Selection of Reference Genes for RT-qPCR Analysis of Wing Dimorphism in English Grain Aphid, Sitobion avenae (Hemiptera: Aphididae)

Xiangrui Li; Xinan Li; Weiwei Yan; Brad Coates; Xuguo Zhou; Chao Wang; Gao Haifeng; Yunhui Zhang; Xun Zhu

doi:10.1093/jee/toab214

Selection of Reference Genes for RT-qPCR Analysis of Wing Dimorphism in English Grain Aphid, Sitobion avenae (Hemiptera: Aphididae)

Xiangrui Li, Xinan Li, Weiwei Yan, Brad Coates, Xuguo Zhou, Chao Wang, Gao Haifeng, Yunhui Zhang, Xun Zhu

Entomology

Research output: Contribution to journal › Article › peer-review

1 Scopus citations

Abstract

The English grain aphid, Sitobion avenae (Fabricius), exhibits classic and dramatic phenotypic plasticity in wing development. Both genetic and environmental inputs contribute to the wing polyphenism in aphids, an extreme form of phenotypic plasticity in which a single genotype produces discrete winged and wingless morphs. Validated reference genes are needed to accurately normalize temporal and spatial variation in gene expression estimates by RT-qPCR. In this research, the stability of 11 candidate reference genes selected from S. avenae transcriptomes was evaluated under an array of abiotic and biotic conditions relevant to wing development. RefFinder, a comprehensive software integrating rankings from delta Ct, BestKeeper, NormFinder, and geNorm, offered a series of reference genes for every experimental condition. Overall, helicase (HEL) and ubiquitin ribosomal protein S27A fusion protein (RpS27) are suited for most of the conditions examined in this study, although exceptions do exist. Specifically, NADH dehydrogenase (Ap-NADH) and 28S ribosomal RNA (28S) are recommended for insecticide and antibiotic treatments, while ribosomal RNA L14 (RPL14) and 18S ribosomal RNA (18S) are selected for density treatment, respectively. This study provides a suite of reference genes to investigate the wing polyphenism in S. avenae, and is important for application of RT-qPCR in future experiments of novel tactics to control aphids.

Original language	English
Pages (from-to)	313-324
Number of pages	12
Journal	Journal of Economic Entomology
Volume	115
Issue number	1
DOIs	https://doi.org/10.1093/jee/toab214
State	Published - Feb 1 2022

Bibliographical note

Funding Information:
This research was supported by the National Natural Science Foundation of China (Award number: 31772163), China Agriculture Research System of MOF and MARA (CARS-03), SINOGRAIN II (CHN-17/0019): Technological Innovation to Support Environmentally-Friendly, Food Production and Food Safety Under a Changing Climate Opportunities and Challenges for Norway-China Cooperation, Tianshan Yonug Talents (2019Q051), Research and Integrated Demonstration of Green Yielding and Quality Enhancement Technologies for Major Grain Crops in Xinjiang (2021B02002). Contributions by BSC was supported by the United States Department of Agriculture (USDA) Agricultural Research Service (ARS) (CRIS Project 5030-22000-019-00D) and the Iowa Agriculture and Home Economics Experiment Station, Ames, IA (Project 3543). Any mention of products or services is for research purposes only, and does not constitute an endorsement for use by USDA. USDA is an equal opportunity employer and provider. Authors would like to thank nonymous reviewers for their constructive criticisms and omments to improve this manuscript.

Publisher Copyright:
© 2021 The Author(s) 2021.

Keywords

Normalization
RT-qPCR
Reference gene
Sitobion avenae
Wing polyphenism

ASJC Scopus subject areas

Ecology
Insect Science

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10.1093/jee/toab214

Cite this

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title = "Selection of Reference Genes for RT-qPCR Analysis of Wing Dimorphism in English Grain Aphid, Sitobion avenae (Hemiptera: Aphididae)",

abstract = "The English grain aphid, Sitobion avenae (Fabricius), exhibits classic and dramatic phenotypic plasticity in wing development. Both genetic and environmental inputs contribute to the wing polyphenism in aphids, an extreme form of phenotypic plasticity in which a single genotype produces discrete winged and wingless morphs. Validated reference genes are needed to accurately normalize temporal and spatial variation in gene expression estimates by RT-qPCR. In this research, the stability of 11 candidate reference genes selected from S. avenae transcriptomes was evaluated under an array of abiotic and biotic conditions relevant to wing development. RefFinder, a comprehensive software integrating rankings from delta Ct, BestKeeper, NormFinder, and geNorm, offered a series of reference genes for every experimental condition. Overall, helicase (HEL) and ubiquitin ribosomal protein S27A fusion protein (RpS27) are suited for most of the conditions examined in this study, although exceptions do exist. Specifically, NADH dehydrogenase (Ap-NADH) and 28S ribosomal RNA (28S) are recommended for insecticide and antibiotic treatments, while ribosomal RNA L14 (RPL14) and 18S ribosomal RNA (18S) are selected for density treatment, respectively. This study provides a suite of reference genes to investigate the wing polyphenism in S. avenae, and is important for application of RT-qPCR in future experiments of novel tactics to control aphids.",

keywords = "Normalization, RT-qPCR, Reference gene, Sitobion avenae, Wing polyphenism",

author = "Xiangrui Li and Xinan Li and Weiwei Yan and Brad Coates and Xuguo Zhou and Chao Wang and Gao Haifeng and Yunhui Zhang and Xun Zhu",

note = "Funding Information: This research was supported by the National Natural Science Foundation of China (Award number: 31772163), China Agriculture Research System of MOF and MARA (CARS-03), SINOGRAIN II (CHN-17/0019): Technological Innovation to Support Environmentally-Friendly, Food Production and Food Safety Under a Changing Climate Opportunities and Challenges for Norway-China Cooperation, Tianshan Yonug Talents (2019Q051), Research and Integrated Demonstration of Green Yielding and Quality Enhancement Technologies for Major Grain Crops in Xinjiang (2021B02002). Contributions by BSC was supported by the United States Department of Agriculture (USDA) Agricultural Research Service (ARS) (CRIS Project 5030-22000-019-00D) and the Iowa Agriculture and Home Economics Experiment Station, Ames, IA (Project 3543). Any mention of products or services is for research purposes only, and does not constitute an endorsement for use by USDA. USDA is an equal opportunity employer and provider. Authors would like to thank nonymous reviewers for their constructive criticisms and omments to improve this manuscript. Publisher Copyright: {\textcopyright} 2021 The Author(s) 2021.",

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AU - Yan, Weiwei

AU - Coates, Brad

AU - Zhou, Xuguo

AU - Wang, Chao

AU - Haifeng, Gao

AU - Zhang, Yunhui

AU - Zhu, Xun

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N2 - The English grain aphid, Sitobion avenae (Fabricius), exhibits classic and dramatic phenotypic plasticity in wing development. Both genetic and environmental inputs contribute to the wing polyphenism in aphids, an extreme form of phenotypic plasticity in which a single genotype produces discrete winged and wingless morphs. Validated reference genes are needed to accurately normalize temporal and spatial variation in gene expression estimates by RT-qPCR. In this research, the stability of 11 candidate reference genes selected from S. avenae transcriptomes was evaluated under an array of abiotic and biotic conditions relevant to wing development. RefFinder, a comprehensive software integrating rankings from delta Ct, BestKeeper, NormFinder, and geNorm, offered a series of reference genes for every experimental condition. Overall, helicase (HEL) and ubiquitin ribosomal protein S27A fusion protein (RpS27) are suited for most of the conditions examined in this study, although exceptions do exist. Specifically, NADH dehydrogenase (Ap-NADH) and 28S ribosomal RNA (28S) are recommended for insecticide and antibiotic treatments, while ribosomal RNA L14 (RPL14) and 18S ribosomal RNA (18S) are selected for density treatment, respectively. This study provides a suite of reference genes to investigate the wing polyphenism in S. avenae, and is important for application of RT-qPCR in future experiments of novel tactics to control aphids.

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Selection of Reference Genes for RT-qPCR Analysis of Wing Dimorphism in English Grain Aphid, Sitobion avenae (Hemiptera: Aphididae)

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

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