Project description:Arabidopsis seedlings were treated with 1 uM RALF peptide for 5 min to examine rapid protein phosphorylation changes. Phosphopeptides from the plasma membrane were identified and quantified using Orbitrap mass spectrometer.
Project description:Hrip1 was isolated from Alternaria tenuissima and was found to induce systematic acquired resistance in tobacco. In rice plants, it was found to confer immunity against rice blast fungi. To understand the genomic signature of Hrip1-induced immunity, rice seedlings were treated with 30 nm Hrip1 protein and triplicate leaf samples were subjected to sequencing on the BIGSEQ500 platform. Two groups of rice seedlings were grown to the three-leaf stage. The experiment group was treated with 30nm Hrip1 protein while the control plants were treated with Tris-HCl buffer. Triplicate samples from both setups were taken at 6, 12, 24, and 48 hr after treatment. Total RNA was extracted and reverse transcribed in first strand cDNA as input material.
Project description:An auxin-binding protein (Abp57) was previously isolated from rice and known to activate plasma membrane proton ATPase. The Abp57 function was characterised by overexpression in the rice and Arabidopsis. The transgene expression was driven by constitutive promoter, CaMV35S. Results from physiological experiments showed that the transgenic lines were tolerant to drought and salinity stress. The global effects of Abp57 overexpression in rice were studied using microarray platform from Affymetrix.
Project description:A tandem mass tag (TMT)-based comparative peptidomics analysis of rice seedlings under salt stress was conducted. Rice seedlings were exposed to 50 and 150 mM NaCl for 24 and 72 h, respectively, and the root and shoot tissues of different treatment groups were collected separately for the peptidomic analysis.
Project description:Rice blast disease is a major threat to rice production worldwide, but the mechanisms underlying rice resistance to the causal agent Magnaporthe oryzae remain elusive. In this whole-genome transcriptome study of rice early defense response to M. oryzae, we applied Affymetrix Rice Genome Genechip to compare the compatible and incompatible rice-M. oryzae interactions in 24 hours post-inoculation. Leaf samples were harvested from three biological replicates of fungal- and mock-inoculated seedlings at 24 hours post-inoculation, from which RNA were extracted and analyzed with Genechip Rice Genome Array.
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:Calcium is an essential macronutrient and plant requires it in large amounts for normal growth and development. This ion participated in innumerable processes and affects nearly all aspects of plant growth and development such as signal transduction, metabolism of lipids, proteins, and carbohydrates, cell growth, cell wall and membrane stabilization. We used whole genome microarrays to determine the transcriptomic profile of rice seedlings exposed to short-term and long term Ca2+ deficiency followed by Ca2+ resupply. Calcium treated seedlings were used for for RNA extraction and hybridization on Agilent microarray platform. Three biological replicates of each sample were used for microarray analysis. We wanted to know the altered expression patterns of calcium-responsive genes majorly involved in metabolic processes, signal transdction pathways, transcriptional regulation, and transport of multiple molecules including Ca2+. Seeds of indica rice were surface sterilized and grown hydroponically in rice growing medium. Plants were grown in the rice growth room under the condition of 16 h light/8 h dark (28M-BM-0C) photoperiod with 70 % humidity. After 5 days of normal growth, some of the seedlings were transferred to nutrient solution lacking CaCl2 (-Ca2+). After 5 and 14 days, calcium (0.798 mM CaCl2) was resupplied to the plants grown in -Ca2+ medium for 6 h.