Project description:Cycling Dof Factor (CDF) transcription factors have been implicated in different aspects of plant growth and development. In Arabidopsis and tomato, two members of this family CDF1 and CDF3 have recently been associated with the regulation of primary metabolism and abiotic stress responses, but their roles in crop production under open field conditions remain unknown. In this study, we compared growth and tuber yield and composition of wild-type (WT) potato plants and plants ectopically expressing the CDF1/3 genes from Arabidopsis under the control of the 35S promoter cultured under growth chamber and open field conditions. In growth chambers, the 35S::AtCDF1 and 35S::AtCDF3 potato plants showed higher than WT biomass production and tuber yield. Under field conditions, 35S::AtCDF1 plants showed significant increases in tuber yield with a significant increase in tuber weight, whereas 35S::AtCDF3 plants showed no significant changes in yield, but produced more than WT number of tubers. Metabolomic analysis revealed that tubers of 35S::AtCDF1 plants cultured under open field conditions accumulated higher levels of glucose, starch and amino acids than WT tubers. Comparative proteomic analysis of tubers of 35S::AtCDF1 and control plants cultured under open field revealed that these changes can be accounted for by changes in the expression of proteins involved in energy production and different aspects of C and N metabolism. Results from this study advance our collective understanding of the role of CDFs and are of interest to improve yield and breeding of crop plants
Project description:AtACINUS protein is involved in regulation of alternative transcription and splicing(AS). Identifying interaction partners and protein complex compositions for AtACINUS can produce valuable information on the mechanisms by which they regulate transcription and AS, as well as post-translational modifications on AtACINUS. A homozygous 35S::AtACINUS-GFP/acinus-2 plant was selected for similar protein expression level to the endogenous AtACINUS protein of wild-type plants using a native α-AtACINUS antibody. We isolated putative AtACINUS interaction partners from young Arabidopsis seedlings using the native α-AtACINUS antibody. Plants expressing TAP-GFP under 35S promoter were used as controls.
Project description:AtACINUS protein is involved in regulation of alternative transcription and splicing(AS). Identifying interaction partners and protein complex compositions for AtACINUS can produce valuable information on the mechanisms by which they regulate transcription and AS, as well as post-translational modifications on AtACINUS. A homozygous 35S::AtACINUS-GFP/acinus-2 plant was selected for similar protein expression level to the endogenous AtACINUS protein of wild-type plants using a native α-AtACINUS antibody. We isolated putative AtACINUS interaction partners from young Arabidopsis seedlings using a modified LaG16LaG2 nanobody. Plants expressing TAP-GFP under 35S promoter were used as controls.
Project description:Transcriptional profiling of Arabidopsis thaliana, comparing control wild-type (ecotype Wassilewskija, Ws) leaves with leaves from transgenic plants overexpressing the transcription factor RAP2.6L under control of the cauliflower mosaic virus 35S promoter (RAP2.6L-OX; this line was originally described in Krishnaswamy et al (2010)).
Project description:To study the genes regulated by transcription factor SQUAMOSA PROMOTER BINDING PROTEIN-LIKE7 (SPL7) in Arabidopsis thaliana, we conducted chromatin immunoprecipitation-based sequencing (ChIP-seq) in 35S:FALG-SPL7 transgenic plant and spl7 mutant, and genome-wide sequencing-based transcript profiling of Arabidopsis thaliana wild-type plants and of spl7 mutant under MS medium and MS supplement with 5µM CuSO4. These experiments led to the identification of genes that are direct target of SPL7 and genes differentially expressed in an SPL7-dependent or Cu-dependent manner. This study provides a framework for the identification of SPL7 regulated genes towards characterization of SPL7 in copper homeostasis.
Project description:A novel cold-inducible GSK3/Shaggy-like kinase cDNA (TaSK5) was isolated from winter wheat by a macroarray-based differential screening approach. Sequence analysis of TaSK5 revealed high similarity to Arabidopsis subgroup I GSK3/Shaggy-like kinases ASK-alpha, ASK-gamma and ASK-epsilon. Transgenic Arabidopsis plants overexpressing TaSK5 cDNA under the control of CaMV 35S promoter showed enhanced tolerance to salt and drought stresses. In contrast, the tolerance of the transgenic plants to freezing stress was not altered. To identify genes which are differentially regulated in the 35S:TaSK5 over-expressing Arabidopsis plants under non-stress conditions, we compared the genome-wide expression profiles of Col-0 and plants over-expressing TaSK5 using DNA microarrays. Sixty seven genes were found to be expressed at least 2-fold more strongly in 35S:TaSK5 plants than in Col-0, and 17 genes were found to be expressed at least 2-fold more strongly in Col-0 than in 35S:TaSK5 plants. Most of the TaSK5 up-regulated genes were also induced by abiotic stresses, including cold, salt and drought. These results support the involvement of TaSK5 in abiotic stress signal transduction. Keywords: transgenic vs wt Col.-0 comparison
Project description:A novel cold-inducible GSK3/Shaggy-like kinase cDNA (TaSK5) was isolated from winter wheat by a macroarray-based differential screening approach. Sequence analysis of TaSK5 revealed high similarity to Arabidopsis subgroup I GSK3/Shaggy-like kinases ASK-alpha, ASK-gamma and ASK-epsilon. Transgenic Arabidopsis plants overexpressing TaSK5 cDNA under the control of CaMV 35S promoter showed enhanced tolerance to salt and drought stresses. In contrast, the tolerance of the transgenic plants to freezing stress was not altered. To identify genes which are differentially regulated in the 35S:TaSK5 over-expressing Arabidopsis plants under non-stress conditions, we compared the genome-wide expression profiles of Col-0 and plants over-expressing TaSK5 using DNA microarrays. Sixty seven genes were found to be expressed at least 2-fold more strongly in 35S:TaSK5 plants than in Col-0, and 17 genes were found to be expressed at least 2-fold more strongly in Col-0 than in 35S:TaSK5 plants. Most of the TaSK5 up-regulated genes were also induced by abiotic stresses, including cold, salt and drought. These results support the involvement of TaSK5 in abiotic stress signal transduction. Keywords: transgenic vs wt Col.-0 comparison Total RNA was extracted from rosette leaves of two independent 3-week-old T1 Arabidopsis Col-0 plants over-expressing TaSK5 driven by 35S promoter and the whole transcriptome was compared with that of wild-type plant. Sample pairs in two independent transformants (OX7 and OX17), referred as biological replicates, were analyzed by the two-color method. Dye-swapped hybridizations were performed in every replicate.
Project description:To identify possible mitochondrial DNA binding targets of SHOT1/MTERF18 (AT3G60400), C-terminal GFP fusion constructs under either a constitutive 35S promoter (35S::SHOT1-GFP) or the SHOT1 native promoter (SHOT1p::SHOT1-GFP) were made. The transgenes were introduced into the shot1-2 mutant background (Kim et al., 2012, Plant Cell) and verified to be functional based on recovery of shot1 from growth defects. As a control, a transgenic line harboring mitochondria-targeted GFP (Mito-GFP) under the 35S promoter was used. All enriched peaks in SHOT1-GFP samples are located upstream of different tRNA genes: trnS, trnM, trnG and trnF