Reference gene selection for qRT-PCR analysis in the sweetpotato whitefly, Bemisia tabaci (Hemiptera: Aleyrodidae).

<h4>Background</h4>Accurate evaluation of gene expression requires normalization relative to the expression of reliable reference genes. Expression levels of "classical" reference genes can differ, however, across experimental conditions. Although quantitative real-time PCR (qR...

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Autores principales:	Rumei Li, Wen Xie, Shaoli Wang, Qingjun Wu, Nina Yang, Xin Yang, Huipeng Pan, Xiaomao Zhou, Lianyang Bai, Baoyun Xu, Xuguo Zhou, Youjun Zhang
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/39522e033f474e09bd2664e2fa2bc3ce
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spelling	oai:doaj.org-article:39522e033f474e09bd2664e2fa2bc3ce2021-11-18T08:02:23ZReference gene selection for qRT-PCR analysis in the sweetpotato whitefly, Bemisia tabaci (Hemiptera: Aleyrodidae).1932-620310.1371/journal.pone.0053006https://doaj.org/article/39522e033f474e09bd2664e2fa2bc3ce2013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23308130/?tool=EBIhttps://doaj.org/toc/1932-6203<h4>Background</h4>Accurate evaluation of gene expression requires normalization relative to the expression of reliable reference genes. Expression levels of "classical" reference genes can differ, however, across experimental conditions. Although quantitative real-time PCR (qRT-PCR) has been used extensively to decipher gene function in the sweetpotato whitefly Bemisia tabaci, a world-wide pest in many agricultural systems, the stability of its reference genes has rarely been validated.<h4>Results</h4>In this study, 15 candidate reference genes from B. tabaci were evaluated using two Excel-based algorithms geNorm and Normfinder under a diverse set of biotic and abiotic conditions. At least two reference genes were selected to normalize gene expressions in B. tabaci under experimental conditions. Specifically, for biotic conditions including host plant, acquisition of a plant virus, developmental stage, tissue (body region of the adult), and whitefly biotype, ribosomal protein L29 was the most stable reference gene. In contrast, the expression of elongation factor 1 alpha, peptidylprolyl isomerase A, NADH dehydrogenase, succinate dehydrogenase complex subunit A and heat shock protein 40 were consistently stable across various abiotic conditions including photoperiod, temperature, and insecticide susceptibility.<h4>Conclusion</h4>Our finding is the first step toward establishing a standardized quantitative real-time PCR procedure following the MIQE (Minimum Information for publication of Quantitative real time PCR Experiments) guideline in an agriculturally important insect pest, and provides a solid foundation for future RNA interference based functional study in B. tabaci.Rumei LiWen XieShaoli WangQingjun WuNina YangXin YangHuipeng PanXiaomao ZhouLianyang BaiBaoyun XuXuguo ZhouYoujun ZhangPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 1, p e53006 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Rumei Li Wen Xie Shaoli Wang Qingjun Wu Nina Yang Xin Yang Huipeng Pan Xiaomao Zhou Lianyang Bai Baoyun Xu Xuguo Zhou Youjun Zhang Reference gene selection for qRT-PCR analysis in the sweetpotato whitefly, Bemisia tabaci (Hemiptera: Aleyrodidae).
description	<h4>Background</h4>Accurate evaluation of gene expression requires normalization relative to the expression of reliable reference genes. Expression levels of "classical" reference genes can differ, however, across experimental conditions. Although quantitative real-time PCR (qRT-PCR) has been used extensively to decipher gene function in the sweetpotato whitefly Bemisia tabaci, a world-wide pest in many agricultural systems, the stability of its reference genes has rarely been validated.<h4>Results</h4>In this study, 15 candidate reference genes from B. tabaci were evaluated using two Excel-based algorithms geNorm and Normfinder under a diverse set of biotic and abiotic conditions. At least two reference genes were selected to normalize gene expressions in B. tabaci under experimental conditions. Specifically, for biotic conditions including host plant, acquisition of a plant virus, developmental stage, tissue (body region of the adult), and whitefly biotype, ribosomal protein L29 was the most stable reference gene. In contrast, the expression of elongation factor 1 alpha, peptidylprolyl isomerase A, NADH dehydrogenase, succinate dehydrogenase complex subunit A and heat shock protein 40 were consistently stable across various abiotic conditions including photoperiod, temperature, and insecticide susceptibility.<h4>Conclusion</h4>Our finding is the first step toward establishing a standardized quantitative real-time PCR procedure following the MIQE (Minimum Information for publication of Quantitative real time PCR Experiments) guideline in an agriculturally important insect pest, and provides a solid foundation for future RNA interference based functional study in B. tabaci.
format	article
author	Rumei Li Wen Xie Shaoli Wang Qingjun Wu Nina Yang Xin Yang Huipeng Pan Xiaomao Zhou Lianyang Bai Baoyun Xu Xuguo Zhou Youjun Zhang
author_facet	Rumei Li Wen Xie Shaoli Wang Qingjun Wu Nina Yang Xin Yang Huipeng Pan Xiaomao Zhou Lianyang Bai Baoyun Xu Xuguo Zhou Youjun Zhang
author_sort	Rumei Li
title	Reference gene selection for qRT-PCR analysis in the sweetpotato whitefly, Bemisia tabaci (Hemiptera: Aleyrodidae).
title_short	Reference gene selection for qRT-PCR analysis in the sweetpotato whitefly, Bemisia tabaci (Hemiptera: Aleyrodidae).
title_full	Reference gene selection for qRT-PCR analysis in the sweetpotato whitefly, Bemisia tabaci (Hemiptera: Aleyrodidae).
title_fullStr	Reference gene selection for qRT-PCR analysis in the sweetpotato whitefly, Bemisia tabaci (Hemiptera: Aleyrodidae).
title_full_unstemmed	Reference gene selection for qRT-PCR analysis in the sweetpotato whitefly, Bemisia tabaci (Hemiptera: Aleyrodidae).
title_sort	reference gene selection for qrt-pcr analysis in the sweetpotato whitefly, bemisia tabaci (hemiptera: aleyrodidae).
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/39522e033f474e09bd2664e2fa2bc3ce
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Reference gene selection for qRT-PCR analysis in the sweetpotato whitefly, Bemisia tabaci (Hemiptera: Aleyrodidae).

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