Project description:Rapid, reversible induction of crassulacean acid metabolism in conjunction with pronounced reconfiguration of carbon metabolism enables T. triangulare to survive cycles of severe drought. RNA was extracted from leaves of Talinum triangulare plants (3 biological replicates) after plants were grown under 0, 4, 9 and 12 days of drought as well as after 2 days of re-watering. Paired-end samples were used for purposes of creating a de novo transcriptome assembly.
Project description:What proteins exist in the epidermis layer and how they compare to the proteins in the surrounding mesophyll cells represents a major knowledge gap for how CAM plants thrive in environments with high temperatures and long periods of severe water deficit. Therefore, we performed large-scale proteomics to characterize proteins in epidermis and mesophyll cells from leaves of the constitutive CAM plant Kalanchoë fedtschenkoi.
Project description:Bienertia sinuspersici performs C4 photosynthesis without Kranz anatomy through subcellular compartmentalization of carbon fixation within individual cells. In this species, central compartment chloroplasts (C) and peripheral chloroplasts (P) collaborate with cytosolic and mitochondrial components in a NAD-ME type C4 cycle. How the two functionally different chloroplast types can develop within individual cells and the mechanism of import of nuclear encoded, plastid targeted proteins are currently unknown. We used 454 sequencing in combination with large scale label-free proteomics to determine the distribution of photosynthesis-related proteins. Subcellular localization of 169 protein was determined through comparison of protein abundance in four different subcellular fractions. 39 out of the 120 chloroplastic proteins showed differential accumulation between the two chloroplast types. Rubisco, RPP regenerative phase and PSII related proteins accumulated in C chloroplasts whereas C4 related proteins and the NDH complex were more abundant in P chloroplasts. Comparison of transit peptides of differential accumulating proteins indicated no obvious sequence homology or similarities in physico-chemical properties between members of the same group. Protein composition analysis of the central compartment indicated that mitochondria and peroxisomes are the only major components besides chloroplasts in this compartment. The combined information from subcellular and developmental protein profiling was used to generate a first draft of the protein machinery involved in single-cell C4 photosynthesis.
Project description:Plants of two non-restorer varieties of hexaploid winter wheat (Astoria, Grana) and two restorers ones (Patres and Primépi) were used to Representational Difference Analysis (cDNA-RDA) to identify effective Rf (fertility restorer) genes. Anthers at the microspore stage were used as testers while anthers excised at the stage of pollen tetrads were selected as driver. The tester:driver ratio in three subsequent rounds of subtractive hybridization increased from 1:50, and 1:400 to 1:200000. Differential products obtained in the third subtraction of the size range from 200 to 800 bp were sequenced.
Project description:Comparison of BOLITA mutant leaves (gain-of-function mutant) vs. wild type leaves, grown in the same conditions. The BOLITA (BOL) gene (At1g24590), an AP2/ERF transcription factor, was characterized with the help of an activation tag mutant and overexpression lines in Arabidopsis and tobacco. The leaf size of plants overexpressing BOL was smaller than wild type plants due to a reduction in both cell size and cell number. Moreover, severe overexpressors showed ectopic callus formation in roots. Accordingly, global gene expression analysis using the overexpression mutant reflected the alterations in cell proliferation, differentiation and growth through expression changes in RBR, CYCD, and TCP genes, as well as genes involved in cell expansion (i.e. expansins and the actin remodeling factor ADF5). Furthermore, the expression of hormone signaling (i.e. auxin and cytokinin), biosynthesis (i.e. ethylene and jasmonic acid) and regulatory genes was found to be perturbed in bol-D mutant leaves. The gene is expressed at the shoot apical meristem, and more intensely at leaf primordia. It is also expressed at very young leaves, and flower buds. The gene is closely related to DORNROSCHEN.
Project description:The growth and shape of plants is mainly defined by the properties of their cell walls, which dynamically adapt to internal and external cues. Therefore, the cell wall is under constant surveillance to relay feedback information to the cell interior. However, very little is known about how cell wall signaling is integrated with intracellular growth regulation. Here, we have identified a receptor-like protein, RLP44, which conveys information from the cell wall to the brassinosteroid hormone signaling pathway by interacting with the regulatory receptor-like kinase BAK1. This conditional RLP44-mediated signaling activation is required for normal development and stress responses, requires functional brassinosteroid receptor, but is partially independent of the hormone ligand. We sought to identify transcriptional changes in the PMEIox transformant in which an altered pectin modification leads to an induction of the brassinosteroid signalling pathway. In addition we determined which fraction of these expression changes was reverted in the cnu1 and cnu2 suppressor mutants of PMEIox, respectively. Complete transcriptome analysis was performed on Col-0 wild-type as well as the PMEIox transformant and the cnu1 and cnu2 suppressor mutant seedlings grown in the dark for four days. 3 Biological replicates of etiolated seedlings grown for 4 days in the dark were harvested on successive weeks.
Project description:When macrophages encounter pathogens, they transiently induce an orchestrated cascade of pro- and anti-inflammatory genes. To obtain a precise picture of transcriptome-wide mRNA expression patterns, we performed RNA-Seq of total RNA at a high temporal resolution during the first two hours of macrophage activation. We systematically analyzed the contribution of translational regulation to the early phase of macrophage activation. While the expression of most cytokines is pre-dominanatly regulated by changes in mRNA levels, de-repression of translation was found to permit expression of many feedback inhibitors of the inflammatory response. Expression profiles of LPS-treated Raw264.7 cells (0, 15, 30, 45, 60, 75, 90 and 120 min after stimulation) were generated by deep sequencing using Illumina HiSeq 2000.
Project description:Plants of two non-restorer varieties of hexaploid winter wheat (Astoria, Grana) and two restorers ones (Patres and Primépi) were used to identify effective Rf (fertility restorer) genes by next generation sequencing on whole transcriptomes (RNA-seq).
Project description:RNA from wheat lodicules in two highly-chasmogamous (HCH) (Piko and Poezja) and two low-chasmogamy (LCH) (Euforia and KWS Dacanto) varieties at two developmental stages - pre-flowering and early flowering were used to Representational Difference Analysis (cDNA-RDA). Lodicules at the pre-flowering were used as testers while the lodicules early flowering were selected as driver probes. This procedure results in accumulation of unique sequences upregulated in pre-flowering. The analysis was taken in three replicates for each variety (Piko, Poezja, Euforia, KWS Dacanto). The tester:driver ratio in three subsequent rounds of subtractive hybridization increased from 1:50, and 1:400 to 1:200000. Difference products obtained from the third subtraction in the size range from 200 to 800 bp were sequenced.
Project description:Dynamic acclimation of photosynthesis plays an important role in increasing plant fitness under variable light environments. Acclimation is mediated by a glucose-6-phosphate/phosphate translocator, GPT2. This study examined whether plants lacking GPT2, which are defective in acclimation to increases in light, are more susceptible to oxidative stress. To understand this, we used the model plant of Arabidopsis thaliana (accession Wassilewskija-4 (Ws-4)) and compared this to mutants lacking GPT2. Plants were grown at low light (100 μmol m−2 s−1) for 7 weeks. For acclimation experiments, a set of plant from low light was transferred to 400 μmol m−2 s−1 for 7 days. Microarray analysis showed that gpt2 plants showed a greater induction of stress related genes relative to WT.