Suitable reference genes for RT-qPCR analysis in Dichelops melacanthus (Hemiptera: Pentatomidae)

Area of Science:

• Entomology
• Molecular Biology
• Genetics

Background:

• Accurate gene expression quantification via reverse transcription-quantitative PCR (RT-qPCR) relies on stable reference genes for normalization.
• Dichelops melacanthus is a significant agricultural pest, yet lacks validated reference genes for molecular studies.
• Understanding gene expression is vital for developing effective pest management strategies.

Purpose of the Study:

• To identify and validate stable reference genes for RT-qPCR in Dichelops melacanthus across diverse experimental conditions.
• To provide a foundation for reliable gene expression analysis in this economically important insect pest.

Main Methods:

• Evaluated the expression stability of nine candidate reference genes (nadh, sdhb, gapdh, fau, ef1a, rpl9, ube4a, gus, rps23).
• Utilized RefFinder software integrating geNorm, NormFinder, BestKeeper, and ΔCt methods.
• Assessed stability across developmental stages, body parts, sex, starvation stress, and dsRNA exposure.

Main Results:

• Identified optimal reference gene pairs for specific conditions: ef1a/nadh (developmental stages), fau/rps23 (sex), ube4a/rps23 (body parts), rpl9/fau (starvation stress), and nadh/sdhb (dsRNA exposure).
• Expression stability varied significantly depending on the experimental variable.

Conclusions:

• The validated reference genes are essential for accurate RT-qPCR normalization in Dichelops melacanthus.
• These findings will facilitate future molecular research on this pest, contributing to improved pest management strategies.