Título:	Reference genes for quantitative reverse transcription-polymerase chain reaction expression studies in wild and cultivated peanut
Autor(es):	Morgante, Carolina Vianna Guimarães, Patrícia Messenberg Martins, Andressa da Cunha Quintana Araujo, Ana Claudia Guerra de Bertioli, Soraya Cristina de Macedo Leal Bertioli, David John Brasileiro, Ana Cristina Miranda
Assunto:	Amendoim Reação em cadeia de polimerase Genes
Data de publicação:	9-Set-2011
Editora:	BioMed Central
Referência:	MORGANTE, Carolina Vianna et al. Reference genes for quantitative reverse transcription-polymerase chain reaction expression studies in wild and cultivated peanut. BMC Research Notes, v. 4, Article 339, 9 set. 2011. Disponível em: <https://bmcresnotes.biomedcentral.com/articles/10.1186/1756-0500-4-339>. Acesso em: 20 jun. 2017. doi: https://bmcresnotes.biomedcentral.com/articles/10.1186/1756-0500-4-339.
Abstract:	Background: Wild peanut species (Arachis spp.) are a rich source of new alleles for peanut improvement. Plant transcriptome analysis under specific experimental conditions helps the understanding of cellular processes related, for instance, to development, stress response, and crop yield. The validation of these studies has been generally accomplished by quantitative reverse transcription-polymerase chain reaction (qRT-PCR) which requires normalization of mRNA levels among samples. This can be achieved by comparing the expression ratio between a gene of interest and a reference gene which is constitutively expressed. Nowadays there is a lack of appropriate reference genes for both wild and cultivated Arachis. The identification of such genes would allow a consistent analysis of qRT-PCR data and speed up candidate gene validation in peanut. Results: A set of ten reference genes were analyzed in four Arachis species (A. magna; A. duranensis; A. stenosperma and A. hypogaea) subjected to biotic (root-knot nematode and leaf spot fungus) and abiotic (drought) stresses, in two distinct plant organs (roots and leaves). By the use of three programs (GeNorm, NormFinder and BestKeeper) and taking into account the entire dataset, five of these ten genes, ACT1 (actin depolymerizing factorlike protein), UBI1 (polyubiquitin), GAPDH (glyceraldehyde-3-phosphate dehydrogenase), 60S (60S ribosomal protein L10) and UBI2 (ubiquitin/ribosomal protein S27a) emerged as top reference genes, with their stability varying in eight subsets. The former three genes were the most stable across all species, organs and treatments studied. Conclusions: This first in-depth study of reference genes validation in wild Arachis species will allow the use of specific combinations of secure and stable reference genes in qRT-PCR assays. The use of these appropriate references characterized here should improve the accuracy and reliability of gene expression analysis in both wild and cultivated Arachis and contribute for the better understanding of gene expression in, for instance, stress tolerance/resistance mechanisms in plants.
Licença:	© 2011 Brasileiro et al; licensee BioMed Central Ltd. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
DOI:	https://dx.doi.org/10.1186/1756-0500-4-339
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