43 research outputs found

    Subcellular localization and functional characterization of the Arabidopsis thaliana homologue of cellular repressor of E1A-stimulated genes (CREG)

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    „Cellular repressor of E1A-stimulated genes“ (CREG) ist ein Phosphoglykoprotein in den Lysosomen tierischer Zellen. Ziel dieser Arbeit war es , Einsicht in die subzelluläre Lokalisation und Funktion von CREG in pflanzlichen Zellen zu erlangen. Hierfür wurden zwei Konstrukte kreiert, die durch konfokale Laser-Scanning Mikroskopie in den Zellen von Nicotiana benthamiana und Arabidopsis thaliana lokalisiert wurden. Die Analyse ergab, dass sich aCREG aller Wahrscheinlichkeit nach in den Vakuolen pflanzlicher Zellen befindet. Für die funktionelle Analyse von aCREG wurden A. thaliana Wildtyp Pflanzen mit zwei aCREG knock-down Linien unter abiotischen Stress-Bedingungen verglichen. Die Ergebnisse deuten darauf hin, dass aCREG keinen direkten Einfluss auf die Reaktion von Pflanzen auf abiotischen Stress ausübt.“Cellular repressor of E1A-stimulated genes” (CREG) is a phosphoglycoprotein found in the lysosomes of mammalian cells. The aim of this work was to investigate the subcellular localization of CREG in plant cells and to get an insight into its functions there. For subcellular localization studies in Nicotiana benthamiana and Arabidopsis thaliana, two constructs, that were analysed by confocal laser scanning microscopy, were created. The analysis shows, that aCREG is localized in the vacuoles of plant cells. For functional analysis of aCREG in A. thaliana, wildtype seedlings were compared to two aCREG knock-down lines under abiotic stress conditions. The results indicate that aCREG is dispensible for the abiotic stress response of plants

    Revised sequence and annotation of the Rhodobacter sphaeroides 2.4.1 Genome

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    The DNA sequences of chromosomes I and II of Rhodobacter sphaeroides strain 2.4.1 have been revised, and the annotation of the entire genomic sequence, including both chromosomes and the five plasmids, has been updated. Errors in the originally published sequence have been corrected, and ∼11% of the coding regions in the original sequence have been affected by the revised annotation

    Microbiomes of Velloziaceae from phosphorus-impoverished soils of the campos rupestres, a biodiversity hotspot

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    The rocky, seasonally-dry and nutrient-impoverished soils of the Brazilian campos rupestres impose severe growth-limiting conditions on plants. Species of a dominant plant family, Velloziaceae, are highly specialized to low-nutrient conditions and seasonal water availability of this environment, where phosphorus (P) is the key limiting nutrient. Despite plant-microbe associations playing critical roles in stressful ecosystems, the contribution of these interactions in the campos rupestres remains poorly studied. Here we present the first microbiome data of Velloziaceae spp. thriving in contrasting substrates of campos rupestres. We assessed the microbiomes of Vellozia epidendroides, which occupies shallow patches of soil, and Barbacenia macrantha, growing on exposed rocks. The prokaryotic and fungal profiles were assessed by rRNA barcode sequencing of epiphytic and endophytic compartments of roots, stems, leaves and surrounding soil/rocks. We also generated root and substrate (rock/soil)-associated metagenomes of each plant species. We foresee that these data will contribute to decipher how the microbiome contributes to plant functioning in the campos rupestres, and to unravel new strategies for improved crop productivity in stressful environments6COORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR - CAPESFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP88881.068071/2014-012016/23218-0Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP) [2016/23218-0]; U.S. Department of Energy Joint Genome Institute (DOE-JGI)United States Department of Energy (DOE) [CSP 503222]; Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES)CAPES [88881.068071/2014-01]; FAPESPFundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP) [2018/04240-0]; CAPESCAPES; Office of Science of the U.S. Department of EnergyUnited States Department of Energy (DOE) [DE-AC02-05CH11231

    Systems analysis of lactose metabolism in Trichoderma reesei identifies a lactose permease that is essential for cellulase induction.

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    Trichoderma reesei colonizes predecayed wood in nature and metabolizes cellulose and hemicellulose from the plant biomass. The respective enzymes are industrially produced for application in the biofuel and biorefinery industry. However, these enzymes are also induced in the presence of lactose (1,4-0-ß-d-galactopyranosyl-d-glucose), a waste product from cheese manufacture or whey processing industries. In fact, lactose is the only soluble carbon source that induces these enzymes in T. reesei on an industrial level but the reason for this unique phenomenon is not understood. To answer this question, we used systems analysis of the T. reesei transcriptome during utilization of lactose. We found that the respective CAZome encoded all glycosyl hydrolases necessary for cellulose degradation and particularly for the attack of monocotyledon xyloglucan, from which ß-galactosides could be released that may act as the inducers of T. reesei's cellulases and hemicellulases. In addition, lactose also induces a high number of putative transporters of the major facilitator superfamily. Deletion of fourteen of them identified one gene that is essential for lactose utilization and lactose uptake, and for cellulase induction by lactose (but not sophorose) in pregrown mycelia of T. reesei. These data shed new light on the mechanism by which T. reesei metabolizes lactose and offers strategies for its improvement. They also illuminate the key role of ß-D-galactosides in habitat specificity of this fungus

    Figure 2

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    <p>(a) Growth comparison of QM 9414 to the Δ3405 strain on minimal media supplemented with D-glucose, lactose and D-galactose. Pictures were taken on day 4. (b-g) Comparison of strains QM 9414 (grey), Δ3405 (white) and <i>ptef::3405</i> (black) pregrown in minimal medium supplemented with 1% glycerol and,after washing of the mycelium extensivly with sterile water, transferred to minimal medium containing 1% lactose. (b) Accumulation of <i>cel6a</i>, <i>cel7a</i> and <i>xyr1</i> transcripts measured by qPCR. Samples were taken 5, 8, 16 and 24 h after replacement to 1% lactose. The expression is given in relation to <i>tef1</i> gene expression, where the <i>tef1</i> expression value equals one. Mean values ± SD of three independent experiments are shown. (c) Lactose concentration in the medium during growth of QM 9414 (filled diamonds), Δ3405 (empty circles) and <i>ptef::3405</i> (empty squares) on 1% lactose. (d) Biomass formation of strains QM 9414 (filled diamonds), Δ3405 (empty circles) and <i>ptef::3405</i> (empty squares) on 1% lactose. Mean values ± SD of three independent experiments are given. (e) SDS-PAGE analysis of culture filtrates from QM 9414, Δ3405 and <i>ptef::3405.</i> Samples taken at 16 and 24 h after replacement to lactose. (f) ß-galactosidase activity determined with <i>o</i>-nitrophenyl-ß-D-galactopyranoside as the substrate. Error bars indicate the standard deviation of three independent experiments.</p
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