Case Study Using Recommended Reference Genes Actin and 18S for Reverse-Transcription Quantitative Real-Time PCR Analysis in Myzus persicae.

Myzus persicae is a globally important pest with the ability to adjust to a wide range of environmental situations, and many molecular technologies have been developed and applied to understand the biology and/or control this pest insect directly. Reverse-transcription quantitative real-time PCR (RT...

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Autores principales:	Saqib Rahman, Zhenzhen Zhao, Muhammad Umair Sial, Yanning Zhang, Hongyun Jiang
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/c75269e52cc347c982188976697a1f89
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spelling	oai:doaj.org-article:c75269e52cc347c982188976697a1f892021-12-02T20:13:38ZCase Study Using Recommended Reference Genes Actin and 18S for Reverse-Transcription Quantitative Real-Time PCR Analysis in Myzus persicae.1932-620310.1371/journal.pone.0258201https://doaj.org/article/c75269e52cc347c982188976697a1f892021-01-01T00:00:00Zhttps://doi.org/10.1371/journal.pone.0258201https://doaj.org/toc/1932-6203Myzus persicae is a globally important pest with the ability to adjust to a wide range of environmental situations, and many molecular technologies have been developed and applied to understand the biology and/or control this pest insect directly. Reverse-transcription quantitative real-time PCR (RT-qPCR) is a primary molecular technology that is used to quantify gene expression. Choosing a stable reference gene is significantly important for precisely clarifying the expression level of the target gene. Actin and 18S have been recommended as stable compounds for real-time RT-qPCR in M. persicae under the tested biotic and abiotic conditions. In this study, we checked the stability of Actin and 18S by analyzing the relative expression levels of the cytochrome 450 monooxygenase family member genes CYP6CY3 and CYP6-1, carboxylesterase gene E4 and vacuolar protein sorting gene VPS11 via RT-qPCR under various conditions. The expression levels of these four target genes were normalized using both Actin and 18S individually and the combination of these two genes. Our results confirmed that Actin and 18S can be used as reference genes to normalize the expression of target genes under insecticide treatment and starvation in M. persicae. However, at the developmental stages of M. persicae, the expression of the four tested target genes was normalized stably by Actin but not 18S, with the latter presenting a problematic change with the developmental stages. Thus, the stability of reference genes in response to diverse biotic and abiotic factors should be evaluated before each RT-qPCR experiment.Saqib RahmanZhenzhen ZhaoMuhammad Umair SialYanning ZhangHongyun JiangPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 16, Iss 10, p e0258201 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Saqib Rahman Zhenzhen Zhao Muhammad Umair Sial Yanning Zhang Hongyun Jiang Case Study Using Recommended Reference Genes Actin and 18S for Reverse-Transcription Quantitative Real-Time PCR Analysis in Myzus persicae.
description	Myzus persicae is a globally important pest with the ability to adjust to a wide range of environmental situations, and many molecular technologies have been developed and applied to understand the biology and/or control this pest insect directly. Reverse-transcription quantitative real-time PCR (RT-qPCR) is a primary molecular technology that is used to quantify gene expression. Choosing a stable reference gene is significantly important for precisely clarifying the expression level of the target gene. Actin and 18S have been recommended as stable compounds for real-time RT-qPCR in M. persicae under the tested biotic and abiotic conditions. In this study, we checked the stability of Actin and 18S by analyzing the relative expression levels of the cytochrome 450 monooxygenase family member genes CYP6CY3 and CYP6-1, carboxylesterase gene E4 and vacuolar protein sorting gene VPS11 via RT-qPCR under various conditions. The expression levels of these four target genes were normalized using both Actin and 18S individually and the combination of these two genes. Our results confirmed that Actin and 18S can be used as reference genes to normalize the expression of target genes under insecticide treatment and starvation in M. persicae. However, at the developmental stages of M. persicae, the expression of the four tested target genes was normalized stably by Actin but not 18S, with the latter presenting a problematic change with the developmental stages. Thus, the stability of reference genes in response to diverse biotic and abiotic factors should be evaluated before each RT-qPCR experiment.
format	article
author	Saqib Rahman Zhenzhen Zhao Muhammad Umair Sial Yanning Zhang Hongyun Jiang
author_facet	Saqib Rahman Zhenzhen Zhao Muhammad Umair Sial Yanning Zhang Hongyun Jiang
author_sort	Saqib Rahman
title	Case Study Using Recommended Reference Genes Actin and 18S for Reverse-Transcription Quantitative Real-Time PCR Analysis in Myzus persicae.
title_short	Case Study Using Recommended Reference Genes Actin and 18S for Reverse-Transcription Quantitative Real-Time PCR Analysis in Myzus persicae.
title_full	Case Study Using Recommended Reference Genes Actin and 18S for Reverse-Transcription Quantitative Real-Time PCR Analysis in Myzus persicae.
title_fullStr	Case Study Using Recommended Reference Genes Actin and 18S for Reverse-Transcription Quantitative Real-Time PCR Analysis in Myzus persicae.
title_full_unstemmed	Case Study Using Recommended Reference Genes Actin and 18S for Reverse-Transcription Quantitative Real-Time PCR Analysis in Myzus persicae.
title_sort	case study using recommended reference genes actin and 18s for reverse-transcription quantitative real-time pcr analysis in myzus persicae.
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/c75269e52cc347c982188976697a1f89
work_keys_str_mv	AT saqibrahman casestudyusingrecommendedreferencegenesactinand18sforreversetranscriptionquantitativerealtimepcranalysisinmyzuspersicae AT zhenzhenzhao casestudyusingrecommendedreferencegenesactinand18sforreversetranscriptionquantitativerealtimepcranalysisinmyzuspersicae AT muhammadumairsial casestudyusingrecommendedreferencegenesactinand18sforreversetranscriptionquantitativerealtimepcranalysisinmyzuspersicae AT yanningzhang casestudyusingrecommendedreferencegenesactinand18sforreversetranscriptionquantitativerealtimepcranalysisinmyzuspersicae AT hongyunjiang casestudyusingrecommendedreferencegenesactinand18sforreversetranscriptionquantitativerealtimepcranalysisinmyzuspersicae
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Case Study Using Recommended Reference Genes Actin and 18S for Reverse-Transcription Quantitative Real-Time PCR Analysis in Myzus persicae.

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