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Identification and validation of reference genes for RT-qPCR analysis in Sclerodermus guani (Hymenoptera: Bethylidae)

Published online by Cambridge University Press:  07 October 2024

Rina Zhao ,
Xiaomeng Guo ,
Ling Meng  and
Baoping Li Open the ORCID record for Baoping Li [Opens in a new window]
Rina Zhao
Affiliation:
Department of Entomology, School of Plant Protection, Nanjing Agricultural University, Nanjing, Jiangsu Province, PR China
Xiaomeng Guo
Affiliation:
Department of Entomology, School of Plant Protection, Nanjing Agricultural University, Nanjing, Jiangsu Province, PR China Research Institute of Agricultural Sciences of Zhenjiang city, Zhenjiang, Jiangsu Province, PR China
Ling Meng
Affiliation:
Department of Entomology, School of Plant Protection, Nanjing Agricultural University, Nanjing, Jiangsu Province, PR China
Baoping Li*
Affiliation:
Department of Entomology, School of Plant Protection, Nanjing Agricultural University, Nanjing, Jiangsu Province, PR China
*
Corresponding author: Baoping Li; Email: [email protected]
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Abstract

Gene expression studies in organisms are often conducted using reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR), and the accuracy of RT-qPCR results relies on the stability of reference genes. We examined ten candidate reference genes in Sclerodermus guani, a parasitoid wasp that is a natural enemy of long-horned beetle pests in forestry, including ACT, EF1α, Hsc70, Hsp70, SRSF7, α-tubulin, RPL7A, 18S, 28S, and SOD1, regarding variable biotic and abiotic factors such as body part, life stage, hormone, diet, and temperature. Data were analysed using four dedicated algorithms (ΔCt, BestKeeper, NormFinder, and geNorm) and one comparative tool (RefFinder). Our results showed that the most stable reference genes were RPL7A and EF1α regarding the body part, SRSF7 and Hsc70 regarding the diet, RPL7A and α-tubulin regarding the hormone, SRSF7 and RPL7A regarding the life stage, and SRSF7 and α-tubulin regarding temperature. To ascertain the applicability of specific reference genes, the expression level of the target gene (ACPase) was estimated regarding the body part using the most stable reference genes, RPL7A and EF1α, and the least stable one, SOD1. The highest expression level of ACPase was observed in the abdomen, and the validity of RPL7A and EF1α was confirmed. This study provides, for the first time, an extensive list of reliable reference genes for molecular biology studies in S. guani.

Keywords

gene expressionnormalisationreference geneRT-qPCRSclerodermus guani
Type
Research Paper
Information
Bulletin of Entomological Research , Volume 114 , Issue 5 , October 2024 , pp. 613 - 621
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Copyright
Copyright © The Author(s), 2024. Published by Cambridge University Press

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