Selection of <i>Bactrocera tau</i> (Walker) Reference Genes for Quantitative Real-Time PCR

The selection of appropriate reference genes is critical for standardizing quantitative real-time reverse transcription polymerase chain reaction (RT-qPCR) data, thereby ensuring accurate and reliable results of gene expression analysis. In this study, we identified 10 candidate reference genes (enc...

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Main Authors: Yutong Zhai, Yonghao Yu, Pengfei Xu, Xianru Zeng, Xiuzhen Long, Dewei Wei, Zhan He, Xuyuan Gao
Format: Article
Language:English
Published: MDPI AG 2025-04-01
Series:Insects
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Online Access:https://www.mdpi.com/2075-4450/16/5/445
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author Yutong Zhai
Yonghao Yu
Pengfei Xu
Xianru Zeng
Xiuzhen Long
Dewei Wei
Zhan He
Xuyuan Gao
author_facet Yutong Zhai
Yonghao Yu
Pengfei Xu
Xianru Zeng
Xiuzhen Long
Dewei Wei
Zhan He
Xuyuan Gao
author_sort Yutong Zhai
collection DOAJ
description The selection of appropriate reference genes is critical for standardizing quantitative real-time reverse transcription polymerase chain reaction (RT-qPCR) data, thereby ensuring accurate and reliable results of gene expression analysis. In this study, we identified 10 candidate reference genes (encoding <i>α-tubulin</i>, <i>G6PDH</i>, <i>Rab1</i>, <i>RT</i>, <i>RPS13</i>, <i>β-tubulin</i>, <i>DPH1</i>, <i>HSP90</i>, <i>GAPDH</i>, and <i>CP</i>) and evaluated their suitability for use as reference genes in the pest insect, <i>Bactrocera tau</i>. Analysis was conducted using three software-based methods—Delta CT, NormFinder, and BestKeeper—alongside the online tool RefFinder. Expression levels of these genes were analyzed across various <i>B. tau</i> developmental stages and body parts. The overall ranking of reference gene stability scores was as follows: <i>α-tubulin</i> > <i>G6PDH</i> > <i>CP</i> > <i>β-tubulin</i> > <i>RT</i> > <i>HSP90</i> > <i>GAPDH</i> > <i>DPH1</i> > <i>RPS13</i> > <i>Rab1</i>. Ultimately, <i>α-tubulin</i> and <i>G6PDH</i> were identified as the most stable reference genes for <i>B. tau</i>.
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publishDate 2025-04-01
publisher MDPI AG
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series Insects
spelling doaj-art-e6a9cdfc0995465e8ff0a7797cba42372025-08-20T03:47:57ZengMDPI AGInsects2075-44502025-04-0116544510.3390/insects16050445Selection of <i>Bactrocera tau</i> (Walker) Reference Genes for Quantitative Real-Time PCRYutong Zhai0Yonghao Yu1Pengfei Xu2Xianru Zeng3Xiuzhen Long4Dewei Wei5Zhan He6Xuyuan Gao7Plant Protection Research Institute, Guangxi Academy of Agricultural Sciences, Nanning 530007, ChinaPlant Protection Research Institute, Guangxi Academy of Agricultural Sciences, Nanning 530007, ChinaPlant Protection Research Institute, Guangxi Academy of Agricultural Sciences, Nanning 530007, ChinaPlant Protection Research Institute, Guangxi Academy of Agricultural Sciences, Nanning 530007, ChinaPlant Protection Research Institute, Guangxi Academy of Agricultural Sciences, Nanning 530007, ChinaPlant Protection Research Institute, Guangxi Academy of Agricultural Sciences, Nanning 530007, ChinaPlant Protection Research Institute, Guangxi Academy of Agricultural Sciences, Nanning 530007, ChinaPlant Protection Research Institute, Guangxi Academy of Agricultural Sciences, Nanning 530007, ChinaThe selection of appropriate reference genes is critical for standardizing quantitative real-time reverse transcription polymerase chain reaction (RT-qPCR) data, thereby ensuring accurate and reliable results of gene expression analysis. In this study, we identified 10 candidate reference genes (encoding <i>α-tubulin</i>, <i>G6PDH</i>, <i>Rab1</i>, <i>RT</i>, <i>RPS13</i>, <i>β-tubulin</i>, <i>DPH1</i>, <i>HSP90</i>, <i>GAPDH</i>, and <i>CP</i>) and evaluated their suitability for use as reference genes in the pest insect, <i>Bactrocera tau</i>. Analysis was conducted using three software-based methods—Delta CT, NormFinder, and BestKeeper—alongside the online tool RefFinder. Expression levels of these genes were analyzed across various <i>B. tau</i> developmental stages and body parts. The overall ranking of reference gene stability scores was as follows: <i>α-tubulin</i> > <i>G6PDH</i> > <i>CP</i> > <i>β-tubulin</i> > <i>RT</i> > <i>HSP90</i> > <i>GAPDH</i> > <i>DPH1</i> > <i>RPS13</i> > <i>Rab1</i>. Ultimately, <i>α-tubulin</i> and <i>G6PDH</i> were identified as the most stable reference genes for <i>B. tau</i>.https://www.mdpi.com/2075-4450/16/5/445reference genesRT-qPCR<i>Bactrocera tau</i>different stagesdifferent body parts
spellingShingle Yutong Zhai
Yonghao Yu
Pengfei Xu
Xianru Zeng
Xiuzhen Long
Dewei Wei
Zhan He
Xuyuan Gao
Selection of <i>Bactrocera tau</i> (Walker) Reference Genes for Quantitative Real-Time PCR
Insects
reference genes
RT-qPCR
<i>Bactrocera tau</i>
different stages
different body parts
title Selection of <i>Bactrocera tau</i> (Walker) Reference Genes for Quantitative Real-Time PCR
title_full Selection of <i>Bactrocera tau</i> (Walker) Reference Genes for Quantitative Real-Time PCR
title_fullStr Selection of <i>Bactrocera tau</i> (Walker) Reference Genes for Quantitative Real-Time PCR
title_full_unstemmed Selection of <i>Bactrocera tau</i> (Walker) Reference Genes for Quantitative Real-Time PCR
title_short Selection of <i>Bactrocera tau</i> (Walker) Reference Genes for Quantitative Real-Time PCR
title_sort selection of i bactrocera tau i walker reference genes for quantitative real time pcr
topic reference genes
RT-qPCR
<i>Bactrocera tau</i>
different stages
different body parts
url https://www.mdpi.com/2075-4450/16/5/445
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