (oro), cause partial and almost complete hometic conversion, respectively, of the sporophyte into the gametophyte. imm parthenotes exhibit gametophyte-like morphology during early development but remain sporophytes in functional terms (they do not produce gametes) whereas oro parthenotes behave as functional gametophytes and are morphologically indistinguishable from gametophytes apart from the appearance some minor sporophyte-like features very early in development in some individuals. To minimise genetic background effects the samples for this experiment were derived from a segregating population derived from an imm/IMM oro/ORO sporophyte. Four classes of gametophyte were derived from this sporophyte were IMM/ORO (wild type), imm/ORO, IMM/oro and imm/oro. Parthenomes were bulked to provide a wild type sporophyte sample, samples corresponding to the two individual mutants, plus the double mutant. A wild type gametophyte sample was also compared for comparison. Hybridisations with cDNA derived from these five samples were carried out using a NimbleGen expressed-sequence-tag-(EST-)based microarray carrying probes corresponding to 10,600 of the 16,256 genes identified in the Ectocarpus genome.
Project description:Effect of PAR and temperature stress on the gene expression Saccharina latissima. Total RNA of stress treatments (low PAR 2° and 17°C, high PAR 2° and 17°C) was hybridized against the control treatment (low PAR 12°C); hybridizations were carried out in 4 replicates.
Project description:Heterotrimeric G proteins mediate crucial and diverse signaling pathways in eukaryotes. To gain insights into the regulatory modes of the G protein and the co-regulatory modes of the G protein and the stress hormone abscisic acid (ABA), we generated and analyzed gene expression in G protein subunit single and double mutants of the model plant Arabidopsis thaliana. Through a Boolean modeling approach, our analysis reveals novel modes of heterotrimeric G protein action. Keywords: transcriptome analysis; G protein subunit mutants; abscisic acid (ABA) Microarray data were generated from four genotypes (wild type, gpa1-4 mutant, agb1-2 mutant, agb1-2 gpa1-4 double mutant) with or without ABA treatment. Arabidopsis plants were grown in growth chambers with an 8 hr light/16hr dark. Three hundred Arabidopsis leaves excised from 60-70 five-week-old plants were used as the starting material for each guard cell microarray. Ten mature leaves taken from 3-4 plants grown side-by side with the plants for guard cell isolation were used for each leaf sample. Excised leaf and isolated guard cell samples were treated with ABA (50 μM) or EtOH (solvent control) for 3 hrs. For each type of sample (guard cells or leaves), three independent biological replicates were performed, resulting in a total of 48 microarray hybridizations (2 sample types ´ 4 genotypes ´ two treatments ´3 replicates).