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Selection of reference genes for quantitative real-time PCR normalization in the coffee white stem borer, Xylotrechus quadripes Chevrolat (Coleoptera: Cerambycidae)

Published online by Cambridge University Press:  09 August 2021

Qianqian Meng Open the ORCID record for Qianqian Meng [Opens in a new window] ,
Benshui Shu ,
Shiwei Sun ,
Ying Wang ,
Mei Yang ,
Enhang Zhu ,
Aiqin Liu ,
Shengfeng Gao ,
Yafeng Gou  and
Zheng Wang
Qianqian Meng
Affiliation:
Hainan Provincial Key Laboratory of Genetic Improvement and Quality Regulation for Tropical Spice and Beverage Crops, Spice and Beverage Research Institute, Chinese Academy of Tropical Agricultural Sciences, Wanning 571533, P.R. China
Benshui Shu*
Affiliation:
Guangzhou City Key Laboratory of Subtropical Fruit Trees Outbreak Control, Zhongkai University of Agriculture and Engineering, Guangzhou 510000, P.R. China
Shiwei Sun
Affiliation:
Hainan Provincial Key Laboratory of Genetic Improvement and Quality Regulation for Tropical Spice and Beverage Crops, Spice and Beverage Research Institute, Chinese Academy of Tropical Agricultural Sciences, Wanning 571533, P.R. China
Ying Wang
Affiliation:
Hainan Provincial Key Laboratory of Genetic Improvement and Quality Regulation for Tropical Spice and Beverage Crops, Spice and Beverage Research Institute, Chinese Academy of Tropical Agricultural Sciences, Wanning 571533, P.R. China College of Tropical Crops, Yunnan Agricultural University, Puer 665000, P.R. China
Mei Yang
Affiliation:
Hainan Provincial Key Laboratory of Genetic Improvement and Quality Regulation for Tropical Spice and Beverage Crops, Spice and Beverage Research Institute, Chinese Academy of Tropical Agricultural Sciences, Wanning 571533, P.R. China College of Tropical Crops, Yunnan Agricultural University, Puer 665000, P.R. China
Enhang Zhu
Affiliation:
Hainan Provincial Key Laboratory of Genetic Improvement and Quality Regulation for Tropical Spice and Beverage Crops, Spice and Beverage Research Institute, Chinese Academy of Tropical Agricultural Sciences, Wanning 571533, P.R. China College of Tropical Crops, Yunnan Agricultural University, Puer 665000, P.R. China
Aiqin Liu*
Affiliation:
Hainan Provincial Key Laboratory of Genetic Improvement and Quality Regulation for Tropical Spice and Beverage Crops, Spice and Beverage Research Institute, Chinese Academy of Tropical Agricultural Sciences, Wanning 571533, P.R. China
Shengfeng Gao
Affiliation:
Hainan Provincial Key Laboratory of Genetic Improvement and Quality Regulation for Tropical Spice and Beverage Crops, Spice and Beverage Research Institute, Chinese Academy of Tropical Agricultural Sciences, Wanning 571533, P.R. China
Yafeng Gou
Affiliation:
Hainan Provincial Key Laboratory of Genetic Improvement and Quality Regulation for Tropical Spice and Beverage Crops, Spice and Beverage Research Institute, Chinese Academy of Tropical Agricultural Sciences, Wanning 571533, P.R. China
Zheng Wang*
Affiliation:
Hainan Provincial Key Laboratory of Genetic Improvement and Quality Regulation for Tropical Spice and Beverage Crops, Spice and Beverage Research Institute, Chinese Academy of Tropical Agricultural Sciences, Wanning 571533, P.R. China
*
Author for correspondence: Zheng Wang, Email: sallywz618@163.com; Aiqin Liu, Email: laq3680@163.com; Benshui Shu, Email: shubenshui@126.com
Author for correspondence: Zheng Wang, Email: sallywz618@163.com; Aiqin Liu, Email: laq3680@163.com; Benshui Shu, Email: shubenshui@126.com
Author for correspondence: Zheng Wang, Email: sallywz618@163.com; Aiqin Liu, Email: laq3680@163.com; Benshui Shu, Email: shubenshui@126.com
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Abstract

The coffee white stem borer, Xylotrechus quadripes Chevrolat (Coleoptera: Cerambycidae), is a major destructive pest of Coffea arabica L. (Gentianales: Rubiaceae), widely planted in many Asian countries, including China. Quantitative real-time polymerase chain reaction (qRT-PCR) is a common method for quantitative analysis of gene transcription levels. To obtain accurate and reliable qRT-PCR results, it is necessary to select suitable reference genes to different experimental conditions for normalizing the target gene expression. However, the stability of the expression of reference genes in X. quadripes has rarely been studied. In this study, the expression stability of nine candidate reference genes were investigated under biotic and abiotic conditions for use in qRT-PCR's normalization. By integrating the results of four algorithms of NormFinder, BestKeeper, geNorm, and RefFinder, the optimal reference gene combinations in different experimental conditions were performed as follows: RPL10a and EIF3D were the optimal reference genes for developmental stage samples, EIF4E, RPL10a, and RPS27a for tissue samples, V-ATP and EF1α for the sex samples, EIF3D and V-ATP for temperature treatment, RPS27a and RPL10a for insecticide stress, and RPL10a, RPS27a, and EF1α for all the samples. This study will help to obtain the stable internal reference genes under biotic and abiotic conditions and lay the foundation for in-depth functional research of target genes or genomics on olfactory molecular mechanisms, temperature adaptability, and insecticide resistance in X. quadripes.

Keywords

Abiotic factorsbiotic factorsexpression stabilityqRT-PCRreference genesXylotrechus quadripes Chevrolat
Type
Research Paper
Information
Bulletin of Entomological Research , Volume 112 , Issue 2 , April 2022 , pp. 151 - 161
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Copyright
Copyright © The Author(s), 2021. Published by Cambridge University Press

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