The Selection and Validation of Reference Genes for RT-qPCR Analysis of the Predatory Natural Enemy (Hemiptera: Anthocoridae)

Overview

Journal Insects

Specialty Biology

Date 2025 Jan 8

PMID 39769538

Authors

Chengxing Wang

Zhenjuan Yin

Yu Wang

Yan Liu

Shan Zhao

Xiaoyan Dai

Ruijuan Wang

Long Su

Hao Chen

Li Zheng

Yifan Zhai

Affiliations

Soon will be listed here.

Abstract

is a highly effective natural enemy for controlling thrips, tetranychids, aphids, and various Lepidoptera pests. Nevertheless, the molecular mechanisms underlying its interactions with host pests remain unclear. Screening for optimal reference genes is a prerequisite for using reverse transcription-quantitative polymerase chain reaction (RT-qPCR) to investigate the interrelationship. Here, ten commonly used reference genes (, , , , , , , , , and ) were selected, and their expression stability across developmental stages, tissues, temperatures, and host conditions were evaluated using , which uses multiple analytical approaches (, , the method, and ). The findings suggested that the most reliable normalization can be achieved by selecting the two reference genes for all conditions, with the optimal pairs being and for the developmental stage, and for tissue, and for the host, and and for temperature. Also, the best and least stable reference genes were chosen to compare the relative transcript levels of the in various tissues, which exhibited considerable variation. Our findings will significantly enhance the reliability of RT-qPCR and provide a foundation for further research on the expression patterns of crucial genes that are implicated in the interaction between and its host pests.

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