Lotus japonicus response to Nod factor: wild type and nfre mutants
ABSTRACT: For RNA sequencing, 3 days seedlings were moved to agar plates supplemented with 1/4 B&D media and susceptible zone of 14 days-old plants was harvested after specific treatment with water (Mock treated) or M.loti Nod factor 10-8M (NF). The total RNA was isolated from the susceptible zone (15 mm root pieces) using Nucleo spin RNA plant (Macherey-Nagel). Total RNA (> 0.8 g) from two biological replicas per sample was used by GATC Biotech (Germany) to prepare random primed cDNA library and for sequencing with Illumina HiSeq: read length 1 x 50bp. Gifu- L. japonicus wild-type, 4820- nfre-1 allele of Nfre, 38534- nfre-2 allele of Nfre.
Project description:Lotus japonicus is a model legume broadly used to study transcriptome regulation under different stress conditions and microorganism interaction. Understanding how this model plant protects itself against pathogens will certainly help to develop more tolerant cultivars in economically important Lotus species as well as in other legumes. In order to uncover the most important defense mechanisms activated upon bacterial attack, we explored by microarray analysis the transcriptome regulation occurring in the phenotypically contrasting ecotypes MG-20 and Gifu B-129 of L. japonicus after inoculation with the non-pathogenic strain Pseudomonas syringae DC3000 pv. tomato.
Project description:Gene expression was studied at the periphery, an intermediate zone, and the centre of wild-type and ∆flbA colonies using Affymetrix A. niger whole genome microarrays. We used Affymetrix GeneChip A. niger Geome Arrays and identifed up- and down-regulated genes that may account for the differences between wild-type and ΔflbA colonies. RNA was isolated from a biological duplicate of concentric zones of 7-day old wildtype and mutant colonies. Zone 1 represented the most central zone, zone 3 an intermediate zone, and zone 5 the peripheral zone. Mycelium of the distinct zones was harvested from three colonies.
Project description:We use the gowth zone of the maize leaf as a model system to study the growth reduction in response to drought stress. The spatial gradient and the relatively large size of the maize leaf allowed us to sample at a subzonal rezolution and to examine different developmental stages at the same time. We compared the response to different levels of drought stress (mild and severe) of proliferating (meristem), expanding (elongation zone) and differentiated (mature zone) tissue. Three separate loop designs were used for the three zones of the maize leaf (meristem, elongation zone, and mature zone). In each loop three treatments were contrasted (control, mild stress, and severe stress). Four biological replicates were used for each zone/condition (4 replicates x 3 zones x 3 conditions = 36 samples).
Project description:Zone 1 and zone 3 areas were captured from mouse liver sections by laser capture microdissection Total RNA was isolated and gene expression profiles were determined using microarrays Six C3H/HeNCrlBR (C3H) mice, 3 male and 3 female mice were used. They were not treated in any way.
Project description:Background: While stressful events are recognized as an important cause of major depressive disorder (MDD), some individuals exposed to life stressors maintain normal psychological functioning. Although the molecular mechanism(s) underlying this phenomenon remain unclear, abnormal transmission and plasticity of hippocampal synapses may play a key role in the pathoetiology of MDD. Methods: A chronic mild stress (CMS) protocol was applied to separate susceptible and unsusceptible rat subpopulations. Proteomic analysis using an isobaric tag for relative and absolute quantitation (iTRAQ) coupled with tandem mass spectrometry was performed to identify differential proteins in enriched hippocampal synaptic junction preparations. Results: A total of 4318 proteins were quantified, and 89 membrane proteins were present in differential amounts. Of these, SynaptomeDB identified 81 (91%) having a synapse-specific localization. The unbiased profiles identified several candidate proteins within the synaptic junction that may be associated with stress vulnerability or insusceptibility. Subsequent functional categorization revealed that protein systems particularly involved in membrane trafficking at the synaptic active zone exhibited a positive strain as potential molecular adaptations in the unsusceptible rats. Moreover, through STRING and immumoblotting analysis, membrane-associated GTP-bound Rab3a and Munc18-1 appear to co-regulate syntaxin-1/SNAP25/VAMP2 assembly at the hippocampal presynaptic active zone of unsusceptible rats, facilitating SNARE-mediated membrane fusion and neurotransmitter release, and may be part of a stress-protection mechanism in actively maintaining an emotional homeostasis. Conclusions: The present results support the concept that there are a range of potential protein adaptations in the hippocampal synaptic active zone of unsusceptible rats, revealing new investigative targets that may contribute to a better understanding of stress insusceptibility.
Project description:Fusarium oxysporum f. sp. conlutinans (Foc) is a serious root-invading and xylem-colonizing fungus that causes yellowing in Brassica oleracea. To comprehensively understand the interaction between F. oxysporum and B. oleracea, composition of the xylem sap proteome of the non-infected and Foc-infected plants was investigated in both resistant and susceptible cultivars using liquid chromatography-tandem mass spectrometry (LC-MS/MS) after in-solution digestion of xylem sap proteins. Whole genome sequencing of Foc was carried out and generated a predicted Foc protein database. The predicted Foc protein database was then combined with the public B. oleracea and B. rapa protein databases downloaded from Uniprot and used for protein identification. About 200 plant proteins were identified in the xylem sap of susceptible and resistant plants. Comparison between the non-infected and Foc-infected samples revealed that Foc infection causes changes to the protein composition in B. oleracea xylem sap where repressed proteins accounted for a greater proportion than those of induced in both the susceptible and resistant reactions. The analysis on the proteins with concentration change >=2 fold indicated a large portion of up- and down-regulated proteins were those acting on carbohydrates. Proteins with leucine-rich repeats and legume lectin domains were mainly induced in both resistant and susceptible system, so was the case of thaumatins. Twenty-five Foc proteins were identified in the infected xylem sap and ten of them were cysteine-containing secreted small proteins that are good candidates for virulence and/or avirulence effectors. The findings of differential response of protein contents in the xylem sap between the non-infected and Foc-infected samples as well as the Foc candidate effectors secreted in xylem provide valuable insights into B. oleracea-Foc interactions.
Project description:Wood is formed by the differentiation of cells from the vascular cambium and it is an important source for pulp, paper and bioenergy production. However there is little information about the vascular cambium at the molecular level, particularly in response to seasonality in tropical regions. We used three different molecular approaches: transcripts, proteome and metabolome to characterize the seasonal alterations in the primary metabolism of the eucalyptus cambial zone. Based on 2-DE analysis, 71 proteins were differentially expressed.