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Evaluation of suitable reference genes for expression profile analyses of target genes in the coffee berry borer, Hypothenemus hampei (Ferrari) (Coleoptera: Curculionidae)

Published online by Cambridge University Press:  05 January 2024

Shaohua Zhang ,
Enhang Zhu ,
Zheng Wang ,
Yaofeng Zhong ,
Xuezong Zha ,
Hengqing Ji  and
Qianqian Meng Open the ORCID record for Qianqian Meng [Opens in a new window]
Shaohua Zhang
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 School of Plant Protection, Hainan University, Haikou 570228, 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 School of Plant Protection, Hainan University, Haikou 570228, P.R. China
Zheng Wang*
Affiliation:
Chongqing Municipal Key Laboratory for High Pathogenic Microbes, The First Batch of Key Disciplines On Public Health in Chongqing, Department of Disinfection and Vector Control, Chongqing Center for Disease Control and Prevention, Chongqing 40042, P.R. China School of Plant Protection, Hainan University, Haikou 570228, P.R. China
Yaofeng Zhong
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 School of Plant Protection, Hainan University, Haikou 570228, P.R. China
Xuezong Zha
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 School of Plant Protection, Hainan University, Haikou 570228, P.R. China
Hengqing Ji
Affiliation:
Chongqing Municipal Key Laboratory for High Pathogenic Microbes, The First Batch of Key Disciplines On Public Health in Chongqing, Department of Disinfection and Vector Control, Chongqing Center for Disease Control and Prevention, Chongqing 40042, P.R. China
Qianqian Meng*
Affiliation:
Institute of Agricultural Resources and Environment, Chongqing Academy of Agricultural Sciences, Chongqing 40042, P.R. China School of Plant Protection, Hainan University, Haikou 570228, P.R. China
*
Corresponding authors: Qianqian Meng; Email: mengzouzou@163.com; Zheng Wang; Email: sallywz618@163.com
Corresponding authors: Qianqian Meng; Email: mengzouzou@163.com; Zheng Wang; Email: sallywz618@163.com
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Abstract

The coffee berry borer, Hypothenemus hampei (Ferrari) (Coleoptera: Curculionidae), is a major destructive insect pest of coffee, which impacts the coffee crops negatively. As a draft genome has been completed for this insect, most molecular studies on gene transcriptional levels under different experimental conditions will be conducted using real-time reverse-transcription quantitative polymerase chain reactions (RT-qPCR). However, the lack of suitable internal reference genes will affect the accuracy of RT-qPCR results. In this study, the expression stability of nine candidate reference genes was evaluated under different developmental stages, temperature stress, and Beauveria bassiana infection. Data analyses were completed by four commonly used programs, BestKeeper, NormFinder, geNorm, and RefFinder. The result showed that RPL3 and EF1α combination were recommended as the most stable reference genes for developmental stages. EF1α and RPS3a combination were the top two stable reference genes for B. bassiana infection. RPS3a and RPL3 combination performed as the optimal reference genes both in temperature stress and all samples. Our results should provide a good foundation for the expression profile analyses of target genes in the future, especially for molecular studies on insect genetic development, temperature adaptability, and immune mechanism to entomogenous fungi in H. hampei.

Keywords

Beauveria bassiana infectiondevelopmental stagesexpression stabilityHypothenemus hampei (Ferrari)reference genestemperature stress
Type
Research Paper
Information
Bulletin of Entomological Research , Volume 114 , Issue 1 , February 2024 , pp. 57 - 66
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Copyright
Copyright © The Author(s), 2024. Published by Cambridge University Press

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Footnotes

*

These authors contributed equally to this study.

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