Selection of reference genes suitable for normalization of qPCR data under abiotic stresses in bioenergy crop Arundo donax L.

Abstract Suitable reference gene selection in qRT-PCR is a key pre-requisite to produce reliable data in gene expression analyses. In this study, novel primers for six commonly used reference genes (AC1, TLF, Act2, TUB α, EF-1α and GAPDH) plus two new candidates (pDUF221 and RPN6) were designed and...

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Autores principales:	Michele Poli, Silvio Salvi, Mingai Li, Claudio Varotto
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/f68f5b8ea68841b59c93f3ad97482bd0
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spelling	oai:doaj.org-article:f68f5b8ea68841b59c93f3ad97482bd02021-12-02T15:05:16ZSelection of reference genes suitable for normalization of qPCR data under abiotic stresses in bioenergy crop Arundo donax L.10.1038/s41598-017-11019-02045-2322https://doaj.org/article/f68f5b8ea68841b59c93f3ad97482bd02017-09-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-11019-0https://doaj.org/toc/2045-2322Abstract Suitable reference gene selection in qRT-PCR is a key pre-requisite to produce reliable data in gene expression analyses. In this study, novel primers for six commonly used reference genes (AC1, TLF, Act2, TUB α, EF-1α and GAPDH) plus two new candidates (pDUF221 and RPN6) were designed and comparatively tested for expression stability under abiotic stresses (osmotic, heavy metal and heat shock) in shoot, root and their combination of Arundo donax L., a raising non-food energy crop. Expression stability rankings from the most to the least stable gene in each condition and in two tissues (young shoots and roots) were generated with geNorm, NormFinder and BestKeeper programs. All programs provided similar rankings and, strikingly, in most cases identified one of the new candidates, RPN6, as the most suitable reference gene. This novel set of reliable references allows to choose either the best combination of reference genes across multiple stress/organ conditions or to select condition-specific genes that can improve the quality of qRT-PCR analysis. This work provides a solid basis for the functional characterization of A. donax, by enabling accurate quantification of the transcriptional responsiveness under a series of common stress conditions of any gene of interest in this promising biomass/bioenergy species.Michele PoliSilvio SalviMingai LiClaudio VarottoNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-11 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Michele Poli Silvio Salvi Mingai Li Claudio Varotto Selection of reference genes suitable for normalization of qPCR data under abiotic stresses in bioenergy crop Arundo donax L.
description	Abstract Suitable reference gene selection in qRT-PCR is a key pre-requisite to produce reliable data in gene expression analyses. In this study, novel primers for six commonly used reference genes (AC1, TLF, Act2, TUB α, EF-1α and GAPDH) plus two new candidates (pDUF221 and RPN6) were designed and comparatively tested for expression stability under abiotic stresses (osmotic, heavy metal and heat shock) in shoot, root and their combination of Arundo donax L., a raising non-food energy crop. Expression stability rankings from the most to the least stable gene in each condition and in two tissues (young shoots and roots) were generated with geNorm, NormFinder and BestKeeper programs. All programs provided similar rankings and, strikingly, in most cases identified one of the new candidates, RPN6, as the most suitable reference gene. This novel set of reliable references allows to choose either the best combination of reference genes across multiple stress/organ conditions or to select condition-specific genes that can improve the quality of qRT-PCR analysis. This work provides a solid basis for the functional characterization of A. donax, by enabling accurate quantification of the transcriptional responsiveness under a series of common stress conditions of any gene of interest in this promising biomass/bioenergy species.
format	article
author	Michele Poli Silvio Salvi Mingai Li Claudio Varotto
author_facet	Michele Poli Silvio Salvi Mingai Li Claudio Varotto
author_sort	Michele Poli
title	Selection of reference genes suitable for normalization of qPCR data under abiotic stresses in bioenergy crop Arundo donax L.
title_short	Selection of reference genes suitable for normalization of qPCR data under abiotic stresses in bioenergy crop Arundo donax L.
title_full	Selection of reference genes suitable for normalization of qPCR data under abiotic stresses in bioenergy crop Arundo donax L.
title_fullStr	Selection of reference genes suitable for normalization of qPCR data under abiotic stresses in bioenergy crop Arundo donax L.
title_full_unstemmed	Selection of reference genes suitable for normalization of qPCR data under abiotic stresses in bioenergy crop Arundo donax L.
title_sort	selection of reference genes suitable for normalization of qpcr data under abiotic stresses in bioenergy crop arundo donax l.
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/f68f5b8ea68841b59c93f3ad97482bd0
work_keys_str_mv	AT michelepoli selectionofreferencegenessuitablefornormalizationofqpcrdataunderabioticstressesinbioenergycroparundodonaxl AT silviosalvi selectionofreferencegenessuitablefornormalizationofqpcrdataunderabioticstressesinbioenergycroparundodonaxl AT mingaili selectionofreferencegenessuitablefornormalizationofqpcrdataunderabioticstressesinbioenergycroparundodonaxl AT claudiovarotto selectionofreferencegenessuitablefornormalizationofqpcrdataunderabioticstressesinbioenergycroparundodonaxl
_version_	1718388931401089024

Selection of reference genes suitable for normalization of qPCR data under abiotic stresses in bioenergy crop Arundo donax L.

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