Project description:Plasma membrane NADPH oxidases (NOXs) are major producers of reactive oxygen species (ROS) in plant cells under normal growth and stress conditions. Rice NOXs have multiple homologs but their functional mechanisms are largely unknown. We used microarrays to detail the global gene expression profiles in rice wild-type (WT, Dongjin) and a mutant osnox2 which loss the functions of OsNOX2 protein under drought and identified distinct classes of genes between the two type rice plants under both normal growth and drought stressed conditions.
Project description:To understand the transcriptional response of mammalian cells to plasma membrane stress we induced partial loss of plasmid membrane integrity by chemogenetic induction of pore forming proteins (Gasderimin and MLKL) or treatment with the detergent digitonin. We monitored transcription with mRNAseq in the immediate hours after plasma membrane damage and identified conserved transcriptional programs.
Project description:Plasma membrane NADPH oxidases (NOXs) are major producers of reactive oxygen species (ROS) in plant cells under normal growth and stress conditions. Rice NOXs have multiple homologs but their functional mechanisms are largely unknown. We used microarrays to detail the global gene expression profiles in rice wild-type (WT, Dongjin) and a mutant osnox2 which loss the functions of OsNOX2 protein under drought and identified distinct classes of genes between the two type rice plants under both normal growth and drought stressed conditions. The youngest fully expanded leaves from 2.5-month-old WT and osnox2 plants (three replicates each), grown under normal growth (soil moisture, 47.3%) and drought conditions (soil moisture, 8.5%), were used for RNA extraction and hybridization on Affymetrix microarrays. Control: normal growth condition; Drought: drought stress condition.
Project description:This project aims to analyze rice plasma membrane proteins related to resistance against rice blast infection. We extracted rice plasma membrane proteins before and after M.oryzae infection for 24h, and then used trypsin to digest and iTRAQ to label the peptides, HPLC-MS/MS was used to seperate and identify peptides. 1.1/0.909 fold change with p-value < 0.05 was used as threshold for differentially expressed proteins. 2,977 proteins were identified and 951 of which were found to be responsive to resistance against M. oryzae. Gene Ontology (GO) enrichment analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis and protein interaction network showed that plenty of proteins were involved in vesicle trafficking with obvious functional tendencies towards transport, vesicle-mediated transport, secretion, endocytosis and phagosome. 10 DEPs were validated at transcript level, and a SNARE protein named NPSN (novel plant-specific SNARE)13 actively responded to M. oryzae infection, and it contributed to rice blast resistance and mainly located at PM.
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:The coordination of pollen tube (PT) growth, guidance and timely growth arrest and rupture mediated by PT-pistil interaction is crucial for the PT to transport sperm cells into ovules for double fertilization. The plasma membrane (PM) represents an important interface for cell–cell interaction, and PM proteins of PTs are pioneers for mediating PT integrity and interaction with pistils. Thus, understanding the mechanisms underlying these events is important for proteomics. Using the efficient aqueous polymer two-phase system and alkali buffer treatment, we prepared high-purity PM from mature and germinated pollen of rice. We used iTRAQ quantitative proteomic methods and identified 1,121 PM-related proteins (PMrPs) (matched to 899 loci); 192 showed differential expression in the two pollen cell types, 119 up- and 73 down-regulated during germination. The PMrP and differentially expressed PMrP sets all showed a functional skew toward signal transduction, transporters, wall remodeling/metabolism and membrane trafficking. Their genomic loci had strong chromosome bias. We found 37 receptor-like kinases (RLKs) from 8 kinase subfamilies and 209 transporters involved in flux of diversified ions and metabolites. In combination with the rice pollen transcriptome data, we revealed that in general, the protein expression of these PMrPs disagreed with their mRNA expression, with inconsistent mRNA expression for 74% of differentially expressed PMrPs. This study, for the first time, identified genome-wide pollen PMrPs, and provided insights into the membrane profile of receptor-like kinases and transporters important for pollen tube growth and interaction with pistils. These pollen PMrPs and their mRNAs showed discordant expression. This work provides novel resource and knowledge to further dissect mechanisms by which pollen or the PT controls PMrP abundance and monitors interactions and ion and metabolite exchanges with female cells in rice.