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Dynamic transcriptome analysis and volatile profiling of Gossypium hirsutum in response to the cotton bollworm Helicoverpa armigera

ABSTRACT: Cotton seeds (Gossypium hirsutum cv. CCRI12) were grown in a growth chamber under 29/25°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 ± 2°C, 75 ± 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 × 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 × 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.

ORGANISM(S): Gossypium raimondii Gossypium barbadense Gossypium arboreum Gossypium hirsutum

PROVIDER: GSE62158 | GEO | 2016/01/01

SECONDARY ACCESSION(S): PRJNA263325

REPOSITORIES: GEO

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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:Lycopersicon esculentum cv. Moneymaker tomato plants were grown for 4 weeks in greenhouse conditions (16h/8h light/darkness, 27 °C). L. esculentum was randomly sorted into two groups of 16 plants each and a third reference group of 6 plants. All plants were contained individually in mesh bags. One group of 16 plants was infested with 25 female adult tomato psyllids that were 2-3 days old per plant. The other two groups of 16 and 6 plants were not infested (healthy control, and reference pool). After 3 days, the adults were removed from the infested plants to allow for egg eclosion and eventually nymphal development. In this study, leaves infested and healthy plants were harvested for RNA extraction at 0, 3, 11, and 17 days after infestation with tomato psyllids. The reference pool was harvested on day 0. Tissue (all-leaflets) from at least three plants per group was collected, flash frozen in liquid nitrogen and pooled for each time point (0, 3, 11 and 17 days after infestation) to represent treatments for before feeding (0 d), adult feeding and egg deposition (3 d), 1st and 2nd instar feeding (11 d) and 3rd to 5th instar feeding (17 d ), respectively. The whole experiment was repeated three times (three biological replicates). Plant tissues were ground using a cold mortar and pestle. Total tomato leaf RNA was extracted using the hot-phenol protocol. The RNA was precipitated, pooled, cleaned with a kit and stored at –80C. Two populations of single-stranded cDNAs will be generated from the aliquots and labeled with the cyanine Cy3 and Cy5 fluorophores. RNA isolated from the reference pool (0 d) will be pooled from each replicate and will be labeled with Cy5; this is the reference sample for each hybridization. RNA isolated from leaves infested with tomato psyllids and healthy uninfested plants from each time point will be query samples labeled with Cy3. The two samples of labeled cDNA will be simultaneously hybridized to the same microarray. Keywords: Reference design

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Dynamic transcriptome analysis and volatile profiling of Gossypium hirsutum in response to the cotton bollworm Helicoverpa armigera

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