Transcription profiling of Nicotiana tabacum grown in different light conditions to test the reproducibility of a custom Affymetrix microarray for tobacco
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ABSTRACT: A study was carried out to compare gene expression levels between technical replicate samples in order to test the reproducibility of a custom Affymetrix microarray for Tobacco (Nicotiana tabacum).
Project description:Gene expression was measured in different tissues throughout the lifecycle of the Nicotiana tabacum plant to generate the Tobacco Expression Atlas (TobEA).
Project description:Gene expression was measured in leaves from dark treated tobacco plants to investigate the changes associated with dark induced senescence.
Project description:This RNA-seq was designed to help gain understanding on the genetic program behind phytochrome developmental time dependent control of leaf 3 (L3) cell proliferation and expansion phases, to ultimately regulate organ growth. Briefly, Arabidopsis thaliana Col-0 plants were grown under a light : dark (LD) 12 h : 12 h photoperiod, at a 100 µmoles/m2/s fluence rate and 21 °C of constant temperature. Plants were then exposed to either daily End of Day (EoD) far-red (FR, 40µmoles/m2/s of FR light (730nm) for 10 minutes) from day 6 and harvested L3 primordia or blade tissue at days 13, 16 and 20, or EoD FR from day 18, sampling at day 20. Plant kept under white light (WL) conditions were used as controls.
Project description:Although significant work has been undertaken regarding the response of model and crop plants to heat shock during the acclimatory phase, few studies have examined the steady state response to the mild heat stress encountered in temperate agriculture. In the present work we therefore exposed tuberising potato plants to mildly elevated temperatures (30/20C), day/night) for up to five weeks and compared tuber yield, physiological and biochemical responses, and leaf and tuber metabolomes and transcriptomes with plants grown under optimal conditions (22/16C). Growth at elevated temperature reduced tuber yield despite an increase in net foliar photosynthesis. This was associated with major shifts in leaf and tuber metabolite profiles, a significant decrease in leaf glutathione redox state and decreased starch synthesis in tubers. Furthermore, growth at elevated temperature had a profound impact on leaf and tuber transcript expression with large numbers of transcripts displaying a rhythmic oscillation at the higher growth temperature. RT-PCR revealed perturbation in the expression of circadian clock transcripts including StSP6A, previously identified as a tuberisation signal. Our data indicate that potato plants grown at moderately elevated temperatures do not exhibit classic symptoms of abiotic stress but that tuber development responds via a diversity of biochemical and molecular signals. In this submission we are looking at gene expression changes with respect to both temperature and time, every 4h over a 24h period whereby diurnal changes may be apparent.
Project description:Maternal environment is an important regultor of seed dormancy, but the mechanisms underlying the process are poorly understood. We have found that genes in the circadian clock control dormancy, in part through their regulation of the canonical photoperiod pathway known from research into flowering time control. In this experiment we compare the affects of altering seed maturation temperature or maternal photoperiod on dry seed transcriptomes, and the photoperiod-insenstive ft-1 mutant to wt type Ler. In this way we are identifying gene expression programmes which result from the seed's response to maternal environmental experience. Keywords: Expression profilling by array 12 samples were used in this experiment
Project description:To systematically investigate viral sRNA production and sRNA-target interaction, we sequenced sRNAs from Tobacco Rattle Virus (TRV)-infected Nicotiana benthamiana at an early (1 week post infection) and late time point (3 weeks post infection). The N. benthamiana 16c plants were infected with TGS-inducing viruses (TRV-35S and TRV-35-2M) and PTGS-inducing viruses (TRV-GFP and TRV-GFP-2M), respectively. TRV-35S is a recombinant TRV containing a 120 nt segment of the 35S promoter. Its derivative, TRV-35S-2M, carrying single nucleotide substitutions (SNS) at every 10 nt within the 120 nt 35S target segment. Same strategy was used to create recombinant TRV-GFP and TRV-GFP-2M targeting GFP coding sequence. According to SNS content, sRNAs from TRV-35S-2M/TRV-GFP-2M infected plants can be separated to yield primary (containing SNSs) and secondary sRNAs (lacking SNSs). Wild Type TRV was used along as viral infection control. Libraries were indexed during PCR amplification (16 cycles) according to the Illumina protocol. See individual sample information for specific index primers used.
Project description:Yield losses as a result of abiotic stress factors present a significant challenge for the future of global food production. While breeding technologies provide potential to combat negative stress-mediated outcomes over time, interventions which act to prime plant tolerance to stress, via the use of phytohormone-based elicitors for example, could act as a valuable tool for crop protection. However, the translation of fundamental biology into functioning solution is often constrained by knowledge-gaps. Photosynthetic and transcriptomic responses were characterised in young tomato (Solanum lycopersicum) seedlings in response to pre-treatment with a new plant health activator technology, M-bM-^@M-^XAlethea,M-bM-^@M-^Y followed by a subsequent 100 mM salinity stress. Salinity treatment led to a maximal 47% reduction in net photosynthetic rate 8 d following NaCl treatment. In Alethea pre-treated seedlings, sensitivity to salinity stress was markedly reduced during the experimental period. Microarray analysis of leaf transcriptional responses showed that while salinity stress and Alethea individually impacted on largely non-overlapping, distinct groups of genes, Alethea pre-treatment substantially modified the response to salinity. Alethea affected the expression of genes related to biotic stress, ethylene signalling, cell wall synthesis, redox signalling and photosynthetic processes. Since Alethea had clear effects on photosynthesis/chloroplastic function at the physiological and molecular levels, we also investigated the ability of Alethea to protect plants against methyl viologen, a potent generator of oxidative stress in chloroplasts. Alethea pre-treatment produced dramatic reductions in visible foliar necrosis caused by methyl viologen compared with non-primed controls. Tomato leaves from four groups of treated plants were harvested at a single point following treatment for RNA extraction and hybridization on Affymetrix microarrays. Plants came from two initial groups, pre-treated either with water (control) or the Alethea product. Equal numbers of plants from within these groups were subsequently treated either with water (control) or salinity (100 mM NaCl), generating 4 main experimental groups in a 2x2 factorial design. Samples were collected from 3 biological replicate experiments, and each sample included leaves pooled from 3 individual plants from each treatment group.
Project description:To investigate how high temperature affects sRNA production during virus induced gene silencing (VIGS), we sequenced sRNAs from Tobacco Rattle Virus (TRV)-infected Nicotiana benthamiana kept at 29°C at an early (1 week post infection) and a late time point (3 weeks post infection). To compare sRNA production between virus induced transcriptional gene silencing (ViTGS) and virus induced post-translational gene silencing (ViPTGS) at 29°C, the N. benthamiana 16c plants were infected with TGS-inducing viruses (TRV-35S and TRV-35-2M) and PTGS-inducing viruses (TRV-GFP and TRV-GFP-2M). TRV-35S is a recombinant TRV containing a 120 nt segment of the 35S promoter. Its derivative, TRV-35S-2M, carrying single nucleotide substitutions (SNS) at every 10 nt within the 120 nt 35S target segment. Same strategy was used to create recombinant TRV-GFP and TRV-GFP-2M targeting GFP coding sequence. According to SNS content, sRNAs from TRV-35S-2M/TRV-GFP-2M infected plants can be separated to yield primary (containing SNSs) and secondary sRNAs (lacking SNSs). Wild Type TRV was used along as viral infection control. Libraries were indexed during PCR amplification (16 cycles) according to the Illumina protocol. See individual sample information for specific index primers used.
Project description:Exon array analysis was performed using retinae from three transgenic mouse models, coneless (cl), rodless (rd/rd) and rodless/coneless (rd/rd cl) compared to wildtype (non-rd) allowing gene expression in the rod and cone photoreceptors to be examined.