Unknown,Transcriptomics,Genomics,Proteomics

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Hybrid_Poplar_H11-11_Leaves_Local_Stress_Response_DataSet3

ABSTRACT: We investigated the transcriptional response of hybrid poplar (Populus trichocarpa x deltoides) leaves to a variety of stress treatments (insect feeding by Malacosoma disstria larvae, mechanical wounding, wounding plus the application of insect oral secretions, and methyl jasmonate spray) over a 24 hour time course. Experiments were conducted using clonal trees under greenhouse conditions at the University of British Columbia. We used the 15.5K poplar cDNA microarray platform previously described by Ralph et al. (Molecular Ecology 2006, 15:1275-1297). Differentially expressed genes were determined using three criteria: fold-change between treated and untreated control leaves > 1.5-fold, P value < 0.05 and Q value < 0.05. This study identified > 1,000 differentially expressed genes in response to insect feeding and/or treatments that mimic insect feeding damage. For each stress treatment, we compared RNA from leaves of four individual treated trees versus four individual control trees, with dye balance, using a total of 16 slides for the four treatments examined. Based on an earlier study using pooled biological replicates (90 trees) to examine a large combination of 18 treatments and time points, we selected the peak response time point for each treatment for further investigation using individual tree replicates. The time points and treatments examined were as follows: 24 hours for insect feeding, 6 hours for mechanical wounding, and 2 hours for both wouding plus oral secretions and methyl jasmonate. In the original experiment, leaves were harvested 2, 6 or 24 hours after the initiation of each treatment. For each tree, the lowest five mature leaves were caged in nylon mesh bags, treated or left untreated as a control. As a control for methyl jasmonate treatments, we used a tween solvent.

ORGANISM(S): Populus trichocarpa x Populus deltoides

SUBMITTER: Steven Ralph

PROVIDER: E-GEOD-14081 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Cotton seeds (Gossypium hirsutum cv. CCRI12) were grown in a growth chamber under 29/25Â°C temperature and a 16:8 h light:dark cycle, and water was added every two days. All plants were used in experiments at the 6-7 fully expanded true leaf stage, which occurred 5-6 weeks after sowing. Cotton bollworm (CBW; Helicoverpa armigera) larvae were reared on an artificial diet and maintained at 27 Â± 2Â°C, 75 Â± 10% relative humidity, and 14:10 h light:dark in the laboratory. For insect treatment, seven H. armigera larvae (third instars) were placed on a group of three plants, which were kept within plastic bags (30 Ã? 40 cm), until time of harvest, with samples for each time point maintained separately. Undamaged plants maintained under the same conditions were used as controls. Cotton leaves from control plants and plants exposed to H. armigera were harvested at 6 h, 12 h, 24 h, and 48 h after onset of herbivory. For each treatment group and time point, cotton leaves were harvested from the three plants per treatment group and flash frozen in liquid nitrogen. For each time point, three replicate treatments and controls were performed. For insect treatment, seven H. armigera larvae (third instars) were placed on a group of three plants, which were kept within plastic bags (30 Ã? 40 cm), until time of harvest, with samples for each time point maintained separately. Undamaged plants maintained under the same conditions were used as controls. Cotton leaves from control plants and plants exposed to H. armigera were harvested at 6 h, 12 h, 24 h, and 48 h after onset of herbivory. For each treatment group and time point, cotton leaves were harvested from the three plants per treatment group and flash frozen in liquid nitrogen. For each time point, three replicate treatments and controls were performed. In this study we present dynamic transcriptome analysis and volatile profiling of cotton plants fed upon by larvae of a leaf-chewing herbivore CBW. Plant transcriptomic changes induced by CBW were analyzed using Affymetrixâ??s Cotton GeneChips. Samples from a time course of six hour to 48 hours following onset of CBW feeding were analyzed to identify target genes and key pathways involved in the activation of herbivory-induced indirect defense and to explore genetic basis of such defense. In addition, we monitored the accumulation of VOCs, which represent changes in cotton plant phenotype, following CBW infestation.

Project description:Arabidopsis thaliana plants (ecotype Landsberg erecta) were grown at 20ËC constant temperature, 8 hr/16 hr Light/Dark photoperiod at 50-60% ambient humidity, for 8 to 9 weeks. Short day conditions prevented the onset of flowering and the plants were thus maintained in growth stage 1 (leaf production) with 13 to 15 rosette leaves larger than 1mm (stage 1.13 to 1.14). Diamondback moth (DBM, Plutella xylostella) larvae were maintained on cabbage (Brassica oleracea) plants in a climate-controlled room at 25ÂºC, 12 hr photoperiod with 50%-60% relative humidity. Two days before exposing A. thaliana plants to herbivore treatment, plants were transferred to a climate-controlled room (22ÂºC, 50-60% humidity, 12 hr photoperiod). For insect treatment, seven Diamondback moth (DBM, Plutella xylostella) larvae (third to fifth instars) were placed on a group of four or five plants until time of harvest, for each time point separately. All rosette leaves were harvested at 1h, 4h, 12h, and 24h after onset of continuous herbivory and flash frozen in liquid nitrogen. Control plants were maintained under the same conditions and harvested in parallel. Keywords: time course, stress response, herbivory response For each time point (1h, 4h, 12h, and 24h) two biological replicates (DBM treatment) were performed. Control plants were harvested in parallel. Total RNA from each sample was used for two labelling reactions using dye-flips (Cy5 or Cy3) and labelled cDNA from treatment and control plants were co-hybridized. Thus, for each time-point four replicates were analyzed each comparing treatment with the corresponding control (rep1 and rep2 are dye-flips of biological replicate 1, and rep3 and rep4 are dye-flips of biological replicate 2). For background correction, we defined the mean of the lowest 10% of spot intensities from a particular subgrid as the background for that subgrid. This mean was subtracted from each spot in the subgrid. Signal intensities that did not exceed the background plus 3 standard deviations thereof were defined as not detectable and were excluded from further analyses. We normalized using loess curves thus generating log2 transformed expression ratios comparing DBM treatment with control. These ratios are given in the VALUE columns of each array.

Dataset Information

Hybrid_Poplar_H11-11_Leaves_Local_Stress_Response_DataSet3

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