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SRX25517645: GSM8429217: LAB_40_P_rep4; Manduca sexta; RNA-Seq
2 ILLUMINA (Illumina NovaSeq 6000) runs: 9.9M spots, 985M bases, 332.2Mb downloads

External Id: GSM8429217_r1
Submitted by: Kingsolver & Willett, Biology, University of North Carolina
Study: Heat disrupts host-parasitoid interactions mediated by a viral symbiont
show Abstracthide Abstract
High temperature events can disrupt species interactions, including those among hosts, symbionts, and natural enemies. Understanding the genetic and physiological processes underlying these disruptions is a critical scientific challenge in this era of anthropogenic climate change. We explore how high temperatures disrupt the interactions among an herbivorous insect host, Manduca sexta, its insect parasitoid, Cotesia congregata, and the parasitoid's symbiotic virus. In this system, high temperatures kill developing parasitoids, but not hosts. We evaluated the physiological and transcriptomic causes of thermal mismatch in ecological interactions using parasitoid egg in vitro experiments, immunological assays, and RNAseq. We found that high temperatures disrupt the capacity of the parasitoid's symbiotic virus to immunosuppress the host insect, resulting in thermal mismatch and death of the parasitoid. At the transcriptomic level, key viral genes involved in suppressing host immune pathways showed reduced expression, driven by the virus's circular genomic structure. This work is among the first to demonstrate the genetic and physiological mechanisms by which a symbiont limits the ecological functioning of host-parasite dynamics, and provides a framework for understanding how molecular processes give rise to ecological outcomes in response to high temperature events caused by climate change. Overall design: This experiment tested the impact of a heat shock at 40C on the interactions between a host insect (Manduca sexta) and a parasitoid's symbiotic virus (Cotesia congregata bracovirus). The experiment consisted of 2 treatment types (parasitism, temperature) each with 2 levels (parasitized or control; 25C or 40C). Third instar M. sexta were either parasitized by C. congregata wasps (parasitized), or injected with sephadex beads as a control (control), then several hours later placed at either the heat shock temperature (40C) or control (25C, same as rearing temperature) for 24 hours. Insects were then returned to 25C for an additional 24 hours to mitigate acute effects of temperature stress. After this recovery period, insects were flash frozen for transcriptomics. The experiment includes a lab and field population of M. sexta to investigate population effects (indicated as LAB or FLD in the sample name). Because the lab and field populations were run at different times, some lab population individuals were run alongside the field individuals as a control (indicated by LABINFIELD in sample name)
Sample: LAB_40_P_rep4
SAMN42925752 • SRS22171655 • All experiments • All runs
Organism: Manduca sexta
Library:
Name: GSM8429217
Instrument: Illumina NovaSeq 6000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: SINGLE
Construction protocol: RNA was extracted from fat body tissue of 8-10 M. sexta larvae per treatment group for each population using the Direct-zol RNA Miniprep Kit (Zymo Research) according to the manufacturer's instructions, including a DNase I (Zymo Research) treatment step to remove genomic DNA from samples. RNA quality was assessed via Qubit 4 Fluorometer High Sensitivity RNA assay and NanoDrop Lite Plus Spectrophotometer (both Thermo Fisher Scientific). RNA libraries were constructed via QuantSeq 3' mRNA-Seq FWD Library Prep Kit for Illumina and purified via Purification Module with Magnetic Beads PCR Add-on Kit (both Lexogen) according to the manufacturer's instructions. This method generates sequences from the 3' end of mRNA transcripts, and has been demonstrated as an accurate and cost-effective alternative to whole mRNAseq methods. Library quality was assessed after the final PCR step via Qubit.
Runs: 2 runs, 9.9M spots, 985M bases, 332.2Mb
Run# of Spots# of BasesSizePublished
SRR300430634,903,040490.3M165Mb2024-08-04
SRR300430704,946,969494.7M167.2Mb2024-08-04

ID:
34499401

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