Profiling primary and secondary targets of ARF7 and ARF19 in Arabidopsis thaliana roots
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ABSTRACT: Arabidopsis thaliana AF7/ARF19 double knockout with ARF7 reintroduced under its own promotor with a glucocorticoid receptor added were treated with Auxin, Dexamethazone and cycloheximide to determine primary and secondary ARF7 auxin sensitive downstream targets
Project description:This research reports the analysis of sRNAs in 14 and 7 inbred lines from a breeding population. We analyzed the contribution of sRNAs to the formation of heterosis via integrative association analysis with field data of 98 hybrids generated from the set of inbred lines. Our results indicate a contribution of sRNAs to heterosis. We were able to identify different sets of sRNAs associated with heterosis with distinct length and genome distribution patterns. Analysis of sRNA contribution to the formation of heterosis in maize by an association study in a breeding population.
Project description:We developed a method to synchronize the induction of lateral roots in primary and adventitious roots of Zea mays, and used it to perform a genome-wide transcriptome analysis of the pericycle cells in front of the phloem poles during lateral root initiation. Lateral roots were induced in primary and adventitious roots of Maize. For the primary root, plants were germinated and grown 64 hours in NPA 50 µM, and then transfered to NAA 50 µM. For the adventitious roots, plants were germinated and grown in water for 6 days, then tranfered 4 days in NPA 25 µM, and finally transfered to NAA 25 µM. For all these roots, pericycle cells located in front of the phloem poles in segments of roots located between 5 and 10 mm distance from the root tip were isolated using laser capture microdissection after cryosection. Material was sampled at 0 hours (NPA) and after 2, 3 and 4 hours of NAA treatment, for both the primary and adventitious roots and also after 6 hours and 9 hours of NAA treatment for the adventitious roots.
Project description:Abscisic acid (ABA) is a plant hormone that is important in responding to various environmental stresses. Using an ABA auxotroph in Arabidopsis (aba2-2) as the plant material, we would like to identify early downstream targets of transcription in response to a small dose of ABA, 1 uM. We also added cycloheximide to preferentially obtain immediate targets of ABA addition. We believe that using a sensitized background of an ABA auxotroph would yield a set of genes that are very closely regulated by ABA at the transcriptional level. This data set will be used for network analysis of ABA signaling.
Project description:ABI3 is a key regulator of seed development in Arabidopsis and other plants.To identify genes regulated by ABI3 we performed array based transcriptome analysis of Dexamethasone inducible ABI3 transgenic seedlings. ABI3 mostly in concert with abscisic acid (ABA) was found to activate genes specifically expressed during the maturation phase of seed development. Two weeks old wildtype and 35S::ABI3::GR seedlings were treated with ABA, Dexamethasone (DEX) and ABA plus Dexamethasone for 4 h. Two biological independent experiments were performed. Wildtype was induced to control for DEX-induction of genes.
Project description:TCP (TEOSINTE BRANCHED1/CYCLOIDEA/PCF1) transcription factors control developmental processes in plants. We identified direct target genes of the Arabidopsis class I TCP20 protein in leaf development based on a glucocorticoid receptor induction assay and genome-wide expression studies. For this, we tagged TCP20 with a glucucorticoid receptor (GR) domain and transformed the resulting TCP20-GR construct into tcp20 knockout plants. Induction of these and wild type controls was done with Dexamethasone and Cycloheximide. Plants were harvested 8 h after induction, uninduced plants were taken as control. In sum, 8 samples, each pools of 30 seedlings. Wild type and TCP20-GR plants at induction times t=0 and t=8, in two biological replicates.
Project description:Direct target genes of VND7 were explored with inducible expression system using glucocorticoid receptor (GR). Transgenic plants expressing 35S:VND7-VP16-GR were treated with dexamethazone (DEX) and/or protein synthesis inhibitor cycloheximide (CHX). A number of genes related to the formation of vascular vessel was induced by DEX even in the presence of CHX. Total RNAs of the transgenic plants expressing 35S:VND7-VP16-GR treated with DEX plus CHX and those treated with CHX only were compared. As a control experiment, transgenic plants harboring empty vector were treated similarly and the total RNAs were compared similarly to identify genes merely induced by DEX treatment itself.
Project description:Pro35SLBD16:GR or Pro35SLBD18:GR transgenic seedlings that overexpress LBD16 or LBD18 fused to glucocorticoidsteroid hormone binding domain(GR) under CaMV35S promoter were grown for 12 days under long-day conditions (16h light/ 8h dark).
Project description:In plants, formation of functional stomatal guard cells is a highly regulated event so that this cell differentiation model constitutes an interesting genetic system to explore cell differentiation in response to cell cycle controllers or cell fate determinants such as transcription factors. The latest acting bHLH in the stomatal lineage circuit is FAMA that appears to be absolutely required to promote stomatal development by controlling the GMC to GC transition. In order for FAMA to trigger guard cell development, it probably initiates a gene activation cascade leading to cell differentiation and cessation of cell division. To identify and characterize FAMA inducible genes, transcriptome analysis using the Affymetrix ATH1 micro-array chip was carried out on inducible FAMA gain of function plants at 4hrs and 48 hrs post-induction. Transgenic plants with an estrogen-inducible FAMA expression cassette were used in a timecourse experiment to identify genes that were differentially regulated by FAMA. Two time points were analyzed, 4 hours and 48 hours post-induction. Seedlings were grown on vertically oriented plates in a 22 °C incubator under 24 hours light for 5 days. Plants were then transfered with forcepts to plates containing MS + Sucrose + 10 µM estrogen (or new MS + sucrose plates for controls). After 4h or 48h on new plates, samples were collected, frozen in liquid nitrogen and stored at â??80 degree until enough material was obtained to do all RNA extractions in the same week. The Rneasy Plant Mini Kit (QIAGEN) for RNA isolation, and samples were labelled with standard kits, etc (get info) and hybridized to Affymetrix Ath1 array chips. 15 samples.
Project description:Defining the transcriptome response to the brief induction of Golden2-like (GLK) gene expression in a glk mutant background. Expression of either GLK1 or GLK2 from an inducible promoter by the application of dexamethasone. Changes to transcriptome four hours after induction are compared with uninduced controls.