Project description:To dissect the mechanisms that control and mediate cellular iron homeostasis, we conducted quantitative high-resolution iTRAQ proteomics and microarray-based transcriptomic profiling of iron-deficient Arabidopsis thaliana plants. Proteomic and transcriptomic profiling of Arabidopsis Col and RING DOMAIN LIGASE1 (RGLG1) and RING DOMAIN LIGASE2 (RGLG2) double mutation in response to iron deficiency were conducted. This integrative analysis provides a detailed catalog of post-transcriptionally regulated proteins and allows the concept of a chiefly transcriptionally regulated iron deficiency response to be revisited.
Project description:Purpose: plants exposed to multiple simultaneous adverse growth conditions trigger molecular responses that differ from the sum of those to individual stressors. Copper and iron are fundamental elements required for proper photosynthesis, energy production, DNA metabolism and hormone sensing, among all. Therefore, copper and iron deprivation limits plant yield. In natural environments, simultaneous deficiency to copper and iron can occur. As part of a multiple high-throughput study to identify combinatorial responses to both copper and iron deficiency, RNA-Seq profiling of Arabidopsis thaliana rosette leaves exposed to copper and/or iron deficiencies have been conducted. Methods: RNA-Seq libraries were prepared from total RNA of whole rosettes of 20-d-old plants treated for control conditions, copper deficiency, iron deficiency or simultaneous deficiency to both copper and iron for 10 d and sequenced using Illumina protocols. 2 independent plants were RNA-Seq-sequenced per treatment. Adaptor sequences were removed with Trimmomatic and the resulting reads mapped to the Arabidopsis genome (Araport11) with Tophat 2.1.1. Read counts and differential expression analysis were conducted with Cufflinks/Cuffdiff. Results: for RNA-Seq analysis a Tophat/Cuffdiff pipeline was designed. Each sample provided app. 9 million reads. After applying a cut-off of absolute log2(FC) ≥ 1 to controls and a FDR ≤ 0.05, copper deficiency led to 83 differentially expressed genes, followed by 1708 during iron deficiency, while the combinatorial treatment altered 2056 transcripts. Comparison of differential expressed genes among treatments indicated that double deficiency led to app. 45% rewiring of all detected transcriptional changes. Conclusions: our data support that combinatorial copper and iron deficiency treatments in plants triggers transcriptional responses that differ from those to single deficiencies.
Project description:Arabidopsis thaliana Col-0 plants and three other genotypes (ARR1 overexpressor, arr1-1 knockout, overexpressor of ARR1-SRDX fusion protein) were grown in liquid media (1/2 MS, 1 g/L sucrose, 0.5 g/L MES, pH 5.7) in a Percival AR-66L growth chamber at 24 oC, 16:8 h day:night cycle, and 100 µE light intensity until growth stage 1.0. Plants were then treated with 5 µM 6-Benzyladenine for 0, 15, and 120 min, harvested and frozen in liquid nitrogen for RNA extraction and subsequent processing for microarray hybridization.
Project description:In order to identify differentially expressed genes in developing seeds of Arabidopsis thaliana three different stages of seed development were analysed (9-10, 10-11 and 12-13 days after flower opening) for two Arabidopsis thaliana accessions, Col-0 and C24. For each stage and accession three biological replicates were analysed.
Project description:In order to identify differentially expressed genes in developing seeds of Arabidopsis thaliana three different stages of seed development were analysed (9-10, 10-11 and 12-13 days after flower opening) for two Arabidopsis thaliana accessions, Col-0 and C24. For each stage and accession three biological replicates were analysed.
Project description:Purpose: plants exposed to multiple simultaneous adverse growth conditions trigger molecular responses that differ from the sum of those to individual stressors. Copper and iron are fundamental elements required for proper photosynthesis, energy production, DNA metabolism and hormone sensing, among all. Therefore, copper and iron deprivation limits plant yield. In natural environments, simultaneous deficiency to copper and iron can occur. As part of a multiple high-throughput study to identify combinatorial responses to both copper and iron deficiency, proteomic profiling of Arabidopsis thaliana rosette leaves exposed to copper and/or iron deficiencies have been conducted.
Project description:Iron-sulfur (Fe-S) clusters play an essential role in plants as protein co-factors mediating diverse electron transfer reactions. Because they can react with oxygen to form reactive oxygen species (ROS) and cause cellular damage, the biogenesis of Fe-S clusters is highly regulated. A newly discovered group of 2Fe-2S proteins, termed NEET proteins, was recently proposed to coordinate iron-sulfur, iron, and ROS homeostasis in mammalian cells. Here we report that disrupting the function of AtNEET, the sole member of the NEET protein family in Arabidopsis thaliana, triggers a leaf-associated Fe- and Fe-S-deficiency responses, iron overload in chloroplasts (1.2-1.5 fold), chlorosis, structural damage to chloroplasts, and a high seedling mortality rate. Our findings suggest that disrupting AtNEET function disrupts the transfer of 2Fe-2S clusters from the chloroplastic 2Fe-2S biogenesis pathway to Fe-S proteins, and that uncoupling this process triggers a leaf-associated Fe-deficiency response that results in iron overload in chloroplasts and enhanced ROS accumulation
Project description:RNAseq transcriptome of anthers of Arabidopsis thaliana Columbia-0 grown under control (1/2 Hoagland) and Fe deficiency conditions.
