ABSTRACT: The worldwide pest grape phylloxera (Daktulosphaira vitifoliae Fitch) is threatening vititulure. We investigated the compatible interaction with the host Teleki 5C (Vitis berlandiere Planch. X V. riparia Michx.) by investigating differential gene expression analysis using a custom made microarray. Samples (root tips and nodosities) were obtained from not infected and infected one-node cuttings four weeks after inoculation with 60 sibling phylloxera eggs. Four independent biological replicates each were analysed using a doul-color microarray experiment. Dye-swap hybridizations were performed. In total eight microarray were used for hybridization.
Project description:Transcriptional expression in the rachis of pre-sympotmatic (T1) and symptomatic (T2) berry shrivel (BS) clusters in comparison to control samples. Samples were collected from a vineyard in Mailberg, Lower Austria, in 2011 from cultivar Blauer Zweigelt (Vitis vinifera) grafted on Kober 5BB. The objetive of the study was to identify metabolic processes changed in the rachis due to BS induction and symptom development
Project description:Cellular identity is ultimately dictated by the interaction of transcription factors with regulatory elements (REs) to control gene expression. Advances in genome-wide epigenome profiling techniques have significantly increased our understanding of cell-specific utilization of REs. However, it remains difficult to dissect the majority of factors that interact with these REs due to the lack of appropriate techniques. To address this issue, we developed a novel epigenetic method termed TINC: TALE-mediated Isolation of Nuclear Chromatin. Using TINC we interrogated the protein complex formed at the Nanog promoter in embryonic stem cells (ESCs) and identified many known interactors as well as numerous previously unknown complex members, including RCOR2. Further interrogation of the role of RCOR2 in the ESC network revealed its involvement in the repression of lineage genes and the fine-tuning of pluripotency genes. Consequently, using the Nanog promoter as a paradigm, we demonstrated the power of TINC to provide important insights into the molecular makeup of transcriptional complexes at individual REs as well as into control of cellular identity in general.
Project description:Herbivory plant-parasite interactions depend on the delivery of effector molecules by the invading insect species. Sedentary gall forming insects, such as grape phylloxera (Daktulosphaira vitifoliae FITCH, Phylloxeridae) secrete multiple effectors into host plant tissues that alter host cellular functions to the benefit of the insect. Analyses revealed 420 putative âDvEffectorsâ were detected in salivary glands, dissected from root-feeding vs. starving D. vitifoliae larvae reared on Teleki 5C (V. berlandieri x V. riparia) under controlled growth conditions (25Â±3Â°C, 60% rH) by proteomic mass spectrometry and in-silico secretory prediction. Sixty-two conserved DvEffectors were shared with the aphid species A. pisum, M. persicae and R. padi including candidate effector proteins involved in feeding site establishment, plant defence suppression and nutrient uptake
Project description:The purpose of this study was to identify genes involved in the skin immune response that changed expression in response to scabies mite infestation. Transcriptomic analysis was undertaken using total RNA extracted from skin biopsies. Four biological replicates for non-infested control group and eight (four for crusted scabies disease phenotype and four for ordinary scabies diseases phenotype) biological replicates for the mite-infested group were used for gene expression analysis at baseline week 0, and at weeks 1, 2, 4 and 8 post-infestation using A-GEOD-16571 Agilent Porcine Gene Expression Microarray 4 x 44K.
Project description:Transcriptional profiling of pooled adult kidneys from transgenic zebrafish expressing human KIT-D816V mutant gene versus wild-type zebrafish was performed. The aim of this experiment was to determine the expression and cellular changes caused by expression of KIT-D816V oncogene in the hematopoietic organ in zebrafish. This transgenic zebrafish represents a model of systemic mastocytosis with many features of this disease present in adults Two-condition experiment, Tg(actb2:KIT-D816V) versus wild-type(AB), one replicate of each prepared from 3 adult kidneys of each genotype and a dye-swap hybridization of these RNA samples.
Project description:Transcriptional profiling of 28 hpf zebrafish transgenic expressing human KIT-D816V mutant gene versus wild-type embryos was performed. The aim of this experiment was to determine the expression changes caused by expression of KIT-D816V oncogene. This transgenic zebrafish represents a model of systemic mastocytosis with many features of this disease present in adults, but no very clear phenotypes in embryos. Thus, we attempted to determine if there are certain expression changes, which can be used as molecular features of active KIT expressed in these embryos. Two-condition experiment, Tg(actb2:KIT-D816V) versus wild-type(AB), 4 replicate experiments with a dye-swap each time
Project description:Calcium-activated potassium channels are important membrane proteins that help in maintaining the cellular physiology and cell signalling. Till date no Calcium-activated potassium channels have been identified in Leishmania donovani. It’s gene was cloned into pET vector and expressed in E. coli BL21 (DE3). The recombinant protein was purified using nickel affinity chromatography.
Project description:We have implemented an integrated Systems Biology approach to analyze overall transcriptomic reprogramming and systems level defense responses in the model plant Arabidopsis thaliana during an insect (Brevicoryne brassicae) and a bacterial (Pseudomonas syringae pv. tomato strain DC3000) attack. The main aim of this study was to identify the attacker-specific and general defense response signatures in the model plant Arabidopsis thaliana while attacked by phloem feeding aphids or pathogenic bacteria. Defense responses and networks, unique and specific for aphid or Pseudomonas stresses were identified. Our analysis revealed a probable link between biotic stress and microRNAs in Arabidopsis and thus opened up a new direction to conduct large-scale targeted experiments to explore detailed regulatory links among them. The presented results provide a first comprehensive understanding of Arabidopsis - B. brassicae and Arabidopsis - P. syringae interactions at a systems biology level. Arabidopsis thaliana (ecotype Colombia-0) seeds were grown in 6-cm-diameter pots filled with a sterile soil mix (1.0 part soil and 0.5 part horticultural perlite), 3 plants per pot. Plants were kept in growth chambers Vötsch VB 1514 (Vötsch Industrietechnik GmbH, Germany) under the following conditions: a 8/16 h (light/dark) photoperiod at 22°C/18°C, 40%/70% relative humidity, and 70/0 μmol m-2s-1 light intensity. After 32 days plants had 8 fully developed leaves. Each plant was infested with 32 wingless aphids [Brevicoryne Brassicae] (4 per leaf), which were transferred to leaves with a fine paintbrush. Infested plants and aphid-free controls were kept in plexiglass cylinders. Plants were harvested 72 h after infestation between the 6th and 8th hour of the light photoperiod. Four biological replicates were prepared from aphid infested and control plants, each sampled from 15 individual plants. Whole rosettes were cut at the hypocotyls and aphids were removed by washing with Milli-Q-filtered water. Differences in transcriptional responses were measured by comparing genes expression of aphid infested plants against non-infested control plants.