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Quit bugging me: Phorid fly parasitoids affect expression of an immune gene in foraging fire ant workers

Cite this dataset

King, Joanie et al. (2024). Quit bugging me: Phorid fly parasitoids affect expression of an immune gene in foraging fire ant workers [Dataset]. Dryad. https://doi.org/10.5061/dryad.9zw3r22kx

Abstract

This data contains RT-qPCR and diet measurements for quantifying gene expression of red imported fire ant, Solenopsis invicta workers in the presence and absence of the parasitoid fire ant decapitating fly, Pseudacteon curvatus, and assessing the amount of food after exposure to P. curvatus, respectively. The genes investigated were the Solenopsis invicta foraging gene (Sifor), odorant binding protein 11 (OBP-11), abaecin, defensin-2 (Def-2), cytochrome P450 4C1-like (CYP4C1-like), and hymenoptaecin (hym). Ribosomal Protein 18 (RPL18) was used as the reference gene. These genes of S. invicta workers of unknown infection status in fire ant colonies exposed to decapitating flies and control colonies were observed over a 48-h period. Two diets were measured before and after in control and treatment colonies (i.e., colonies exposed to P. curvatus).

README: Data from Quit bugging me: Phorid fly parasitoids affect expression of an immune gene in foraging fire ant workers


Summary of dataset

This Microsoft Excel spreadsheet of data contains 4 tabs. Two of the tabs: Roaches and Art, are data from the food/diet measurements (these are referred to as "diet tabs"), while the Immune gene qPCRs and Sifor, Hym, oBP-11, qPCRs contain the RT-qPCR data. See the manuscript for a description of the methods.

Description of the data and file structure

The Roaches and Art. Diet tabs contain diet measurements from treatment and control red imported fire ant, Solenopsis invicta colonies. There are 10 sub-colonies (originally five colonies split into 5 sub-colonies): B2A, B1A, R1A, F2A, F1A, F1B, F2B, R1B, B1B, B2B. Colony IDs that contain an A are treatment colonies which were exposed to decapitating flies (Pseudacteon curvatus) and colonies that contain a B are control colonies (i.e., no decapitating flies were introduced). Food source was weighed in grams with a balance scale.

The tabs in the Excel file: Immune gene qPCRs and Sifor, Hym, oBP-11, qPCRs contain the output from the RT-qPCR RT-qPCR. The instrument used was a QuantStudio™ 6 Pro System machine. The Cq were outputs by the machine. Cq AVERAGE, dCt ddCt, and Fold Change were calculated afterwards.

As with the diet tabs, an A in the sample names means treatment and B denotes control. Targets are genes of interest. The Reporter used was Syber Green. Water (molecular grade) controls were removed from the Excel tabs. There were no water samples with contamination.

Any empty cell produced by the QuantStudio? 6 Pro System was replaced with the word, "null."
All other cells that were meant to be blank were also filled with, "null."

Methods

Fire ant (S. invicta) colonies were collected and split into two sub-colonies. Containers were designed and created to serve as behavioral assay arenas. Behavioral assays were conducted in a climate-controlled laboratory, started between 13:30 and 14:20 pm, and performed over 48 hours. Each treatment colony was exposed to 10 decapitating flies (P. curvatus). Each experiment lasted 48 hours.

Food was measured in grams (g) using a scale. Data was hand-collected and then transferred to Excel where basic statistics were performed (t-tests). Fire ant workers were collected during each experiment at hours: 0, 6, 24, 30, and 48 h. They were snap-frozen on dry ice and stored at -80°C prior to RNA extraction. We assayed relative expression of Sifor, OBP11, Hym, CYP4C1-like, abaecin, and Def-2, mRNA from pooled whole body fire ant workers/foragers relative to RPL18 (an endogenous control gene) using real-time qPCR techniques on a QuantStudioTM Pro System with PowerUpTM SYBRTM Green Master Mix and manufacturer’s protocols.

Statistical analyses were conducted using paired t-tests of the DCt scores to compare differences in gene expression levels in fire ant workers at each sampling time point in the control (i.e., decapitating flies absent) and treatment (i.e., decapitating flies present) colonies. 

Funding

Texas A&M University, Urban Entomology Endowment

Texas A&M University, AgriLife Research Invasive Ant Research and Management Program

Pest Management Foundation Scholarship