DOI: 10.5061/dryad.qrfj6q5x4
title: Data from: Identification and functional characterization of key ecdysteroid biosynthesis and signaling genes regulating molting and metamorphosis in <em>Dastarcus helophoroides</em>

description: 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.
description: # Data from: Identification and functional characterization of key
 ecdysteroid biosynthesis and signaling genes regulating molting and
 metamorphosis in *Dastarcus helophoroides* Dataset DOI:
 [10.5061/dryad.qrfj6q5x4](https://doi.org/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.