Project description:Dissection of the genetic pathways and mechanisms by which anther development occurs in grasses is crucial for both a basic understanding of plant development and for traits of agronomic importance like male sterility. In rice, MULTIPLE SPOROCYTES1 (MSP1), a leucine-rich-repeat receptor kinase, play an important role in anther development by limiting the number of sporocytes. OsTDL1a (a TPD1-like gene in rice) encodes a small protein which acts as a cofactor of MSP1 in the same regulatory pathway. In this study, we analyzed small RNA and mRNA changes in different stages of spikelets from wildtype rice, and from msp1 and ostdl1a mutants. Analysis across different stages of rice spikelets of the small RNA data identified miRNAs demonstrating differential abundances. miR2275 was depleted in the two rice mutants; this miRNA is specifically enriched in anthers and functions to trigger the production of 24-nt phased secondary siRNAs (phasiRNAs) from PHAS loci. We observed that the 24-nt phasiRNAs as well as their precursor PHAS mRNAs were also depleted in the two mutants. Based on comparisons of transcript levels across the spikelet stages and mutants, we identified 22 transcription factors as candidates to have roles specific to anther development, potentially acting downstream of the OsTDL1a-MSP1 pathway. An analysis of co-expression identified three Argonaute-encoding genes (OsAGO1d, OsAGO2b, and OsAGO18) that accumulate transcripts coordinately with phasiRNAs, suggesting a functional relationship. By mRNA in situ analysis, we demonstrated a strong correlation between the spatiotemporal pattern of accumulation of these OsAGO transcripts with previously-published phasiRNA accumulation patterns from maize. Raw files for smallRNA-Seq were not provided by submitter.
Project description:The first GSSM of V. vinifera was reconstructed (MODEL2408120001). Tissue-specific models for stem, leaf, and berry of the Cabernet Sauvignon cultivar were generated from the original model, through the integration of RNA-Seq data. These models have been merged into diel multi-tissue models to study the interactions between tissues at light and dark phases.
Project description:Magnaporthe oryzae snodprot1 homologous protein (MSP1) has been shown to act as a pathogen-associated molecular pattern (PAMPs) and trigger PAMP-triggered immunity (PTI) response involving programmed cell death and expression of various defense-related genes in rice. The involvement of several post-translational modifications (PTMs) in the regulation of plant immune response, especially PTI, during pathogen infection is well established, however, the information on the regulatory roles of these PTMs in response to MSP1-induced signaling in rice is currently elusive. Here, we report the phosphoproteome, ubiquitinome, and acetylproteome to investigate the MSP1-induced PTMs alterations in MSP1 overexpressed rice. Our analysis identified a total of 4,666 PTM modified sites in rice leaves including 4,292 phosphosites, 189 ubiquitin sites, and 185 acetylation sites. Among these, PTM status of 437 phosphorylated, 53 ubiquitinated, and 68 acetylated peptides were significantly changed by MSP1. Functional annotation of MSP1 modulated peptides by MapMan analysis revealed that these were majorly associated with cellular immune responses such as signaling, transcription factors, DNA and RNA regulation, and protein metabolism, among others. Taken together, this study uncovers the MSP1-induced PTMs changes in rice proteins and identified several novel components of rice-MSP1 interaction.
Project description:MSP1 is a Magnaporthe oryzae secreted protein that elicits defense responses in rice. However, the molecular mechanism of MSP1 action is largely elusive. Here, we employed a TMT-based quantitative proteomic analysis of cytoplasmic as well as plasma membrane proteins to decipher the MSP1 induced signalling in rice. This approach led to the identification of 6691 proteins of which 3049 were identified in the plasma membrane (PM) while 3642 were identified in the cytoplasmic fraction. A parallel phosphoproteome analysis led to the identification of 1906 phosphopeptides and integration of proteome and phosphoproteome data showed activation of proteins related to the proteolysis, jasmonic acid biosynthesis, redox metabolism and MAP kinase signaling pathways in response to MSP1 treatment. Further, MSP1 induced phosphorylation of some of the key proteins including RBOHB, MEKK1, MPK3/6, CDPK and CaM suggest activation of PAMP-triggered immunity (PTI) in response to MSP1 treatment. In essence, our results further support the functioning of MSP1 as a PAMP and provide an overview of the MSP1 induced signaling in rice leaves.
Project description:Data analysis is a critical part of quantitative proteomics studies in interpreting biological questions. Numerous computational tools including protein quantification, imputation, and differential expression (DE) analysis were generated in the past decade. However, searching optimized tools is still an unsolved issue. Moreover, due to the rapid development of RNA-Seq technology, a vast number of DE analysis methods are created. Applying these newly developed RNA-Seq-oriented tools to proteomics data is still a question that needs to be addressed. In order to benchmark these analysis methods, a proteomics dataset constituted the proteins derived from human, yeast, and drosophila with different ratios were generated. Based on this dataset, DE analysis tools (including array-based and RNA-Seq based), imputation algorithms, and protein quantification methods were compared and benchmarked. This study provided useful information on analyzing quantitative proteomics datasets. All the methods used in this study were integrated into Perseus which are available at https://www.maxquant.org/perseus.
Project description:We performed RNA-seq and Ribo-seq analyses to elucidate the translation in seeds at 85 and 115 DAF. We also completed a data-independent acquisition (DIA)-based proteomic analysis, while also examining relevant lipid metabolites.
Project description:<p>Gene expression is a biological process regulated at different molecular levels, including chromatin accessibility, transcription, and RNA maturation and transport. In addition, these regulatory mechanisms have strong links with cellular metabolism. Here we present a multi-omics dataset that captures different aspects of this multi-layered process in yeast. We obtained RNA-seq, metabolomics, and H4K12Ac ChIP-seq data for wild-type and mip6delta strains during a heat-shock time course. Mip6 is an RNA-binding protein that contributes to RNA export during environmental stress and is informative of the contribution of post-transcriptional regulation to control cellular adaptations to environmental changes. The experiment was performed in quadruplicate, and the different omics measurements were obtained from the same biological samples, which facilitates the integration and analysis of data using covariance-based methods. We validate our dataset by showing that ChIP-seq, RNA-seq and metabolomics signals recapitulate existing knowledge about the response of ribosomal genes and the contribution of trehalose metabolism to heat stress.</p>
Project description:Pathogen secreted pathogen-associated molecular patterns (PAMPs) play a key role in PAMPs triggered immunity (PTI) in plant for the recognition of pathogens. In rice, fewer PAMPs and their pattern recognition receptors (PRRs) have been characterized during rice-Magnaporthe oryzae interaction. Particularly, recent study was identified M. oryzae snodprot1 homolog known as MSP1, however, the molecular mechanism of MSP1 induced PTI is currently elusive. Therefore, we were generated the MSP1 overexpressed transgenic rice with their subcellular localization in the apoplastic (with signal sequence) and cytoplasmic (without signal sequence) regions, respectively, to examine the functional role of MSP1 in rice. Here, we employed a tandem mass tag (TMT)-based quantitative membrane proteomic analysis to decipher the potential interaction PRRs and MSP1-induced downstream signaling. This approach led to the identification of 8,033 proteins and sequential statistical analysis were identified 2,226 differentially modulated proteins. Of these, 20 plasma membrane localized receptor like kinases (RLKs) with the increased abundance in response to MSP1. Moreover, activation of proteins related to the protein degradation and modification, calcium signaling, transcription factor, redox, and MAPK signaling were characterized. Taken together, our results indicated that potential PRR candidates involved in response to blast disease and thus suggesting the overview mechanism of the MSP1-induced signaling in rice leaves.
Project description:For gene expression profiling, we used immortalized human mammary epithelial cells (HMLE) to isolate a pure epithelial fraction of cells by positive selection for CD24 expression using Magnetic Activated Cell Sorting (=24hi). We independently isolated 3 mesenchymal subpopulations (msp1-3) from HMLE cells by collecting floating cells from cultured monolayer HMLE cells. See summary