Project description:Members of the PIF quartet (PIFq) (PIF1, PIF3, PIF4, and PIF5) collectively contribute to induce growth in Arabidopsis seedlings under short day (SD) conditions, specifically promoting elongation at dawn. Their action involves the direct regulation of growth-related and hormone-associated genes. The objective of this genomic analysis was to define a comprehensive set of SD-regulated genes at dawn compared to free-running (LL) conditions in the wild type, followed by identification of those SD-regulated genes whose expression depends on the presence of PIFq. By using this sequential strategy, our aim was to identify PIF/SD-coregulated genes.
Project description:Transcriptional profiling of dark-grown Arabidopsis seedlings comparing SCR:PIF1/pifQ transgenic plant with pif1pif3pif4pif5 quadruple mutant (pifQ). Seedlings were grown under dark condition for 2.5 days. Goal was to determine the effects of endodermal PIF1 in dark-grown seedlings.
Project description:Transcriptional profiling of dark-grown Arabidopsis seedlings comparing SCR:PIF1/pifQ transgenic plant with pif1pif3pif4pif5 quadruple mutant (pifQ). Seedlings were grown under dark condition for 2.5 days. Goal was to determine the effects of endodermal PIF1 in dark-grown seedlings. Two-condition experiment, SCR:PIF1/pifQ vs. pifQ Biological replicates: 3 pifQ replicates, 3 SCR:PIF1 replicates.
Project description:Transcriptional profiling of 60h-old Arabidopsis whole seedlings comparing control Col-0 wild-type plants with pifQ mutant plants The expression profile of dark-grown pifQ mutant shows similar pattern of Rc-grown Col-0 wild-type Keywords: Genetic modification
Project description:The experiment was designed to get high throughput mRNA data (for gene expression and alternative splicing analysis) of A. thaliana 13-14 day old plants, over the course of two days after 24h of transfer to free running (circadian) conditions. Briefly, plants were grown in MS plates (30 plants per plate / 2 plates per timepoint) for 11 days in LD 12 h : 12 h conditions (50umoles /m2 /s) and 22C. On day 12 plants were transferred to LL conditions. Sampling was performed at 4 h intervals from 24 h after transfer to LL till 68 h. Samples were flash frozen in liquid nitrogen and total RNA was extracted using the Trizol method. RNA quality was chequed by gel and libraries were constructed using the TruSeq RNA Library Prep Kit v2. QC was performed on an Agilent BioAnalyzer 2100. Libraries were then pooled and sequenced on an Ilumina HiSeq 2500 by INDEAR, Rosario, Argentina.
Project description:Plants respond to changes in the red:far red ratio (R:FR) of incident light. A reduction in this ratio (increase in FR) results in the Shade Avoidance Response (SAR) with associated changes in gene expression. The Phyotchrome-Interacting Factors (PIFs) are bHLH transcription factors known to be involved in the SAR. An analysis of changes in gene expression in WT and quadruple pif1pif3pif4pif5 (pifq; Leivar et al., 2008 (PMID 19920208)) mutant seedlings in response to an increase in FR should identify primary targets of PIF signaling. We used microarrays to examine the SAR in WT (Columbia) and pifq mutant Arabidopsis seedlings. Arabidopsis WT and pifq mutant seeds were plated on GM medium without sucrose at room temperature. During this procedure, the seeds were routinely exposed to white light (WL) for a total of 1.5 hours after imbibition. Seeds were then stratified for 5 days at 4ºC in darkness, and then grown in WL (19 umol/m2/s, R/FR ratio of 6.48) for 2 days at 21°C (WL0 samples). Two-day-old WL-grown seedlings were then maintained in the same fluence rate of WL supplemented with far-red light (WL-FR, R/FR ratio of 0.006) for 1 (FR1), 3 (FR3) or 24 (FR24) hours before harvesting. Control seedlings were also maintained in parallel in the same fluence rate of WL for 24h (WL24) before harvesting. Three different biological replicates of each treatment were grown separately and extracted, processed, and analyzed independently.
Project description:For expression profiling analyses of early stages of tuber induction, plants of Solanum tuberosum ssp andigena (7540) were used. This wild subspecies is strictly dependent on photoperiod for tuberisation, such that short days (SD) inductive conditions are required in order to trigger tuber induction in the stolons. Andigena plants were grown in the greenhouse under LD non-inductive conditions until a 10-leaf stage. They were subsequently transferred to inductive SD conditions (8 h light/16 h dark), and sampled at 0, 2, 4, 6 and 8 days after transfer to SDs. Tuber swelling was visible approximately 6-8 days after transfer to inductive conditions. The apical region of the stolons (2 cm) was collected one hour before the beginning of the light period.
Project description:Plants of Solanum tuberosum ssp andigena, which tuberize only under SD conditions, were grown in growth chambers under non-inductive LD conditions and half of the plants were transferred to inductive SD conditions for 1 or 15 days, whilst the rest of the plants were kept as controls in LD. Transgenic potato plants with reduced phyB levels, which tuberize irrespective of photoperiod, were also grown in LD conditions. All the plants, irrespective of treatment, were harvested 14 hours after the beggining of the photoperiod (i.e. immediately before dusk for plants on LD, and 8 hours after dusk for the plants on SD). A similar experimental protocol was used with plants of N. tabacum cv Hicks that flower at the same time irrespective of photoperiod and the isogenic line N. tabacum cv Hicks carrying the Maryland Mammoth mutant allele, which flower only on SD (i.e. the plants were grown under LD for 55 or 41 days in LD and then transferred for 1 or 15 days to SD conditions, respectively, whilst control plants were grown 56 days in LD). Finally, Nicotiana silvestris plants that flower only on LD were grown in non-inductive SD and then half of the plants were transferred to inductive LD conditions for 1 or 15 days. In all cases we harvested only the leaves, the organ in which daylength perception takes place and photoperiodic responses are initiated. SD conditions in this experiments were 8 hours of light/ 16 hours of darkness, 160 uEinstein, 22 °C. LD conditions were 16 hours of light/ 8 hours of darkness, 80 uEinstein, 22 °C. RNA was extracted with TRIZOL. Keywords: Direct comparison / Balanced Block Design