Project description:We performed mass-spectrometry analysis of proteins co-purifying with Whi5, a predicted repressor of the MBF transcriptional complex. Whi5 was tagged with GFP at its C-terminal end and expressed from its endogenous promoter at its chromosomal locus. Whi5-GFP was immunoprecipitated from cells growing in MM and after 16 hours in MM-N, and co-purifying proteins were analysed by mass-spectrometry. An untagged wild-type strain was used as a control.
Project description:To identify novel miRNA and NAT-siRNAs that are associated with abiotic stresses in sorghum, we generated small RNA sequences from sorghum seedlings that grew under control and under dought, salt, and cold stress treatments. sequencing of small RNAs in sorghum under control, drought, salt, and cold stress conditions.
Project description:To identify novel miRNA and NAT-siRNAs that are associated with abiotic stresses in maize, we generated small RNA sequences from maize seedlings that grew under control and under dought, salt, and cold stress treatments. Sequencing of small RNAs in maize under control, drought, salt, and cold stress conditions.
Project description:To explore the possible neddylation sites, GFP-IRF7 and His-NEDD8 were co-overexpressed in HEK293 cell line and GFP was immunoprecipitated, followed by SDS-PAGE and silver staining. The band detected was subjected to LC–MS/MS.
Project description:To identify novel miRNA and NAT-siRNAs that are associated with abiotic stresses in Arabidopsis, we generated small RNA sequences from adult Arabidopsis plants under control and under dought, salt, and cold stress treatments. Over 23 million reads were generated. Sequencing of small RNAs in Arabidopsis under control, drought, salt, and cold stress conditions.
Project description:To identify novel miRNA and NAT-siRNAs that are associated with abiotic stresses in rice, we generated small RNA sequences from inflorescences from rice under control and under dought, salt, and cold stress treatments. Over 30 million reads were generated. sequencing of small RNAs in rice under control, drought, salt, and cold stress conditions.
Project description:We carried out immunoprecipitation-mass spectrometry (IP-MS) analysis to identify the protein components of PGL granules in heat-stressed embryos. GFP::PGL-3 proteins were immunoprecipitated from extracts of embryos grown at 26 oC and the interacting proteins were identified by LC-MS/MS. Proteins involved in translation and RNA control factors (e.g. RNA binding proteins, ribonucleases, RNA helicases, proteins involved in siRNA-mediated mRNA turnover) were enriched in the GFP::PGL-3 co-immunoprecipitants
Project description:The growth and fruit quality of grapevine are widely affected by abnormal climatic conditions such as water deficit. But how grapevine responds to drought stress is still largely unknown. Here we found that VaNAC26, a member of NAC transcription factor family, was up-regulated dramatically during cold, drought and salinity treatments in Vitis amurensis, a cold and drought-hardiness wild Vitis species. Ectopic overexpression of VaNAC26 enhanced the drought and salt tolerances in transgenic Arabidopsis. Higher activities of antioxidant enzymes and the lower concentration of H2O2 and O2- were found in VaNAC26-OE lines than in wild type plants under drought stress. These results indicate that the reactive oxygen species (ROS) scavenging was enhanced by VaNAC26 in transgenic lines. Microarray based transcriptome analysis reveals that genes related to jasmonic acid (JA) synthesis and signaling were up-regulated in VaNAC26-OE lines under both normal and drought conditions. VaNAC26 showed a specific binding ability on NACRS motif, which was broadly existent in the promoter regions of up-regulated genes in transgenic lines. Endogenous JA content was found increased obviously in VaNAC26-OE-2/3 lines. Our data suggests that VaNAC26 responds to abiotic stresses and may enhance the drought tolerance by transcriptional regulation of JA synthesis in Arabidopsis.
Project description:This dataset describes a proteomic analysis of proteins co-immunoprecipitated with functional GFP-tagged PILS2, PILS3, and PILS6 fusion proteins in Arabidopsis thaliana roots. PILS (PIN-LIKES) proteins are auxin transport facilitators that localize to the endoplasmic reticulum (ER). To identify proteins that associate with PILS2, PILS3, and PILS6 under physiological conditions, we performed GFP-based immunoprecipitation (IP) followed by mass spectrometry (MS). Protein extracts were prepared from root tissues of Arabidopsis Col-0 (wild-type) and PILS2-, PILS3-, and PILS6-overexpressing transgenic lines expressing N- or C-terminal GFP fusion proteins. Plant material was ground in liquid nitrogen, and total proteins were extracted using aTris Buffer containing 1% CHAPS, 20 mM Tris-HCl (pH 7.5), 10 mM MgCl₂, 15 mM EGTA, 75 mM NaCl, 1 mM DTT, 0.1% Tween-20, and protease inhibitors. Extracts were centrifuged, and the supernatants were quantified using the Bradford assay. 2mg of protein were subjected to immunoprecipitation using anti-GFP magnetic beads, Col-0 (non-tagged) extracts were used as negative controls. Co-immunoprecipitated proteins were then processed and analyzed by mass spectrometry.
Project description:Our study showed that the CHD3 protein PKL plays a role in the regulation of cold stress response likely via the regulation of chlorophyll accumulation under low temperature conditions. Our results suggest that PKL may regulate cold response partly via a CBF3-mediated pathway. Besides, our results reveal that the PKL gene is also involved in the regulation of drought and salt stress resistance.