Selection and validation of experimental condition-specific reference genes for qRT-PCR in Metopolophium dirhodum (Walker) (Hemiptera: Aphididae)

Abstract Metopolophium dirhodum (Walker) (Hemiptera: Aphididae) is one of the most common aphid pests of winter cereals. To facilitate accurate gene expression analyses with qRT-PCR assays, the expression stability of candidate reference genes under specific experimental conditions must be verified...

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Autores principales:	Xinan Li, Peipan Gong, Bingting Wang, Chao Wang, Mengyi Li, Yunhui Zhang, Xiangrui Li, Haifeng Gao, Jiansong Ju, Xun Zhu
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2020
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/0b7e0a1e8e32488b8981bd1f76ebc42c
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spelling	oai:doaj.org-article:0b7e0a1e8e32488b8981bd1f76ebc42c2021-12-02T12:42:27ZSelection and validation of experimental condition-specific reference genes for qRT-PCR in Metopolophium dirhodum (Walker) (Hemiptera: Aphididae)10.1038/s41598-020-78974-z2045-2322https://doaj.org/article/0b7e0a1e8e32488b8981bd1f76ebc42c2020-12-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-78974-zhttps://doaj.org/toc/2045-2322Abstract Metopolophium dirhodum (Walker) (Hemiptera: Aphididae) is one of the most common aphid pests of winter cereals. To facilitate accurate gene expression analyses with qRT-PCR assays, the expression stability of candidate reference genes under specific experimental conditions must be verified before they can be used to normalize target gene expression levels. In this study, 10 candidate reference genes in M. dirhodum were analyzed by qRT-PCR under various experimental conditions. Their expression stability was evaluated with delta Ct, BestKeeper, geNorm, and NormFinder methods, and the final stability ranking was determined with RefFinder. The results indicate that the most appropriate sets of internal controls were SDHB and RPL8 across geographic population; RPL8, Actin, and GAPDH across developmental stage; SDHB and NADH across body part; RPL8 and Actin across wing dimorphism and temperature; RPL4 and EF1A across starvation stress; AK and RPL4 across insecticide treatments; RPL8 and NADH across antibiotic treatments; RPL8, RPL4, Actin, and NADH across all samples. The results of this study provide useful insights for establishing a standardized qRT-PCR procedure for M. dirhodum and may be relevant for identifying appropriate reference genes for molecular analyses of related insects.Xinan LiPeipan GongBingting WangChao WangMengyi LiYunhui ZhangXiangrui LiHaifeng GaoJiansong JuXun ZhuNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 10, Iss 1, Pp 1-12 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Xinan Li Peipan Gong Bingting Wang Chao Wang Mengyi Li Yunhui Zhang Xiangrui Li Haifeng Gao Jiansong Ju Xun Zhu Selection and validation of experimental condition-specific reference genes for qRT-PCR in Metopolophium dirhodum (Walker) (Hemiptera: Aphididae)
description	Abstract Metopolophium dirhodum (Walker) (Hemiptera: Aphididae) is one of the most common aphid pests of winter cereals. To facilitate accurate gene expression analyses with qRT-PCR assays, the expression stability of candidate reference genes under specific experimental conditions must be verified before they can be used to normalize target gene expression levels. In this study, 10 candidate reference genes in M. dirhodum were analyzed by qRT-PCR under various experimental conditions. Their expression stability was evaluated with delta Ct, BestKeeper, geNorm, and NormFinder methods, and the final stability ranking was determined with RefFinder. The results indicate that the most appropriate sets of internal controls were SDHB and RPL8 across geographic population; RPL8, Actin, and GAPDH across developmental stage; SDHB and NADH across body part; RPL8 and Actin across wing dimorphism and temperature; RPL4 and EF1A across starvation stress; AK and RPL4 across insecticide treatments; RPL8 and NADH across antibiotic treatments; RPL8, RPL4, Actin, and NADH across all samples. The results of this study provide useful insights for establishing a standardized qRT-PCR procedure for M. dirhodum and may be relevant for identifying appropriate reference genes for molecular analyses of related insects.
format	article
author	Xinan Li Peipan Gong Bingting Wang Chao Wang Mengyi Li Yunhui Zhang Xiangrui Li Haifeng Gao Jiansong Ju Xun Zhu
author_facet	Xinan Li Peipan Gong Bingting Wang Chao Wang Mengyi Li Yunhui Zhang Xiangrui Li Haifeng Gao Jiansong Ju Xun Zhu
author_sort	Xinan Li
title	Selection and validation of experimental condition-specific reference genes for qRT-PCR in Metopolophium dirhodum (Walker) (Hemiptera: Aphididae)
title_short	Selection and validation of experimental condition-specific reference genes for qRT-PCR in Metopolophium dirhodum (Walker) (Hemiptera: Aphididae)
title_full	Selection and validation of experimental condition-specific reference genes for qRT-PCR in Metopolophium dirhodum (Walker) (Hemiptera: Aphididae)
title_fullStr	Selection and validation of experimental condition-specific reference genes for qRT-PCR in Metopolophium dirhodum (Walker) (Hemiptera: Aphididae)
title_full_unstemmed	Selection and validation of experimental condition-specific reference genes for qRT-PCR in Metopolophium dirhodum (Walker) (Hemiptera: Aphididae)
title_sort	selection and validation of experimental condition-specific reference genes for qrt-pcr in metopolophium dirhodum (walker) (hemiptera: aphididae)
publisher	Nature Portfolio
publishDate	2020
url	https://doaj.org/article/0b7e0a1e8e32488b8981bd1f76ebc42c
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Selection and validation of experimental condition-specific reference genes for qRT-PCR in Metopolophium dirhodum (Walker) (Hemiptera: Aphididae)

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