Transcription profiling by array of transgenic tobacco plants expressing begomoviral AC2 RNA silencing suppressor to compare transcriptional changes between transgenic and wild type plants
ABSTRACT: Transgenic tobacco plants expressing begomoviral AC2 RNA silencing suppressor were used to compare transcriptional changes in the transcriptome between transgenic and wild type tobacco plants. Transcriptional analysis using Agilent 4x44k tobacco array was performed of six week-old leaves and 3-5 month-old flowers taken from the same plants as leaf samples.
BACKGROUND: RNA-silencing is a conserved gene regulation and surveillance machinery, which in plants, is also used as major defence mechanism against viruses. Various virus-specific dsRNA structures are recognized by the silencing machinery leading to degradation of the viral RNAs or, as in case of begomoviruses, to methylation of their DNA genomes. Viruses produce specific RNA silencing suppressor (RSS) proteins to prevent these host defence mechanisms, and as these interfere with the silencing ...[more]
Project description:Agilent 4x44k tobacco micro array of wild type tobacco, empty vector control, and HC-Pro transgenic tobacco plants. Both 1-month old leaves and flowers were analyzed. Three biological replicates were performed of each sample.
Project description:We have used a microarray approach to study the effects of the Potato Virus X Potexvirus (PVX)-specific P25 VRS protein on the transcript profile of tobacco plants, when expressed as a transgene in these plants.
Project description:Agilent 4x44k tobacco micro array of wild type tobacco (WT) and whole tobacco mosaic virus (TMV) containing transgenic tobacco plants. The transgenic plants before resistance break (BRB-6 weeks), after resistance break (ARB-8 weeks) and wild type tobacco plants infected with TMV (TMVi-9weeks) leaves were analyzed. Three biological replicates were performed for each sample.
Project description:Transcriptome analysis in tobacco mutant plants using tomato Genechip Genome array Tobacco (Nicotiana tabacum cv. Petit Havanna) psaA and psbA deletion mutants were constructed through a targeted deletion of 767 and 1152 nucleotides of coding regions, respectively with two gene cassettes: psbAproR:uidA:psbterR and rrnR:aadA:rbcLterR coding for GUS reporter and spectinomycin selectable marker genes, respectively. Standard established procedures were followed for chloroplast transformation to generate the psaA and psbA deletion mutants based on the homologous recombination. Gene expression profiles in psaA and psbA tobacco mutant plants were analyzed using tomato Genechip Genome array to study the global changes in the expression of genome. Total RNA was isolated from psaA and psbA tobacco mutant plants along with the wild type plants. Biotin labeled cRNA was hybridized on tomato GeneChip Genome Array following the Affymetrix protocols. Two independent biological replicates were maintained.
Project description:Transcriptome profiling of three developmental stages of immature male gametophyte intobacco (Nicotiana tabacum) Total RNA isolated from tobacco microspores and early and late bicellular pollen was hybridised on Agilent Tobacco Gene Expression Microarray 4x44K in two biological replicates per sample
Project description:We compared the transcriptional profiles of female adult whiteflies of B. tabaci Middle East-Asia Minor 1 feeding on TYLCCNV-free and TYLCCNV-infected tobacco plants using the next-generation sequencing technique. Culture of B (MEAM1 cryptic species) whitefly was maintained on cotton plants. One thousand of newly emerged adults of whitefly on cotton were released onto the leaves of healthy and viruliferous tobacco plants. After 72 h of oviposition, all the adult whiteflies were discarded, and the progeny allowed to develop to adults. The cultures of MEAM1 on tobacco plants were maintained in climate chambers at 27 ± 1°C, a photoperiod of 14 h light/10 h darkness and 70 ± 10% relative humidity. Approximately 1,000 female adult whiteflies newly emerged from mock-inoculated tobacco plants and 1,000 female adult whiteflies newly emerged from virus-infected tobacco plants were collected and stored at -80°C. The RNA was extracted and sequenced using Illunima Analyzer II.
Project description:The experiment was aimed at identification of genes whose expression is up- or down-regulated in tobacco plants in response to sulfur (S) deficiency. Comparison of response to S deficit of LA Burley 21 line and AB3 line (overexpressing UP9C in antisense orientation) was done to clarify the function of LSU/UP9 family genes.
Project description:We investigated effect of severe cold to diurnal transcriptome changes in maize 3rd leaf. We used chilling-sensitive inbred line CM109. Kernels were germinated in wet sand in darkness at 25C. Seedlings were transferred to growth chamber (photoperiod 14/10h, temperature 24, light 250 umol quanta x m-2 x s-1). After full development of the third leaf (fully developed ligular region) plants were used for experiments. The experiment was begun at the start of the dark period (time zero), at which time a large sample (eight plants) was taken to serve as a reference in hybridizations. Half of the plants were transferred to cold chamber (day/night temperature 8/6°C, photoperiod 14/10 h, light 250 umol quanta•m-2•s-1) other half served as control (day/night temperature 24/22°C, other parameters was the same as for cold treatment). Further samples were taken after 200, 400, 600 (dark period) and 810, 1020, 1230, 1440 minutes (light period) of growth (total 24 hours). Each sample consisted of the middle part of the third leaf blade, pooled from three plants and frozen in liquid nitrogen. The experiment was replicated four times with two replications dye swapped.
Project description:We investigated effect of severe cold on transcriptome changes in two inbred maize lines, chilling-sensitive ETH-DL3 and chilling-tolerant ETH-DH7. Kernels were germinated in wet sand in darkness at 25C. Seedlings were transferred to growth chamber (day/night temperature 24/22C, photoperiod 14/10 h, light 250 umol quanta x m-2 x s-1) and grow in pots containing Knop's nutrient solution supplemented with Hoagland's micro-nutrients. After full development of the 3rd leaf (fully developed ligular region) plants were used in experiment. The experiment was begun at the start of the dark period. Half of the plants were transferred to cold chamber (day/night temperature 8/6C, photoperiod 14/10 h, light 250 umol quanta x m-2 x s-1) other half served as control (day/night temperature 24/22C, other parameters was the same as for cold treatment). After dark period (10h) and 200 minutes of light period samples were taken. Each sample consisted of the middle part of the 3rd leaf blade, pooled from three plants and frozen in liquid nitrogen. The experiment was replicated four times with two replications dye swapped.