Data from: Identification and functional characterization of key ecdysteroid biosynthesis and signaling genes regulating molting and metamorphosis in Dastarcus helophoroides
Data files
Feb 25, 2026 version files 63.21 KB
-
Figure_5a_and_5b.xlsx
23.34 KB
-
Figure_6a.xlsx
12.32 KB
-
Figure_6b.xlsx
10.55 KB
-
Primer_Amplification_Efficiencies.xlsx
13.83 KB
-
README.md
3.17 KB
Abstract
Dastarcus helophoroides is an important ectoparasitoid used in the biological control of longhorn beetles. Its first-instar larvae actively seek hosts and progress through four larval instars before pupation. Throughout these developmental stages, 20-hydroxyecdysone (20E) plays a pivotal role in initiating and regulating molting and metamorphosis; however, the specific genes and underlying molecular mechanisms in D. helophoroides remain uncharacterized. In this study, comparative transcriptome analysis across larval development identified differentially expressed genes (DEGs) associated with cuticle remodeling, hormone metabolism, and stage-specific metabolic and cellular reprogramming. RT-qPCR validation revealed stage-specific expression patterns of 20E biosynthesis and signaling pathway genes, with EcR, Br-C, E93, and HR39 peaking during the larval-prepupal transition, whereas CYP18A1, E74A, E75B, HR3, and FTZ-F1 were highly expressed in early instars. RNAi targeting the eight key genes (CYP315A1, CYP314A1, EcR, USP, Br-C, E93, HR39, and FTZ-F1) severely disrupted metamorphosis and reduced adult eclosion by 47.83–100%. The developmental arrests were accompanied by stage-specific morphological abnormalities, including abnormal pigmentation, wing deformities, and malformed prepupae and pupae. This study identifies key ecdysteroid biosynthesis and signaling genes as critical regulators of D. helophoroides development, providing insights into growth regulation and artificial rearing optimization in this parasitoid biocontrol agent.
Dataset DOI: 10.5061/dryad.qrfj6q5x4
Description of the data and file structure
This dataset contains the raw experimental data underlying the manuscript
Files and variables
File: Figure_6a.xlsx
Description: (a) Effects of gene silencing on mRNA expression of 20E biosynthesis and signaling genes in fourth-instar larvae of Dastarcus helophoroides.
This sheet contains RT-qPCR Cycle Threshold (Ct) values for the RNAi target genes and the reference gene α-tubulin (internal control). Fourth-instar larvae were injected with double-stranded RNA (dsRNA), and mRNA levels were quantified 48 h post-injection.
Treatment: dsGFP (control), dsCYP314A1, dsCYP315A1, dsEcR, dsUSP, dsBr-C, dsE93, dsHR39, and dsFTZ-F1 were injected separately. Each treatment was replicated thrice.
File: Figure_6b.xlsx
Description: (b) Effect of target gene silencing on the expression level of the CYP314A1.
This sheet contains raw RT-qPCR Cycle Threshold (Ct) values for the target genes and the reference gene α-tubulin (internal control) across different developmental stages.
Treatment: dsGFP (control), dsCYP315A1, dsEcR, dsUSP, dsBr-C, dsE93, dsHR39, and dsFTZ-F1. Fourth-instar larvae of Dastarcus helophoroides were injected separately and CYP314A1 expression was measured 48 h post-injection.
File: Primer_Amplification_Efficiencies.xlsx
Description: Primer amplification efficiencies of all genes
Concentration: Relative cDNA template concentration used in the RT-qPCR standard curve. 2-fold serial dilution series expressed as powers of 2 and used to construct the standard curve for efficiency calculation.
Values:
2⁰ = undiluted cDNA (original concentration)
2⁻¹ = 1:2 dilution
2⁻² = 1:4 dilution
2⁻³ = 1:8 dilution
2⁻⁴ = 1:16 dilution
2⁻⁵ = 1:32 dilution
2⁻⁶ = 1:64 dilution
Unit: Relative dilution factor.
This sheet contains RT-qPCR Cycle Threshold (Ct) values of gene-specific primers at different template concentrations for the calculation of primer amplification efficiency.
Efficiency: represents the amplification efficiency of each primer pair, calculated from the slope of the standard curve. Efficiency values are presented in decimal format.
File: Figure_5a_and_5b.xlsx
Description: Expression profiles of 20E pathway genes during D. helophoroides development (a) Expression profiles of 20E biosynthetic genes (b) Expression profiles of 20E signaling genes.
This sheet contains RT-qPCR Cycle Threshold (Ct) values for the target genes and the reference gene α-tubulin (internal control) across different developmental stages.
Treatment: Eggs (E); 1st larval instar (L1); 2nd larval instar (L2); 3rd larval instar (L3); 4th larval instar (L4); Prepupa (PP); Pupa (P); Adult (A). R1, R2, and R3 represent three independent biological replicates.
Code/software
Any program that will open a spreadsheet, such as Excel is recommended.