Cargando…

Selection and Validation of Reference Genes For qRT-PCR Analysis of Rhopalosiphum padi (Hemiptera: Aphididae)

Rhopalosiphum padi (L.) (Hemiptera: Aphididae) is an important cosmopolitan pest in cereal crops. Reference genes can significantly affect qRT-PCR results. Therefore, selecting appropriate reference genes is a key prerequisite for qRT-PCR analyses. This study was conducted to identify suitable qRT-P...

Descripción completa

Detalles Bibliográficos
Autores principales: Li, Mengyi, Li, Xinan, Wang, Chao, Li, Qiuchi, Zhu, Saige, Zhang, Yunhui, Li, Xiangrui, Yang, Fengshan, Zhu, Xun
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8079785/
https://www.ncbi.nlm.nih.gov/pubmed/33935809
http://dx.doi.org/10.3389/fphys.2021.663338
_version_ 1783685287666253824
author Li, Mengyi
Li, Xinan
Wang, Chao
Li, Qiuchi
Zhu, Saige
Zhang, Yunhui
Li, Xiangrui
Yang, Fengshan
Zhu, Xun
author_facet Li, Mengyi
Li, Xinan
Wang, Chao
Li, Qiuchi
Zhu, Saige
Zhang, Yunhui
Li, Xiangrui
Yang, Fengshan
Zhu, Xun
author_sort Li, Mengyi
collection PubMed
description Rhopalosiphum padi (L.) (Hemiptera: Aphididae) is an important cosmopolitan pest in cereal crops. Reference genes can significantly affect qRT-PCR results. Therefore, selecting appropriate reference genes is a key prerequisite for qRT-PCR analyses. This study was conducted to identify suitable qRT-PCR reference genes in R. padi. We systematically analyzed the expression profiles of 11 commonly used reference genes. The ΔCt method, the BestKeeper, NormFinder, geNorm algorithms, and the RefFinder online tool were used to evaluate the suitability of these genes under diverse experimental conditions. The data indicated that the most appropriate sets of reference genes were β-actin and GAPDH (for developmental stages), AK and TATA (for populations), RPS18 and RPL13 (for tissues), TATA and GAPDH (for wing dimorphism), EF-1α and RPS6 (for antibiotic treatments), GAPDH and β-actin (for insecticide treatments), GAPDH, TATA, RPS18 (for starvation-induced stress), TATA, RPS6, and AK (for temperatures), and TATA and GAPDH (for all conditions). Our study findings, which revealed the reference genes suitable for various experimental conditions, will facilitate the standardization of qRT-PCR programs, while also improving the accuracy of qRT-PCR analyses, with implications for future research on R. padi gene functions.
format Online
Article
Text
id pubmed-8079785
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher Frontiers Media S.A.
record_format MEDLINE/PubMed
spelling pubmed-80797852021-04-29 Selection and Validation of Reference Genes For qRT-PCR Analysis of Rhopalosiphum padi (Hemiptera: Aphididae) Li, Mengyi Li, Xinan Wang, Chao Li, Qiuchi Zhu, Saige Zhang, Yunhui Li, Xiangrui Yang, Fengshan Zhu, Xun Front Physiol Physiology Rhopalosiphum padi (L.) (Hemiptera: Aphididae) is an important cosmopolitan pest in cereal crops. Reference genes can significantly affect qRT-PCR results. Therefore, selecting appropriate reference genes is a key prerequisite for qRT-PCR analyses. This study was conducted to identify suitable qRT-PCR reference genes in R. padi. We systematically analyzed the expression profiles of 11 commonly used reference genes. The ΔCt method, the BestKeeper, NormFinder, geNorm algorithms, and the RefFinder online tool were used to evaluate the suitability of these genes under diverse experimental conditions. The data indicated that the most appropriate sets of reference genes were β-actin and GAPDH (for developmental stages), AK and TATA (for populations), RPS18 and RPL13 (for tissues), TATA and GAPDH (for wing dimorphism), EF-1α and RPS6 (for antibiotic treatments), GAPDH and β-actin (for insecticide treatments), GAPDH, TATA, RPS18 (for starvation-induced stress), TATA, RPS6, and AK (for temperatures), and TATA and GAPDH (for all conditions). Our study findings, which revealed the reference genes suitable for various experimental conditions, will facilitate the standardization of qRT-PCR programs, while also improving the accuracy of qRT-PCR analyses, with implications for future research on R. padi gene functions. Frontiers Media S.A. 2021-04-14 /pmc/articles/PMC8079785/ /pubmed/33935809 http://dx.doi.org/10.3389/fphys.2021.663338 Text en Copyright © 2021 Li, Li, Wang, Li, Zhu, Zhang, Li, Yang and Zhu. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Physiology
Li, Mengyi
Li, Xinan
Wang, Chao
Li, Qiuchi
Zhu, Saige
Zhang, Yunhui
Li, Xiangrui
Yang, Fengshan
Zhu, Xun
Selection and Validation of Reference Genes For qRT-PCR Analysis of Rhopalosiphum padi (Hemiptera: Aphididae)
title Selection and Validation of Reference Genes For qRT-PCR Analysis of Rhopalosiphum padi (Hemiptera: Aphididae)
title_full Selection and Validation of Reference Genes For qRT-PCR Analysis of Rhopalosiphum padi (Hemiptera: Aphididae)
title_fullStr Selection and Validation of Reference Genes For qRT-PCR Analysis of Rhopalosiphum padi (Hemiptera: Aphididae)
title_full_unstemmed Selection and Validation of Reference Genes For qRT-PCR Analysis of Rhopalosiphum padi (Hemiptera: Aphididae)
title_short Selection and Validation of Reference Genes For qRT-PCR Analysis of Rhopalosiphum padi (Hemiptera: Aphididae)
title_sort selection and validation of reference genes for qrt-pcr analysis of rhopalosiphum padi (hemiptera: aphididae)
topic Physiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8079785/
https://www.ncbi.nlm.nih.gov/pubmed/33935809
http://dx.doi.org/10.3389/fphys.2021.663338
work_keys_str_mv AT limengyi selectionandvalidationofreferencegenesforqrtpcranalysisofrhopalosiphumpadihemipteraaphididae
AT lixinan selectionandvalidationofreferencegenesforqrtpcranalysisofrhopalosiphumpadihemipteraaphididae
AT wangchao selectionandvalidationofreferencegenesforqrtpcranalysisofrhopalosiphumpadihemipteraaphididae
AT liqiuchi selectionandvalidationofreferencegenesforqrtpcranalysisofrhopalosiphumpadihemipteraaphididae
AT zhusaige selectionandvalidationofreferencegenesforqrtpcranalysisofrhopalosiphumpadihemipteraaphididae
AT zhangyunhui selectionandvalidationofreferencegenesforqrtpcranalysisofrhopalosiphumpadihemipteraaphididae
AT lixiangrui selectionandvalidationofreferencegenesforqrtpcranalysisofrhopalosiphumpadihemipteraaphididae
AT yangfengshan selectionandvalidationofreferencegenesforqrtpcranalysisofrhopalosiphumpadihemipteraaphididae
AT zhuxun selectionandvalidationofreferencegenesforqrtpcranalysisofrhopalosiphumpadihemipteraaphididae